>EPA
United Slates
Environmental Protection
Agency
Pesticides and
Toxic Sbustances
(H-7504C)
540/09-90-078
FIFRA ACCELERATED
REREGISTRATION
PHASE 3 TECHNICAL
GUIDANCE
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1 O Sr 4 ?
w UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
,1o
WASHINGTON, DC 20460
i: p o1t 0
DE 21 1989
PESTICIDES AND TOXIC SUBSTANCES
Dear Registrant:
Subject: FIFRA Accelerated Reregistration - Phase 3 Technical
Guidance
The purpose of this guidance is to provide information to
assist you in the preparation of your Phase 3 response for
reregistration of your products. You must submit a Phase 3
response for every active ingredient for which you are
responsible for providing generic data for reregistration. This
guidance document will assist you in preparing Phase 3 responses
for the studies that you consider adequate to support
reregistration of your products.
Phase 3 of reregistration is part of a five-phase process
for the accelerated reregistration of pesticides established by
Section 4 of the Federal Insecticide, Fungicide and Rodenticide
Act (FIFRA). Phase 1, now complete, involved the development and
publication of Lists A, B, C, and D of pesticide active
ingredients requiring reregistration. Phase 2, to be completed
by January 24, 1990, is the initial response by all registrants
indicating their desire to be reregistered, enumerating existing
studies that fulfill data requirements, and obligating themselves
to fill data gaps.
In Phase 3, registrants who seek reregistration but who are
not eligible for a generic data exemption must determine which of
their previously submitted scientific studies are adequate to
support reregistration. To do so, they must apply certain
criteria of data acceptability that the Agency has defined. In
addition, they must summarize each study they wish to rely on for
reregistration, reformat certain studies submitted before
January 1, 1982, in accordance with Agency guidance, and identify
information on adverse effects.
This document contains guidance required by Section 4(e) of
FIFRA concerning the adequacy of studies, how studies must be
summarized and reformatted, and how to identify information on
adverse effects. It also contains guidance on how to determine
whether to generate data on anticipated residues of pesticides on
food and feed, and an overview of EPA’S current guidance on
toxicology and residue chemistry.
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This guidance document contains all the information
necessary to begin preparing your Phase 3 response.
Supplementary guidance will be mailed to you containing specific
information relevant to your company and your Phase 2 response
after EPA has reviewed your Phase 2 response. This supplementary
guidance will contain a form that you must use to summarize your
Phase 3 response to EPA.
Phase 3 responses are due to the Agency no later than one
year after publication of Lists B, C, and D in the FEDERAL
REGISTER. Phase 3 responses for List B are due on May 25, 1990;
Phase 3 responses for List C are due on July 24, 1990; and Phase
3 responses for List D are due on October 24, 1990. Failure to
provide an adequate Phase 3 response in a timely manner may lead
to cancellation of product registration without a hearing.
This guidance document and the instructions for your Phase 3
response has been approved by the Office of Management and Budget
(0MB) and has been assigned 0MB No. 2070-0106.
If, after reviewing this document, you do not understand the
requirements for your Phase 3 response, call the Chemical Review
Manager for your chemical in the Generic Chemical Support Branch
at 1-800-552-8879 for further information. The Agency will
assist you in every reasonable way.
Caxnpt, Direg’€or
Pesticides Pfograms
Sincerel
Enclosure
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
TABLE OF CONTENTS
TABLE OF CONTENTS ....
111
LIST OF ABBREVIATIONS
v
1. HOW TO USE THIS PACKAGE l
1.1 What is the purpose of this package? '.'.'.'.'. i
1.2 What does this package contain? '.'.'.'.'.'.'.'.
1.3 How to use this package to organize your Phase 3 response '.'.'.'.'.'.'. i
1.4 When will you receive the 'Phase 3 Chemical Response Worksheet' and the 'Phase 3
Report and Worksheet on Product Status' for your active ingredients? 3
1.5 How to obtain additional information on Phase 3 submissions '.'.'.'.'.''..'. 3
2. HOW TO SUMMARIZE STUDIES 4
2.1 Acceptance Criteria 4
2.2 Traditional Laboratory Summary '.'.'.'.'.'.'.'.'.'. 4
2.3 Evidence that the Summary was Reviewed by EPA and Found Acceptable 6
2.4 Identification of Test Material 6
2.5 Guidance Specific to Individual Studies 6
2.6 Raw Data '.'.'.'.'.'.'.'.'.'.'.'.'. 7
3. REFORMATTING STUDIES 8
3 1 What data must be reformatted? \[[[ 8
3 2 How must these data be reformatted? '.'.'.'.'.'.'.'.'. 8
3.3 What information is available as guidance from NTIS on reformatting data? 8
3.4 Is any additional information needed for reformatting specific studies, other than guidance
available from NTIS? 9
3.5 How should registrants reformat data for the residue chemistry studies for which specific
data reporting guidelines are not available? 9
4. IDENTIFYING INFORMATION REGARDING UNREASONABLE ADVERSE EFFECTS 10
4.1 What Information Must be Identified? !0
4.2 How to Identify Information in Your Phase 3 Submission '.'.'.'.'.'.'.'.'.'.'.'. 10
5. GENERATING DATA ON ANTICIPATED RESIDUES 13
5.1 Degradation Studies (Storage, Transport, Processing, Cooking, Washing, Peeling) ! 13
5.2 Monitoring Data ' '' ' ' 14
APPENDDC A: SAMPLE SUMMARIES A-1
APPENDIX B: PR NOTICE 89-3 n ,
D-I
APPENDDC C: SPECIFIC GUIDANCE BY GUIDELINE REFERENCE NUMBER C-l
APPENDIX D: OVERVIEW DOCUMENT FOR SUBDIVISION F D-l
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December 24, 1989
PHASE 3 TECHNICAL GUiDANCE
APPENDIX E: OVERVIEW DOCUMENT FOR SUBDIVISION 0 E-i
APPENDiX F: INSTRUCTIONS FOR ELECTRONIC REPORTING F-i
APPENDIX G: BLANK FORMS FOR PHASE 3 SUBMISSIONS 0-i
APPENDIX H: ADDmONAL GUIDANCE ON RAW DATA H-I
APPENDIX I: GUIDELINES FOR IDENTIFYING INFORMATION CONCERNING
UNREASONABLE ADVERSE EFFECTS PURSUANT TO FEFRA SECTiON 4(e)(1)E.... I-I
LIST OF FIGURES
Figure 1. Identifying unreasonable adverse effects on the Phase 3 Chemical Response Worksheei ii
Figure 2. Identificanon of information on adverse effects that is nor indicated on the Phase 3 Chemical
Response WorksheeL Copies of this form are provided in Appendix G for your use 12
LIST OF TABLES
TABLE 1. INFORMATION TO APPEAR IN THE BODY OF A STUDY SUMMARY 5
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PHASE 3 TECHNICAL GUIDANCE
LIST OF ABBREVIATIONS AND ACRONYMS
AD! - Acceptable Daily Intake
Al. Active Ingredient
B.P. - Boiling Point
b.u.n. - blood urea nitrogen
CAS - Chemical Abstract Service
CFU. Colony Forming Units
cm - centimeters
CSPI. Center for Science in the Public Interest
DRES - Dietary Residue Evaluation System
DRG. Data Reporting Guidelines
EC25 - Effective Concentration 25
ECSO - Effective Concentration 50
EP - End-use Product
EPA. Environmental Protection Agency
FAO - Food and Agriculture Organization
FAT - Food Additive Tolerance
FDA. Food and Drug Administration
FFDCA - Federal Food, Drug and Cosmetic Act
FIFRA - Federal Insecticide, Fungicide and Rodenticide Act
GLP - Good Laboratory Practices
GPA - Gallons per Acre
HPLC- High Performance Liquid Chromatography
LUPAC. International Union of Pure and Applied Chemistry
kg. kilogram
L- Liters
LC5O - Lethal Concentration 50
LD5O- Lethal Dose 50
LOD - Limit of Detection
LSC - Liquid Scintillation Count
MATC - Maximum Allowable Toxi cant Concentration
meq - milllequivalenrs
mg. milligrams
m l , - milliliters
mm milhimeteri
MP - Manufacturing Use Product
MPCA - Microbial Pest Control Agent
MRID- Master Record Identification Number
MS - M8ss Spectroscopy
MiD - Maximum Tolerated Dose
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PHASE 3 TECHNICAL GUIDANCE
LL F OF ABBREVIATIONS AND ACRONYMS a niinuad
NFPA - National Food Processors Association
NOAEL - No Observed Adverse Effect Level
NOEL - No Observed Effect Level
NTIS - National Technical Information Service
o/oo - parts per thousand
OECD - Organization of Economic Cooperation and Development
0MB - Office of Management and Budget
OPP - Office of Pesticide Programs
PAG- Pesticide Assessment Guidelines
PAIRA - Pure Active Ingredient Radlolabelled
PFU - Plaque Forming Units
PHI - Preharvest Interval
ppm - pans per million
r.a.c. - raw agricultural commodity
SEP - Standard Evaluation Procedure
TAS - Tolerance Assessment System
TGAI - Technical Grade Active Ingredient
TLC - Thin Layer Chromatography
TRR - Total Radioactive Residue
&g. micrograms
ULV - Ultra Low Volume
USDA - United States Department of Agriculture
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PHASE 3 TECHNICAL GULDANCE
1. HOW TO USE THIS PACKAGE
1.1 What is the purpose of this p 1ckage?
Use this package to help prepare your Phase 3 submission to the Environmental Protection Agency
(EPA or the Agency). If you have completed, or will complete, Part B of the “Reregistration Phase 2
Response Worksheet for a chemical on reregistration lists B, C, and D, EPA will be mailing you the
‘Phase 3 Chemical Response Worksheet and the ‘Phase 3 Report and Worksheet on Product Status.”
These forms will be specific to your company and product(s). Please read and follow the technical
guidance in this package to evaluate enstzng studies, to szunmanze studies, and to reformat studies. Use the
Phase 3 workrheets as a final swnma,y of your response
1.2 What does this package arntain?
The package contains the following guidance for reregistration, prepared by EPA as mandated by
section 4(e) of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA):
• How to determine if a study meets EPA’s acceptance criteria (Chapter 2 and Appendix C)
• How to summarize studies (Chapter 2 and Appendix C)
• How to reformat studies (Chapter 3)
• How to identify information on unreasonable adverse effects (Chapter 4 and Appendix I).
EPA has also prepared additional guidance it believes will be useful for registrants reregistering
their products:
• A new PR Notice (PR 89-3) describing the standard format for Phase 3 summaries and
reformatted versions of studies previously submitted under FIFRA and certain provisions of the
Federal Food, Drug, and Cosmetic Act (FFDCA) (Appendix B)
• How to determine if you should generate data on anticipated residues for pesticides used on
food or feed (Chapter 5).
• Overviews of Technical Guidance for Subdivisions F and 0 (Appendices D and E)
1.3 How to e this age to organi your Phase 3 response
To develop your Phase 3 response for an active ingredient, you should review all the information
you plan to use to support reregistration of pesticide products containing that chemical. First identify
all previous communications to EPA concerning existing studies that may support reregistration.
A study is the report of a single scientific investigation, including all supporting analyses required
for logical completeness. The communications associated with a single study should include alt related
documents, supplements, and supporting analyses previously submitted to EPA that are essential to the
understanding of the scientific investigation. (See PR Notice 89.3 for additional discussion of what
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
constitutes a study.) Registrants should identify all communications associated with a single scientific
investigation that may support reregistration and consider them as a single unu to be summarized and
reformatted, even if those communications were submitted at different times to the Agency.
Once you have identified the studies that may support reregistration, you should consider whether
those studies are likely to be considered acceptable by EPA. EPA has provided acceptance criteria to
help you assess whether a stUdy is likely to be found acceptable by the Agency. These criteria are
provided as general guidance on acceptability. EPA realizes that many studies may not meet all the
acceptance criteria but may still be determined acceptable for reregistration.
u find that a study satisfies EPA’s acceptance criteria, or believe that it should be considered in
reregutering your product even though it does not satisfy all of EPA’s acceptance criteria, you must
summarize that study in your Phase 3 response by following the guidance provided in this package. The
guidance shows registrants how they may summarize and provide justification of studies that do not
meet all of EPA’s acceptance criteria. If, however, after reviewing the acceptance criteria, you decide
not to rely on an existing study to fulfill a data requirement, you must conduct a new study or take
alternative action to meet the data requirements for reregistration. The ‘Phase 3 Chemical Response
Worksheet’ will allow you to summarize for EPA your commitments to meet or to change the data
requirements you previously indicated on your ‘Phase 2 Response Worksheet.’
In summarizing each study, you must also determine whether it contains information on
unreasonable adverse effects. Follow the guidance in Chapter 4 and Appendix I to determine the
information that must be identified in a summary and use the ‘Phase 3 Chemical Response Worksheet,’
as explained in Chapter 4, to indicate which summaries contain such information. Chapter 4 also
indicates how to identify information on unreasonable adverse effects if such information is not
associated with a study being summarized in Phase 3.
After you have identified the studies you wish to summarize, you must reformat certain residue
chemistry and toxicology studies conducted before 1982. Chapter 3 identifies which studies must be
reformatted and explains how to reformat them.
This guidance package also contains a chapter on anticipated residues (pesticide residues in food at
the time of consumption, as opposed to the tolerance, the maximum legal residue). Registrants are
encouraged to conduct such studies early in the reregistration process so that more realistic dietary
exposure assessments can be obtained for estimating dietary risks from pesticides. EPA will require
anticipated residue studies in cases in which unacceptable risks are calculated based on tolerance level
residues on food or based on tolerance level residues in conjunction with information on the percent of
crop treated with a pesticide. Chapter 5 discusses the factors registrants should consider when designing
monitoring studies and degradation studies (e.g., storage. cooking, peeling). If you decide to conduct a
study on anticipated residues, indicate your intention on the ‘Phase 3 Chemical Response Worksheet’
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PHASE 3 TECHNICAL GUIDANCE
1.4 When will rec ve the ‘Pb e 3 Qi mka1 R poi e Wor heet’ and the ?hase 3 Report and
Worksheet on Product Stanm’ r your active Ingredients?
After EPA has received and processed your ‘Phase 2 Response Worksheet,’ the Agency will send
you forms specially prepared for your active ingredients and products. You must complete the ‘Phase 3
Chemical Response Worksheer and use it as a transmittal document that summanzes the information in
your Phase 3 response.
1.5 How to obtain additional inthrmatjon on Pb e 3 submisajolLi.
For further information on the requirements of Phase 3 contact the Chemical Review Manager for
the chemical in the Generic Chemical Support Branch at (800) 552-8879.
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PHASE 3 TECHNICAL GUIDANCE
2. HOW 1D SUMMARIZE SI1.JDIES
The required elements for Phase 3 study summaries are listed in Appendix B (PR Notice 89-3), Part
VIII. Table 1 lists information that must appear in the Body of the Sum narj for two categories of
studies: (1) studies that were initiated after October 1984 and were approved by EPA, and (2) all other
studies being relied on for reregistration. An abbreviated summary may be submitted for studies in
category (1). A standard summary must be submitted for all other studies. Table 1 also indicates where
additional guidance may be found for summarizing studies. The remainder of this chapter provides
general guidance on topies relevant to study summaries and describes how to use the specific guidance
given in Appendix C. The chapter also includes a definition of raw data. As described in Appendix B
(PR Notice 89-3), registrants must certify access to raw data as part of each Phase 3 summary. Several
sample Phase 3 summaries appear in Appendix A.
2.1 Acceptance Criteria
Appendix C lists, by guideline reference number, the criteria that generally distinguish an acceptable
study from a study that cannot be used to support reregistration. EPA has provided these criteria so
registrants can assess whether a study is likely to be found acceptable by the Agency. EPA realizes that
research on pesticides is complex and does not intend the criteria to be used as an inflexible checklist.
The Agency acknowledges that acceptable studies exist that do not meet all the acceptance criteria, and
also that studies may be fundamentally flawed even though they seem to meet all the criteria. If you
believe that a study that deviates from EPA’s acceptance criteria should still be considered to support
reregistration, you may summarize that study in your Phase 3 Response.
Appendix C lists two kinds of acceptance criteria: critena that have been reviewed by EPA’s
Scientific Advisory Panel and have received public notice and comment; and supplemental criteria
(marked with an asterisk) that identify additional factors the Agency believes are useful to consider
when determining the adequacy of a study.
EPA requires the submission of acceptance criteria worksheets when registrants choose the
abbreviated summary format for studies initiated after October 1984 that have been reviewed and
approved by EPA. The submission of acceptance criteria worksheets is strongly recommended for all
summaries. A standard summary must identify any acceptance criteria not met and explain why the
study should nevertheless be considered irrespective of whether or not the acceptance criteria worksheet
is included.
2.2 T Idoullabcqizo,y Summary
If a study eftglbk for the abbreviated summary format, registrants may include in their Phase 3
Response a traditional laboratory summary that includes, as a minimum: an identification of test
materials and methods, a summary of the results of the study, and the date of test initiation and
termination. Registrants may provide a legible photocopy of the summary of the initial study report if
that summary includes the information listed above. Registrants summarizing residue chemistry studies
are also asked to identi1 the tolerance petition numbers associated with those studies, if the studies
have been used to support a tolerance.
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PHASE 3 TECHNICAl GUIDANCE
TABLE! INFORMATION TO APPEAR IN THE BODY OF A STUDY SUMMARY
ABBREVIATED SUMMARY - OPTIONAL FOR STUDIES
IN ATEI) AFFER 1984 ThAT HAVE BEEN APPROVED BY EPA
Item Guidance
Completed acceptance criteria worksheet Chapter 2.1 and Appendix C
Traditional laboratory summary Chapter 2.2
Evidence that the study was reviewed by EPA Chapter 2.3
and found acceptable
Identification of information concerning Chapter 4
unreasonable adverse effects
STANDARD SUMMARY - REQUIRED FOR ALL OThER STUDIES
Item Guidance
Completed acceptance criteria worksheet Chapter 2.1 and Appendix C
(optional)
Identification of any acceptance criteria not met Chapter 2.1
and an explanation of why the study should
nevertheless be considered
Identification of test material Chapter 2.4
Items specific to individual guideline reference Chapter 2.5 and Appendix C
numbers
Identification of information concerning Chapter 4
unreasonable adverse effects
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
2.3 Evidence thet the Swnniaiy w Reviewnd by EPA and Found Aixeptabic
If a registrant finds that a study is eligible for the abbreviated summary format, the registrant must
provide EPA with the supporting evidence that the study was reviewed by EPA and found acceptable.
This information may include a copy of EPA’s review of the study, a letter from a Product Manager
stating that EPA has found the study acceptable, or other information that the registrant typically relies
on to determine whether a study was found acceptable by EPA.
2.4 Identification of Test Material
The identification of the test material for all non-microbial pesticides should include:
• Chemical name [ Chemical Abstract Service (CAS) name preferred], common name, and trade
name. International Union of Pure and Applied Chemistry (IUPAC) designation is requested.
• CAS number of active ingredient
• Chemical structure of active ingredient
• Percent active ingredient
• Identity and composition of impunties
Unless otherwise indicated in Appendix C, EPA e rpects registrants to have used the technical grade of
the active ingredient in product chemistry studies and other studies.
If the test substance is a microbial, the identification must include:
• Scientific name and, to the extent possible, serotype and strain or other appropriate designated
type, and, to the extent possible, a qualitative and quantitative determination of composition
(including names and quantities of known contaminants and impurities, within technically
feasible limits). The determination shall also include quantities of unknown materials, if any, to
account for 100% of the sample;
• Manufacturer and lot number of the test substance, and relevant properties of the substance
tested, (i.e., physical state, pH, stability, and punty); and
• Identification and composition of any vehicles or other materials (e.g., diluents, suspending
agents, emulsiflers, virulence enhancers) used in administering the test.
2.5 Guidance sdflc to Indi v Idual Studies
Appendix C also identifies by individual Guideline Reference Number the specific information that
should be included in a summary. This guidance highlights the information of greatest importance to
EPA’s review and is designed to expedite both the registrants’ preparation of study summaries and
EPA’s review.
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PHASE 3 TECHNICAL GUIDANCE
EPA recogni c that even though a study may meet the acceptance criteria for a guideline
requirement, it may not necessarily contain all the information listed in the guidance for summarizing
studies. In that case, the summary must cojitain all the information available that addresses the specific
summary guidance provided.
If the summary guidance for a specific study calls for a ‘summary of conclusions,’ limit your
summary to one or two sentences that state the conclusions. For example, if you are summarizing a
hydrolysis study, a sentence or two indicating whether the chemical is stable in water or breaks down
rapidly is all that is needed. In contrast, the discussion of results should provide additional information
regarding the results. This should include such information as a discussion of any problems that
occurred during the study and deviation from protocols.
EPA has specified in Appendix C (for example, Guideline Reference No. 171.3) when summaries
are not required. If no summary guidance is provided for a specific guideline number and no specific
exemption or exclusion is identified, you should include in the summary an identification of test
materials and methods, results, discussion of results, conclusions, and the date of test initiation and
termination.
Z6 Raw Data
FIFRA states that each registrant seeking reregistration must submit ‘a certification that the
registrant or the EPA Administrator possesses or has access to the raw data used in or generated by’
the study that is being summarized. A standard format for this certification appears in Appendix B (PR
Notice 89.3). A certification statement must be included in each study summary.
For toxicology studies initiated or conducted after December 29, 1983, and for studies in chemistry,
environmental fate, or ecological effects, the definition of ‘raw data’ provided by the Good Laboratory
Practice (GLP) Standard serves as a guide (40 CFR 160.3). The GLP defines ‘raw data’ as
any labora:oi’y worksheer records, memoranda, notes, or exact copies thereof; that are the results of
original observations and activities of a sszuty and are necessary for the reconstruction and evaluation of
the report of that study In the event that exact Owucnpts of raw data have been prepared (e g., tapes
which have been transcribed vabanrn, date4 and verified accurate by signature) the exact copy or exact
transcript may be substituted for the original source as raw data. ‘Raw data’ may include photographs,
microfilm, or microfiche copies cwnpute pruuours magnetic media, incb ding dictated obser.’anons, and
recorded data from automated ijisn’ume,us.
EPA has described In Appendix H the specific types of information it considers to be ‘raw data’ for
studies in different iubdMslons. EPA will also consider the raw data requirement satisfied if the study
has been audited by EPA and the audit found that all raw data needed for reconstructing the study were
present.
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PHASE 3 TECHNICAL GUIDANCE
3. REFORMArFINo STIJDIES
3.1 What data must be reformatted?
FIFRA requires registrants who submitted studies concerning chronic dosing, oncogenicity,
reproductive effects, mutagenicity, acute and subchronic neurotoxicity, teratogenicity or residue chemistry
before January 1, 1982, and who are relying on those studies to support reregistration to reformat the
data from each of those studies. In addition, if the toxicology studies listed above are not required by
regulations issued under section 3 for an active ingredient, FIFRA states that registrants must reformat
the reports of acute and subchronic dosing they submitted before January 1, 1982.
EPA has decided that two categories of studies identified by FIFRA do not require special
reformatting: (1) oncogenicity studies in the mouse that have been through the Office of Pesticide
Program’s peer review and have been classified by EPA, and (2) acute toxicity studies. For those
categories of studies, the summary information registrants will supply in Phase 3 will provide sufficient
reformatted information for EPA’s review.
3.2 How must these data be reformatted?
Registrants must:
1. reformat all data developed for a single toxicology study or for a single residue chemistry study
in one report. If any report of a study was submitted before January 1, 1982, all data for that
study, even if submitted after 1982 must be included in the reformatted report. PR Notice 89-3
in Appendix B provides a definition of a study;
2. follow the standard guidance in PR Notice 89-3 concerning format standards; and
3. follow the special instructions for data reporting developed by EPA for each guideline. This
guidance is available, on request, from the National Technical Information Service (NTIS).
3.3 What information available us gnithln from NTIS on reformatting data?
EPA has made available through NTIS all the guidance relevant to reporting the data from
toxicology and residue chemistry studies for chemical pesticides. EPA has also developed an overview of
the technical guidance relevant to toxicology and residue chemistry studies for chemical pesticides. This
guidance appears In Appendices D and E. It explains the relationship between EPA’s data reporting
guidelines and the other technical guidance documents available from NTIS. This guidance is entitled
‘Overview of Su1d fon F - Ha rd Evaluation; Humans and Domestic Mimals and Overview of
Subdivision 0 -Residue Chemistry.’
Each overview lists EPA publications pertaining to the relevant subdivision and identifies how they
are related to specific guideline reference numbers. The overview also identifies EPA’s most current
guidance for generating and reporting data, clarifies some apparent inconsistencies between the Pesticide
Assessment Guidelines, Data Reporting Guidelines, and Standard Evaluation Procedures, and indicates
future directions that the Agency is considering.
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PHASE 3 TECHNICAL GU [ DMICE
Registrants seeking to reregister microbial pesticide products should consult ‘Pesticide Assessment
Guidelines, Subdivision M; Part A, Microbial Pest Control Agents’ (NTIS Number PB89-21 1676) for
guidance on reformatting toxicology and residue chemistry studies for such pesticides. Registrants who
wish to reregister biochemical pesticide products should consult ‘Pesticide Assessment Guidelines,
Subdivision M; Bioratfonal Pesticides’ (NTIS number PB83-153965) for the most current guidance on
data submission.
3.4 Is any additional information needed for reformatting sp iflc studies, other than guidance available
from NTIS?
The documents available from NTIS provide all the guidance needed for reformatting toxicology
studies. For residue chemistiy, however, specific data reporting guidelines are only available for the
major types of residue chemistry studies.
3$ How should registrants reformat data for the residue chemistry studies for which spedfic data
reporting guidelines are not available?
EPA suggests that registrants use the guidance available for reporting the study ‘Magnitude of the
Residue: Crop Field Trials’ (Guideline Reference Number: 171-4(k)J for reporting results from studies
of the ‘Magnitude of the Residue’ for potable water, fish, irrigated crops, and food handling [ Guideline
Reference Numbers: 171-4(f); 171-4(g); 171-4(h); and 171-4(J)J. Registrants should use the guidance
available from NTIS, and interpret the water, fish, irrigated crops and food being tested as the ‘Test
Commodity.’ EPA suggests that registrants use their judgement to decide when test parameters
described for the major study are not applicable (e.g., to decide that identifying soil characteristics may
not be relevant if the study concerns food handling) or need to be interpreted in a different context
(e.g., to decide that a description of a field tnal design would not be relevant to the context of a study
of a food handling use, but that a description of the restaurant where the food handling occurs would
be).
The other restd ue chemistry data requirement for which no published data reporting guidance exists
is ‘Reduction of the Residue’ (Guideline Reference Number: 171.5). The Agency suggests the use of
guidance developed for ‘Processed Food/Feed Study” [ Guideline Reference Numbec 171-4(l)J for
reporting data for this study. Where the guidance calls for information on ‘Test Procedures,’ EPA
suggests that the submitter describe how the commodity was washed, cooked or stored, prior to residue
analysis.
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PHASE 3 TECHNICAL GUIDANCE
4. JDE?rflyyij cj INFORMATION REGARDING UNREASONABLE ADVERSE UFECrS
As part of your Phase 3 submission you must identify information relevant to EPA’s determination
of whether a pesticide poses Unreasonable adverse effects. (This requirement is stated in FIFRA Section
4(e), which refers to information that must be submitted under FIFRA Section 6(a)(2).) Note that the
information need not, by itself, demonstrate unreasonable adverse effects. The information need only be
relevant to EPA’s determination of whether any adverse effects are reasonable or not.
This chapter has two parts. Section 4.1 describes how to determine which “pieces’ of information
should be identified. Section 4.2 describes how to identify the information in your Phase 3 submission.
4.1 What Information Must be Identified?
The critena described In Appendix I must be used to determine which pieces of information must
be identified. A “piece” of information is any study, incident, or other information pertaining to the
active ingredient which is known to the registrant, whether or not the information has already been
submitted to EPA, and whether or not the registrant possesses documents related to the information.
Make a list of all pieces of information. For each piece of information on the list use the guidance
in Appendix I to deterinme whether the information would be submissable under FIFRA Section
6(a)(2), assuming that the information has not yet been submitted, but all other pieces of information
on your list have been submitted to EPA.
4.2 How to Identify Information In Your Phase 3 SubmLsslon
If any piece of information would be submissable as descnbed above, it must be identified in one of
two ways:
(1). If the piece of information is part of a study that you are using in support of reregistration,
identify the information in the study summary and check ‘Y ’ in column 8 of the Phase 3
Chemical Response Worksheet, which you will receive later in a separate mailing. An example
showing how to use the worksheet to identify unreasonable adverse effects appears in Figure 1.
(2). If the piece of information is not part of a study that you are using in support of reregistration,
complete the form shown in Figure 2. Two copies of this form (“Phase 3 Identification of
Unreasonable Adverse Effects Information Not Indicated on the Phase 3 Worksheet’) are
provided in Appendix 0 for your use.
10
-------
N
December 24, 1989
PHASE 3 TECHNICAL GUIDM4CE
\\
Figure 1. Idennf zng unreasonable adverse effects on the Phase 3 Chemical Response WorksheeL
UNITED STATES INVIIONMU4TA PIOTECIION AGENCY
WASII IPICTON . D C iSO
S EPA REREGISTRATION PHASE 3 CHEMICAL RESPONSE
WORKSHEET
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11
-------
December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
Urnt.d kit., Ea ,Iroa.saaaI Psiscit.. Ag. .. ,
W—½ngt. .. D.C. WNO
PHASE 3 lD TlFICkTIoN OF
UNREASONABLE ADVERSE INFORMATION
NOT INDICATED ON THE PHASE 3 WORKSHEET
A
° C’°’
A .,at s 4w ta.as
Isuftuc cma Please type or print in ink. Please read carefully the aI ct d u uc oea and supp ly the
infurmaaon r uested on this form. Use dditional sheets(s) if i asary
FTLL OL7TTl s FORM FOR EV Y $ ARATE ITE4OP ADVD.SE U7ECTS Th IFORMATTON.
CASE
COMPANY NUMBER
NAME CHEMICAL
NUMBER
COMPANY CHEMICAL
NUMBER NAME
1. esth th ..pastu to anyso sfyourpswh Ye. No
Uye List the product numb (s)
2. lath, .our of luiwmatlon a study act ideudfled ou w Phase 3 Otsailcel Response Weelsaheet. an
boddent repast other inioona ou iowan?
U the source Is a study not ou your Wo lcshest, fill out
soc ou 3 below.
lithe source Is an thr4d .!t.r ,poet ucoth source.
fill out u oa 4 bql w.
3. A. Has the study bean submittud to D’A?
Yes h ID 4imsb
Oats Submltt,d__________
This
No Uao youu ,lequ)ndtosubuift
acyoiUwstuywtd
section Ma)(2) of FITRA.
I. Attede su asy . of f ,
toffee atlcsc out aed In 11w study sc
explain why It may bidfr i edve,.
4. A. Has the bean submitted to
Yes Da is Submitted___________
How and towti om
Subu i1 d____________
No D lf oueqo1ndiosubaIt
aaspyc1thsstedyu
section 6(a)421 of FURA.
B. cit summaz ,r of ado,,.. effects Infasmatlon
1. Identit Ies th. source Cf Inhemation (e.g. legal
— lefle. bum - 1w y repasU.
2. Desalbes the . 4ve.i. effects sod seplabs why
ltmaybiiII a.Cf,e. . s .
S. Cistifksti , .
I tSat the slatsumeis that I ha,. made ., this I.e. sad all
w. t_ _ 1 th.iela as, Iiv, .uunt. sad cumplet.. I ac1wowI.dg. that
as, biul 4ly 1 .1 ..., mwsadlag staaimast may be puatsitabi. by I
ii bath ..4,, appllc.bI. law.
S A. Sipasur, . 1 Cs .paay. Avshsoised R.pm.u*atlve
6. Canted (Typed Name)
p . .
Figure 2. Idennficanon of information on adverse effects that is not indicated on the Phase 3 Chemical
Response Worksheet. Copies of this Jo int are provided in Appendix G for your us
12
-------
December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
5. GENERATING DATA ON ANTICWATED RESIDUES
During the reregistration process mandated by FWRA, EPA will assess human health risks from
pesticide residues in food. EPA strongly encourages registrants to submit data on anticipated residues
to improve the assessment. If data are not provided voluntarily, EPA will base the assessment on less
realistic methods such as tolerances or tolerances combined with data on percent crop treated. These
estimates will be based on currently available field trial and processing data. If the risks calculated by
EPA are unacceptable, EPA will require anticipated residue data to be provided. Registrants would
then have to provide this data or propose to modify the use of their chemical by changing, for example,
the rate, number or type of applications, or by deleting uses.
Two types of data are requested -- degradation studies and monitoring data. Each type is discussed
in more detail below. Additional guidance will be available in mid-1990. Generation of these types of
data, which are not routinely required by EPA, would be beneficial to both the Agency and the
registrant since it would permit more realistic estimates of exposure that are likely to be lower than
estimates based on currently available data.
The human health risk from exposure to pesticide residues in food is a function of the toxicity of
the pesticide, the amount of food consumed, and the level of residues in the food. EPA has developed
a computer-based system called the Dietary Risk Evaluation System (DRES), formerly called the
Tolerance Assessment System (TAS), to integrate this information. DRES estimates dietary exposure in
terms of quantity of pesticide consumed per unit body weight per day and compares it to a previously
determined Acceptable Daily Intake (ADI) or similar toxicological threshold. Consumption patterns are
derived from a 1977-78 U. S. Department of Agriculture survey involving 3-day dietary records for
30,770 individuals and 3,734 food items (food forms). DRES can handle separate residue estimates for
multiple food forms of a single commodity. For example, food forms for apples include fresh apples,
cooked apple products and apple juice. Dietary exposure can be estimated for the entire U.S.
population and for 22 subgroups, determined by such factors as age, sex, ethnic background, and
geographic region.
When alternative data are not available, EPA enters tolerance level residues into DRES. This
approach will tend to overestimate exposure because a tolerance level is the ma umum allowable residue
on a commodity and should not commonly be reached or exceeded from proper product use. DRES is
also capable of using ‘anticipated residues’ which are more realistic estimates of residues in food at time
of consumption and consequently lead to more realistic estimates of exposure and risk.
The method nsed to quantify the anticipated residue depends on the toxic effect of the pesticide.
For acute effe , the most appropriate quantity might be the maximum amount of pesticide residue
likely to be consumed in a given period (e.g., meal, day, week, etc). For chronic effects, the average
residue level a longer period of time might be more appropriate.
5.1 l)egradatlon Studies (Storage, Tramport, Pro sing , Cooking, Washing Peeling)
Degradation refers to the change in the amount and composition of the total residue from harvest
to the point of consumption. For pesticides applied after harvest, degradation describes the change from
pesticide application to consumption. Two general degradation scenarios are possible. The pesticide may
13
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
degrade to form less toxic products, or the pesticide may degrade to form more toxic chemical
compounds.
The Agency does not recommend a specific protocol, but urges the registrant to submit a protocol
for Agency review before initiating a degradation study. Degradation studies for representative
commodities within a commodity group may be sufficient if commercial and home practices are similar
for the different commodities. EPA is working with the National Food Processors Association (NFPA)
to develop protocols for processing studies for some commodities. These are available from NFPA or
from the Dietaiy Exposure Branch of EPA.
After harvest, commodities can be stored (sometimes for extended periods of time), transported,
commercially processed, waxed, washed, peeled, cooked, and treated in other commodity-specific ways
(e.g. removal of the outer leaves from lettuce). A typical degradation study should take a large treated
sample through all the processes from harvest to consumption, and should simulate typical commercial
and home practices as closely as possible. Subsamples should be removed at each important point for
residue determination in the edible portion of the commodity. In all cases, but particularly when
degradation products are more toxic than the parent, application rates should be chosen which are close
to the maximum registered rates so that metabolite ratios likely to result from typical applications can
be determined. Residues in the raw commodities at the beginning of the study should be well above the
limit of detection (LOD) of the analytical method so that a decline in residues can be measured.
Analytical methods must have sufficiently low LODs so that, when calculated at the LOD, the risk from
the measured residues can be determined to be acceptable.
Time and temperature considerations are important when examining storage, transportation,
commercial processing, and cooking. Humidity may be important when examining storage and
transportation. The point at which wax is applied to some commodities must be considered (e.g. citrus,
cucumbers.) The typical way(s) commodities are washed, peeled and cooked (e.g. boiled, fried, roasted,
etc.) are important considerations. Other processes may also be important for specific commodities (e.g.
shelling nuts, removal of the outer leaves from lettuce and cabbage, removal of the thick part of the
stem from broccoli and asparagus.)
5.2 Monitoring Data
A monitoring program (or survey) is a study in which samples of agricultural commodities/foods are
obtained (usually at the retail level) and analyzed for pesticide residues. Residue levels determined in
this way are likely to be more realistic estimates of dietary residues than tolerance levels. Sampling
locations and numbers of samples obtained are chosen so that the residue levels measured are
representative of residue levels found nationally in a specific commodity and satisfy pre-determined
requirements of a racy and precision.
EPA does am rcammend any specific protocol, but urges registrants to submit the protocol for
Agency review beftwe Initiating a study. Factors to be considered when preparing a protocol include
which food forms to monitor and the point in the life of the commodity to obtain the samples, the
required accuracy and precision of the results (which influences the number of samples required), the
toxicological effect, and available analytical methodology. The following must be addressed:
food forms to be analyzed (for guidance see ‘Tolerance Assessment Syuem Food-Form Code?,
1O, 4, USEPA/OPP)
14
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
• movement of the commodity in commerce (including storage times and conditions)
• the time of year or season in which the pesticide is applied (data should be obtained at various
times or seasons throughout the year)
• the variability in growing conditions or agricultural practices for a commodity
• the percent of livestock treated
• the percent of crop treated and specific geographical areas likely to be treated based on usage
data or on the geographical range of the pest being controlled
• the percentage each specific crop coninbutes to the total dietary exposure to a pesticide (in
most cases it may be appropriate to generate data for representative commodities for crop
groupings as defined in 40 CFR 180.34(f)(9))
• whether normal use of the pesticide may have temporarily declined due to adverse publicity
(monitoring data may not be acceptable in this situation) or due to declining pest pressure
• the performance of the analytical method (if residues at the limit of detection of the analytical
method are of toxicological concern, an alternative method may be necessary)
• the appropriateness of compositing samples (taking into account toxic effects and expected
vanability of residue levels) to reduce the total number of samples required
• commodities likely to contain detectable residues as determined by field trial data
• food forms consumed by population subgroups determined to be primarily at risk
• the active ingredient
• the availability of livestock feeding studies from which residues in animal products might be
derived from residues measured in treated animal feeds
• the availability of processing data from which residues in processed commodities can be derived
from residues measured in the raw commodity
15
-------
Phase 3 Summary of MRID 888888;
Metabolism of “HERBICIDE” in Soybeans: Project 23489
Guideline Reference:
171—4(a) Nature of the Residue: Plant Study
Summary Prepared By:
A. Chemist
November 16, 1980
Original Study Prepared By:
XYZ Laboratory
Chicago, IL
Page 1 of 7
-------
Statement of Data Confidentiality Claims
Information claimed confidential on the basis of its falling within
the scope of FIFRA §lO(d)(l)(A), (B), or (C) has been removed to
a confidential appendix, and is cited by cross-reference number in
the body of the study.
Company:
Company Agent:
Title:
(Typed Name)
(Typed Name)
Signature:,
Date:
Page 2 of 7
-------
S - IPLE SU%PJARY FOR RESIDUE CT- lEWIS TRY STUDIES
Good L aboratory Practice Statement
This study does not meet the requirements of 40 CFR Part 160, and
differs in the following ways:
1.
2.
3.
Submitter of study:_______________________________ (Signature)
Sponsor:_______________________________ (Signature)
Study Director:_______________________________ (Signature)
Page 3 of 7
-------
Certification of Availability of Raw Data
I hereby certify that the submitter possesses or has access to the
raw data used in or generated by the study summarized in this
document.
Submitter’s Representative:
signature/Date:
Typed Name:
Title:
Certification of ACCUraCY of Summary and AdeauaCV of the Study
I certify, in com 1ianCe with FIFRA section 4(e)(l)(A), that this
summary accurately represents the data presented in the report(s)
of this study cited by MRID, and that this study fully satisfies
all pertinent requirements of the OPP Guideline it addresses.
Submitter’s Representative:
S igriature/ Date : -
Typed Name:.
rrjt 1
Page 4 of 7
-------
171-4(a) Naturc Of The Residuc - Plants Study
ACCEF ANCE CRITERIA
Does vur study cet the following a piaacc criteria?
1. j ,. Pesticide radiolabeled in non.labile portion of molecule (tritium label strongly discouraged)
2. ‘I Specific activity sufficient to permit detection of low residue levels (0.01-0.1 ppm range)
3. .... L. Radiochemically pure grade of active ingredient
4. Pesticide applied to plant in manner simulating expected use
S. Total radioactivity measured in plant parts used for human food or animal feed
6. .j .. . .. Extractability of residue into solvents determined
7. Most of radioactivity extracted or exhaustive attempts (acid, base, enzyme) made to do so
& L... Major components or portion of the terminal residue identified (preferably by at least two
techniques-e.g., TLC, HPLc, MS)
Criteria marked with a ‘ are supplemental and may not be required for every study.
0
17 1.4(a)
December 24, 1989
Page 5 of 7
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171-4(a) Nature of the Residue: Plants Study
SUMMARY OF STUDY
1. Chemical structure and name of test material with site of
radiolabe]. indicated.
N-methyl-3-bromoanjljne; also known as “HERBICIDE”
The 14-C radiolabel was uniformly distributed through the
phenyl ring. (See cross-reference number 1 in Confidential
Attachment).
2. Specific activity of test material and detection limit
achieved for treated commodities.
Specific activity = 19.]. mci/mg. The detection limits for
treated soybean crop parts were 0.01—0.03 ppm.
3. Radiochemica]. purity of test material.
The radiochemical purity of ‘ 4 C-HERBICIDE was 98.5%.
4. Description of formulation and application of teSt material
and comparison of application (rate, timing in terms of growth
stage of crop) to expected use of pesticide.
Radiolabeled HERBICIDE was formulated as an emulsifiable
concentrate by addition of the emulsifier and organic solvent
used in the commercial product. The formulation was diluted
with water and applied to the soybean foliage at the fourth-
trifoliate stage at a rate equivalent to 0.25 pounds active
ingredient per acre, twice the rate permitted on HERBICIDE
labels. Soybeans would typically be in third to fourth-
trifoliate stages when HERBICIDE is applied.
5—7. Description of procedures used to quantitate and extract
radioactivity.
Total radioactivitjes of all plant parts were determined by
combustion of aliquots to ‘ 4 C0 2 . Samples were washed with
hexane followed by blending and extraction with 20% methanol
in acetone. In the case of dry crop parts, refluxing with lN
NC ]. was used to release additional residues. The total ppm
found in plant parts and the distribution of the residues in
the various solvents are summarized at the top of the next
page.
Extraction of the radioactivity in soybean seeds was not
attempted due to the very low level of residue (<0.01 ppm).
8. Description of procedures used to identify radiolabeled
residues.
Hexane washes and acetone extracts were analyzed directly by
Page 6 of 7
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Crop Dart
Green plant
Dry leaves
Dry stalks
Pods
PHI=preharvest
equivalents
2
75
75
75
tota).
hexa acetone
iN HC].
released
27.5
75
20
——
5
13.8
15
44
28
13
2.9
10
50
23
17
0.7
3
35
40
22
interval
in
days; 14-C
in
thin layer chromatography and autoradiography. Rf values were
compared to authentic standards. The HC1 solutions were
extracted with ether and the latter examined by TLC also. The
identities of spots were further confirmed by scraping the
spots from the plates and examining them by mass spectrometry.
The residues on each crop part are summarized in the following
table in which “TRR” is total radioactive residue, “4-hydroxy”
refers to N-methyl—3—bromo-4-hydroxyanjljne and “desniethyl”
refers to 3—bromoanjljne.
Crop part
Green plant
Total
14-C
%
of
Parent
72%
4-hvdroxy
12%
Desmethyl
9%
Unidentd
7%
27.5
ppm
Dry leaves
13.8
ppm
24%
39%
11%
26%
Dry stalks
2.9
ppm
18%
42%
8%
32%
Pods
0.7
ppm
15%
27%
15%
43%
In all cases about half the unidentified residue was activity
that could not be extracted by solvents or HC1. The remainder
consisted of very polar materials based on their TLC behavior.
Page 7 of 7
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CONFIDENTIAL ATTACHMENT
Phase 3 Summary of MRID 888888;
Metabolism of “HERBICIDE” in Soybeans: Project 23489
Guideline Reference:
171—4 (a)
Nature of the Residue:
Plant Study
Summary Prepared By:
A. Chemist
November 16 , 1980
Original Study Prepared By:
XYZ Laboratory
Chicago, IL
Page 1 of 3
-------
SAMPLE SUMMARY FOR RESIDUE CHEMISTRY STUDIES
Cross Reference Number 1
This cross reference number is used in the study in place of
the following words at the indicated page and line references.
DELETED WORDS: (identitiy of inert ingredients)
PAGE LINE REASON FOR THE DELETION FIFRA REFERENCE
6 4 Identity of inert ingredient 10(d) (1) (C)
Page 2 of 3
-------
SUPPLEMENTAL STATEMENT OF DATA CONFIDENTIALITY CLAIMS
[ AREA FOR SPECIFIC INFORMATION
SUPPORTING A CLAIM
OF CONFIDENTIALITY NOT
BASED ON
FIFRA 10(d) (1) (A), (B), OR (C))
Signature/D . —
Typed Name:
Title: —
Page 3 of 3
-------
December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
APPENDIX A SAMPLE SUMMARIES
-------
December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
-------
December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
-------
Phase 3 Summary of MRID 111111;
Chemical A - Fish Early Life Cycle - Rainbow Trout: Project 99333
Guideline Reference:
72—4 Fish Early Life Stage Test - Rainbow Trout
Bummary Prepared By:
A. Researcher
November 8, 1980
Original Study Prepared By:
Field Research, Inc.
Cincinnati, Ohio
Page 1 of 12
-------
Statement of No Data Confidentiality Claims
No claim of confidentiality is made for any information contained
in this study on the basis of its falling within the scope of FIFRA
section 10(d) (1) (A), (B), or (C).
Company:
Company Agent
Signature:
Title
(Typed Name)
(Typed Name)
Date:
Page 2 of 12
-------
Good Laboratory Practtce Statement
This study meets the requirement for 40 CFR Part 160
Submitter of study:_____________________________
Sponsor:_______________________________
Study Director: -
(Signature)
(Signature)
(Signature)
Page 3 of 12
-------
Certification of Availability of Raw Data
I hereby certify that the submitter possesses or has access to the
raw data used in or generated by the study summarized in this
document.
Submitter’ s Representative:
Signature/Date:___________________________________________
Typed Name:________________________________________
Title:______________________________
Certification of Accuracy o Summary and Adequacy o the Study
I certify, in compliance with FIFRA section 4(e)(l)(B), that this
summary accurately represents the data presented in the report(s)
of this study cited by MRID. Although this study does not fully
- satisfy the requirements of the OPP Guideline it addresses, it
should nonetheless be accepted in satisfaction of this Guideline
requirement. The specific deficiencies of the study and the
rationale for its acceptance in spite of those deficiencies are
individually identified and discussed within this summary.
Submitter’ s Representative:
Signature/Date:___________________________________________
Typed Maine:________________________________________
Title:
Page 4 of 12
-------
SAMPLE SUM 1ARY FOR ECOLOGICAL EFFECTS STUDIES
Subdivision E
Guideline Ref. No. 72-4
December 24, 1989
72-4 Early Lilc Stages Test
ACCEPTANCE CRITERIA
Does your study mcct thc following acccptancc critcria?
GENERAL INFORMATION
I. ...Y.... Date of est initiation and termination reported
MATERIALS/METHODS
2. Preparation of test solution (control/treatment) described
_I_ Any vehicle used to dissolve test material
..j.... If used, amount of vehicle added to control
_X_ Method used to get test material into solution was descnbed
_• Water source
•.. _ Water solubility of test material reported in ppm
_L_ Total amount of test material used
Temperature of water reported
Salinity (o/oo), if estuarine species is tested
3. Test fish
_ .. One of the following -organisms was tested: brook trout, rainbow trout. coho salmon,
chinook, bluegill, brown trout, lake trout, northern pike, fathead minnow, channel
catfish, sheepshead minnow, silverside
°.1.... History of test organisms described (strain, diseases and treatment)
J Health described (sickness; injuries; abnormalities; name of medication, if used;
pretest diet, number of days brood fish were quarantined)
4. Size/age/physical condition
..X... Age of unfertilized eggs and milt at the beginning of test
Procedures used to strip eggs and milL described
j Procedures used in storage and handling of eggs described
....L... Fertilization process described
Sourvei cclimazion described -
Complete name and address of test fish supplier, source of food and dilution water, size of
test container in milliliters (mL), temperature, p 1-I, dissolved oxygen, name of equipment used
to measure water quality, feeding schedule, holding, acclimation period, test organisms are
from same source, number of days held, salinity (0/00)
Criteria marked with a • are supplemental and may not be rcquired for every study.
Page 5 of 12
-------
Subdivision E
Guideline Ref. No. 72.4
December 24, 1989
6. Composition and size of test vessels described
Y Material type
V Volume in liters (L) or size in centimeters (cm)
_ Depth of test solution in cm or volume of dilution water (L)
7. Test system descnbed
( Source of dilution water
j_. If flow-through, system and flow rate/day described
_j_ Procedures used to prepare toxicant stock solution
• j_ Criteria used to determine effects
& Test design followed
.1.... Method used in assigning test organisms to test and control groups and number of
replicates used
1’ Number of eggs per dose level and control group (80)
•j Name of protocol followed dunng this test
_j_ Loading (weight per unit volume of water)
j 5 treatments and 1 control used
j Length of exposure period
_. Type of control (positive, negative, or solvent control)
•j_ Temperature
LpH
°4 •_ Lighting (tune in hours and intensity - footcandles)
Dissolved oxygen
_J _ Water hardness
J Alkalinity
•J _ Conductivity (umhos/cm)
j . Range finding test results (concentrations and mortality) if conducted
j... Water: physical characteristic at the end of test . water depth and volume
_ _ Period food was withheld prior to termination
:....... Method used to keep eggs clean described
DISCUSSION AND RESULTS
9. i L MATC value reported in ppm. Raw data provided.
10. .1.... Any precipitation and solubility problems described.
11. j ._ Statistical method referenced.
12. .j.. Discussion of concentration(s) producing intoxication provided.
Water physical characteristic at the end of test . temperature, pH, and dissolved
oxygen
Concentration analysis (if flow through)
Criteria marked with a are supplemental and may not be required for every study.
Page 6 of 12
-------
J Subdivision E
Guideline Ret No. 72-4
December 24, 1989
13. .. . . . . . Signs of intoxication noted.
Number of embryos hatched per replicate reported
_ . _ Time to hatch per replicate reported
Mortality of embryos, larvae, and juveniles per replicate reported
_ Time to swim-up per replicate reported
..1_ Measurements of growth-weight and length per replicate reported
_ . . _ Fish physiology, locomotion, behavior and pathology reported
_ _ When larvae fish released from the retaining cup, % complete or hours elapsed
identified
14.// No-observed-effect level, if determined.
15. The relationship between the results of the study and the test chemical described, including
. ., .. Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
Rationale for minor deviations from the protocol
16.’j . . Graphs, printouts and other calculations are attached to report
Criteria marked with a * are supplemental and may not be required for every study.
Page 7 of 12
-------
72-4 Fish Early Life Stage Test
Identification of Test Substance
Chemical A Technical; Batch No. 13-10-10-32; 93% active
ingredient; a tan powder.
General Information
1. Conclusions:
The MATC of Chemical A Technical for rainbow trout
( Salmo gairdnerj ) was < 0.062 mg AI/L based on mean
measured concentrations.
Test initiation: 2/14/87 Test termination: 5/13/87
Materials/Methods
2. Description of test solution (control/treatment)
preparation, including:
a. Maximum solubility of 0.343 mg Chemical A/L was
achieved after mixing for 48h in 0.05% acetone.
Solubility of Chemical A in water (20° C) is 0.35
ppm.
3. Description of test species
a. Rainbow trout ( Salmo gairdnerj )
4. Description of source/acclimation, including:
a. Test fish were obtained from the Mt. Lassen trout
farm, Red Bluffs, California.
b. Sperm and eggs were received and fertilized at 8
C and gradually over 1 hour acclimated to test
temperature (12° C).
5. Description and size of test vessels
Each glass aquarium measured 25 x 16 am with a water
depth of 24 cm, yielding an approximate 9.6-liter
replicate-chamber volume. The rainbow trout eggs were
incubated in cups suspended in the treatment and control
water. These egg incubation cups were made from 7-cm
diameter tubing with stainless steel screening (16 mesh)
fused to the bottom.
Page 8 of 12
-------
b. The diluter delivered an average rate of
approximately 63 mL/minute/replicate of test
solution or control water to the test vessels which
was sufficient to replace a replicate volume 9.4
times in a 24-hour period.
c. The following parameters were studied: embryo
viability, survival of organisms at hatch, lay 1
survival, and growth. Growth, as determined ;
standard length of the fry, was determined by
photographic method of McKim and Benoit (1975)
study day 49. At test termination, study day 74
days post-hatch), all surviving fish were measur i
for standard length and wet weight.
7. Description of test design
a. A control, solvent control, and five nominal
Chemical A Technical concentrations of 0.06, 0.12,
0.25, 0.50, and 1.0 mg/L based on active ingredient
were tested. The solvent control solution contained
the maximum amount of DMF present in any test
concentration (0.014 mL/L). Thirty rainbow trout
eggs were randomly introduced into each replicate
chamber (60 eggs per concentration).
b. The test chambers were immersed in a temperature
controlled water bath held at 12 ± 2°C. During the
testing period, the rainbow trout fry were on a 16-
hour daylight photoperiod with the eggs shielded
from excess U.V. light exposure. The light
intensity was 158 ± 25 foot—candles at the water
surface.
c. The well water characterized as having a H of 7.8 -
8.3.
d. When hatching commenced, the number of eggs hatched
in each incubation cup was recorded daily until
hatching was completed. The 60-day post-hatch
growth period began when hatching was greater than
95 percent (Day 14). On study-day 27 (13 days post-
hatch), the rainbow trout sac fry were removed from
the duplicate egg incubation cups and separated into
4 replicate growth chanbers per concentration.
Survival and fry growth data were collected on Days
35 and 60 post-hatch. Feeding began on Day 27 (13
6. Description of test system
a. Dilution water was aerated well water.
Page 9 of 12
-------
days post-hatch). The fry were fed brine shrimp
nauplii three times each day. Commercial fish food
was added to the diet as the fish became larger.
8. Discussion and Results
a. One-way analysis of variance (ANOVA) calculations
were used to determine if significant differences
existed between the control and treatment levels.
If treatment effects were indicated by a significant
F—test of the mean square ratios, Tukev’s HSD
multiple means comparison test was used to determine
which exposure levels differed from the control
values.
b. The mean measured concentrations of Chemical A
Technical were 0.062, 0.097, 0.20, 0.41, and 0.84
trig AI/L. The mean measured concentrations ranged
from 80 to 103% of the nominal test concentrations.
c. Based on the length and weight data from this 60-
day post-hatch rainbow trout ( Salmo gairdneri ) early
life state study, the Maximum Acceptable Toxicant
Concentration (MATC) of Chemical A Technical was
estimated to be < 0.062 mg AI/L, the lowest
concentration tested.
d. Morphological and behavior abnormalities such as
curved spine, fish resting on the bottom of the test
chamber, loss of equilibrium, light discoloration,
quiescence, enlarged yolk sac, and fishing swimming
vertically in the test chamber were scattered
throughout the controls and the three lowest test
levels in a small percentage of the fish. By Day
50 (36 days post—hatch), most of these effects had
either disappeared or the affected fish had died.
Quiescence and fish resting on the bottom of the
test chamber persisted in a few fish in the 0.20 trig
AI/L concentration until just before test
termination.
Results for the effect of Chemical A Technical on
length of rainbow trout are shown in Table 1
(attached). The single surviving fish in the 0.41
mg AI/L concentration was not included in the 35-
day post-hatch length analysis since a group that
contains only one fish cannot be compared with other
groups each containing several fish in an analysis
of variance. The 0.20 mg AI/L concentration fish
Page 10 of 12
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SAMPLE SUMMARY FOR ECOLOGICAL EFFECTS STUDIES
were the only test fish that showed a significant
reduction in length when compared to the control at
35-days post-hatch. By 60-days post-hatch, the fish
in the lowest three test concentrations all showed
a significant (P< 0.05) length reduction when
compared to the control.
Results for the effect of Chemical A Technical on
wet weight of rainbow trout are shown in Table 1
(attached). Growth of the trout fry, as measured
by wet weight after 60 days of exposure to Chemical
A Technical, was significantly reduced in the lowest
three test concentrations.
9. Discussion relating results of study to test chemical
a. Based on growth (weight and length), the Maximum
Acceptable Toxicant Concentration (MATC) of Chemical
A Technical was estimated to be < 0.062 mg AI/L, the
lowest concentration tested.
b. Justification of deviation from acceptance criteria:
although neither a NOEL nor a defined MATC was
determined in this study, when these results are
included with those of our previously submitted Fish
Early Life Stage study on Chemical A, the Guideline
requirement has been satisfied.
Page 11 of 12
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Table 1.
Summary of statistical analysis of embryo viability,
survival of embryos at hatch and survival, total length,
and wet weight of rainbow trout ( Sa]jno gairdneri ) exposed
to Chemical A for 88 days (60 days post-hatch).
Mean
Measured
Conc.
(mg/ L)
Embryo
Viability
(%)
Survival of
Organisms at
Hatch (%)
Larvae (60 days Post-hatch )
Larvae Mean Total Mean Wet
Survival Length Weight
(%) (mm) (g)
.840 Mean
.410 Mean
.200 Mean
.097 Mean
.062 Mean
Solvent
Control Mean
Control Mean
Pooled Controls
(In this sample, values have been omitted)
Page 12 of 12
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Phase 3 Summary of MRID 22222 and Related MRID 232323;
Chronic Feeding of “HERBICIDE” in the Rodent: Project 23456
Guideline Reference:
83-1 Chronic Feeding in the Rodent and Nonrodent
Summary Prepared By:
J. Morrison, PhD.
March 15, 1980
Original Study Prepared By:
tJnicorp Group Laboratories, Inc.
Providence, RI
Page 1 of 8
-------
Statement of
No Data Confidentja].jtv Claims
No claim of confidentiality is made for any information contained
in this study on the basis of its falling within the scope of FIFRA
section lO(d)(l)(A), (B), or (C).
Company:
Company Agent:
Title:
(Typed Name)
(Typed Name)
Signature:
Date :
Page 2 of 8
-------
Good Laboratory Practice Statement
This study meets the requirement for 40 CFR Part 160
Submitter of study:______________________________
Sponsor:_______________________________
Study Director:______________________________
(Signature)
(Signature)
(Signature)
Page 3 of 8
-------
Certification of Availability of Raw Data
I hereby certify that the submitter possesses or has access to the
raw data used in or generated by the study summarized in this
document.
Submittert s Representative:
Signature/Date:______________________________________________
Typed Name:
Title: ______________________________
Certification of Accuracy of Summary and Adequacy of the Study
I certify, in compliance with FIFRA section 4(e)(l)(A), that this
summary accurately represents the data presented in the report(s)
of this study cited by MRID, and that this study fully satisfies
all pertinent requirements of the OPP Guideline it addresses.
5 Representative:
Signature/Date:___________________________________________
Typed Name:
Title:
Page 4 of 8
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Subdivision F
Guideline Rer. t 4o. 83-i
December 24, 1989
ACCEPTANCE CRITERIA
ur study mxt thc following a piana aiicria?
1. • _ Technical form of the active ingredient tcstcd.
2. j_ At teast 20 rodents or 4 nonrodents/sex/group ( 3 test groups and control group).
3. •j•••_ Dosing duration in rodents minimum 12 month nonfood use, 24 months food use; in
nonrodents minimum 12 months’.
4. _j_ Doses tcstcd includc signs of toxicity at high dusc but no lethality in nonrodents or a limit
dose if nontoxic (1,000 mg/kg).
Doses tested include a NOEL
6.’_ Analysis for Lest matenal stability, homogeneity and concentration in dosing medium
7. _ . _ Individual daily observations.
8. .. .j. Individual body weights.
9. ....... . Individual or cage food consumption.
10.4...... Opihalmoscopic examination (at least pertest and at term) control and high dose.
11. 4_ Clinical patholo data for all nonrodenis and at least 10 rodents/group consisting of 12, 13
&14.
12. 4..... Hematolo ’ at 6 month intervals consisting of at least;
.4.. Ezythrocyte count j. Leucocyte count
_. . _ Hemoglobin ‘j , Differential count
_..X_ Hematocnt _.!L Platelet count (or clotting measure)
13. Clinical chemistry at 6 month intervals consisting of at least;
..... L Alkaline phosphatase — Total Protein
— Aspartate aminotransferase Albumin
_• Alanine aminotransferase .4.... Urea nitrogen
•_ Creatinine kinase _• _ Inorganic phosphate
‘ Lactic dehydrogenase Calcium
.j.. Glucose ‘J_ Potassium
j_ Bilirubin _L Sodium
L Cholesterol Chloride
* ‘ Creatinine
14. J . .... Urinalysis at 6 month mtervals consisting of at least;
.. . j... . Blood Total bilirubin
_..L. Protein ‘ . . . L Urobilirubin
.j_ Ketone bodies — Sediment
.. . L, . . Appearance ‘1 Specific gravity (osmolality)
4... Glucose Volume
is. . . L .. Individual necropsy of all animals.
Criteria marked with a • arc supplemental and may not be required for every study.
&-1 Chronic F ling in the Rodent and Nonrodcnt
Page 5 of 8
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SAMPLE SUMMARY FOR TOXICOLOGY STUDIES
• . Subdivision F
Guideline Ref No. 83-1
December 24, 1919
16. L l-listopathology of ihe following tissues performed on all nonrodents and rodents, all control
and high dose animals, all animals that died or were killed on study, aU gross lesions on all
animals, target organs on aft animals and lungs, liver and kidneys on all other animals.
— aorta _ _ jejunurn peripheral nerve
.j eyes bone marrow kidncysf
_• _ caecum livcrf 4 esophagus
• j. . colon _y_ lung jL ovaries
duodenum _ , lymph nodes oviduct
brain . . .L stomach _ .. pancrcas
— skin 4... . mammary gland — rectum
L. heart •...L spleen _ _ spin.il cord ox)
. .. . . . . . testes _ _ musculature thyroid / parathyroids
.... . L.... pituitary 4. . . ... epididymis L. salivary glands
_L. ileum _ adrenals _• _ thymus
_ _ trachea L urinary bladder .4 . .. accessory sex organs; uterus
gall bladder
t organs to be weighed
In some cases, a six month study may be acceptable. Contact EPA to discuss the criteria for
determining if such a study cart be used to support reregistration.
Criteria marked with a * are supplemental and may not be required for every study.
Page 6 of 8
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83-1 Chronic Feeding Study in the Rodent and Nonrodent
1. Technical PESTICIDE (96.2%) was used.
2. Weaning SPF Sprague-Dawley rats (75/sex/dose) were fed diets
containing 0, 100, 300 or 1,000 ppm technical pesticide.
3. Duration was 24 months.
4. At 1,000 ppm, the following effects were seen: increased
mortality in females, (months 18.24); decreased weight gain
in males (—15%) and females (—22%); ataxia, tremors,
urogenital staining and lacramation in males and females;
other effects in clinical chemistry and histopathy and will
be discussed in those sections.
5. The 100 ppm dose levels was a NOEL for all parameters
measured. -
6. Test diet, prepared biweekly, was stable for 3 weeks with
concentrations of 98.5±2, 101.3±3 and 96.2±2% for the test
doses of 100, 300 and 1,000 ppm.
7. Increased mortality was observed in the high dose females in
months 18-24. Signs of toxicity, including ataxia, tremors,
urogenital staining and excessive lacrimation were seen in the
1,000 ppm males and the 300 and 1,000 ppm females. NOELs for
all observations were 300 ppm for males and 100 ppm for
females.
8. A dose—related decrease in weight gain was seen in the 300 and
1,000 ppm females and the 1,000 ppm males.
9. Dose Food Consumption PESTICIDE dose
Ippm) ( g/Kg) ( mg/Kg/day )
__ F M F
0 53.0 68.9 0.00 0.00
100 52.1 68.1 5.00 7.00
300 52.2 65.8 15.75 19.80
1000 54.8 77.2 54.69 77.19
10. Opthalmoscopic exams were not carried out, however since no
adverse pathological lesions were noted in this study and the
long—term rat and mouse study, it is doubtful that these
opthalmoscopjc exams would add much to the experiment.
11. Complete clinical pathology measurements were carried out on
10 rats/sex/dose at 6, 12, 18 and 24 months.
12. There were no dose-related effects on any hematogical
measurements.
13. Clinical chemistry, as noted in the check list, showed
increased alkaline phosphatase and lactic dehydrogenase in the
females at 18 and 24 months at 1,000 ppm. No other effects
were noted. Total protein, aspartate aminotransferase and
Page 7 of 8
-------
SAMPLE SUMMARY FOR TOXICOLOGY STUDIES
calcium were not determined.
14. No dose-related effects were noted on any urinalysis
measurements. Although “sediment,” per Se, was not noted.
15. Gross necropsy of all rats showed stained/mottled liver in
females at 300 and 1,000 ppm and in males at 1,000 ppm.
16. Increased absolute organ weights of brain, liver and kidneys
were observed in females at 1,000 ppm whereas relative
organ/body weight ratios were increased for liver and kidney.
The only histopathological effect noted was necrosis in the
livers of females at 300 and 1,000 ppm and males at 1,000 ppm.
Histopathological exams of the aorta, skin and rectum were not
carried out.
17. Deficiencies and their impact:
It is our opinion that the lack of measurements of the
3 parameters listed in criteria 13 have little affect on the
final determination of the study. Other serum ertzytnes showed
no adverse effects and histopathological exams were adequate
to detect any abnormalities that would be signaled by changes
in these parameters
It is our opinion that, although no specific observation
for “sediment” was reported, that the comments and
observations under appearance would be sufficient to detect
the appearance of sediment in any urine samples.
Although histopathology exams of the aorta, skin and
rectum were not done, gross observations failed to detect any
significant abnormalities.
Page 8 of 8
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Phase 3 Summary of MRID 789878 and Related MRIDs 123212 and 234567;
Makebelonate - Hydrolysis Study: Project 890234
Guideline Reference:
161-1 Hydrolysis
Summary Prepared By:
Henly Hankboro
November 15, 1980
QRIGINAL STUDY PERFORMED BY
Curie Chemistry Associates
Shreveport, LP
Page 1 of 9
-------
Statement of No Data Confidentiality Claims
No claim of confidentiality is made for any information contained
in this study on the basis of its falling within the scope of FIFRA
section 10(d) (1) (A), (B), or (C).
Company
Company Agent
Title
(Typed Name)
(Typed Name)
Signature:
Date:
Page 2 of 9
-------
Good Laboratory Practice Statement
This study meets the requirement for 40 CFR Part 160
Submitter of study:______________________________
Sponsor:_______________________________
Study Director:_______________________________
(Signature)
(Signature)
(Signature)
Page 3 of 9
-------
Certification of Availability of Raw Data
I hereby certify that the submitter possesses or has access to the
raw data used in or generated by the study summarized in this
document.
Submitter’ s Representative;
Signature/Date:___________________________________________
Typed Name:_______________________________________
Title: ______________________________
Certification of Accuracy of Summary and Adequacy of the Study
I certify, in compliance with FIFRA section 4(e)(l)(A), that this
summary accurately represents the data presented in the report(s)
of this study cited by MRID, and that this study fully satisfies
all pertinent requirements of the OPP Guideline it addresses.
Submitter ‘ s Representative:
Signature/Date:___________________________________________
Typed Name:________________________________________
Title:______________________________
Page 4 of 9
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SAMPLE SUMMARY FOR AN ENVIRONMENTAL FATE STUDY
Subdivision N
Guideline Ref No. 16 1-1
Occember 24, 1989
161-1 Hydrolysis
ACCEPTANCE CRITERIA
Docs your study mccl the following a p1an aitcria?
1. J . Test substance was the Technical Grade (TGA [ ) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2. ... L Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeled).
3. L. The positions of the Al radiolabeling were appropriate (if radiolabeted).
4. j _ . Solubility of the Al at each pH tested.
5. .... L. Concentration of AL (in solution) within the known solubility range of the Al and below
250 ppm
6. Study was conducted in darkness.
7. .... L. Temperature held at s±i°c.
8. j ... . Sterility of glassware, reagents, chemicals, etc. was assured.
9. V latiles (if any) were trapped.
io. . . L . . . . Solutions were buffered.
11. .. . . Cosolvent (if any) did not exceed 1%.
12. j... . pH of each test solution was reported and was correct (nominally 5,7 and 9) and constant.
13. .j. ... Sampling intervals were adequate.
14.’..... L. Study was conducted for one half-life or 30 days.
15, A reasonable attempt was made to identify alt degradatesi1 ydrolysaIes which exceeded 10%
(yield) of the initial concentration of the Al.
16. 4— Material balance was reasonable (>90% .
-------
19. j. .. }-iaIf-live3 were lculatcd.
Subdivision N
Guideline Ref. No. 161.1
December 24. L989
Criteria marked with a ‘ are supplemental and may not be required for every study.
Page 6 of 9
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SUMMARY OF CONCLUSIONS
Makabelonate hydrolysis Tl/2:
PH5 PH7
120 da 88 da
TEST MATERIAL
Conclusion:
IUPAC designation: 1, l -tetracarbamo—3-trjchloro_nucleosjde
CAS Number:
Source:
Lot Number:
Formulation and Purity:
Chemical class:
Solubility:
Radioactive labeling site:
Specific activity:
999g9—99—0
Synthetic Chemical Company
l23456—DDT
TGAI, 94% Al, no identifiable
impurities
01
100 mg/mL
Uniform, furocpuro ring
600 uCi/minole
Lighting conditions:
Temperature range:
Spiked samples:
Water and characteristics:
Buffers used:
Coso].vent:
Sampling intervals:
Volatiles trap:
Sample extraction;
Extraction efficiency:
Complete darkness
20 +
10, 100, 1000 mg/L efficiency = 98%
deionized water, filtered through a
0.22 micron Millipore filter, char-
acteristics: pH=7.8; hardness, 140 ing
as CaCO 3 ; alkalinity, 14 mg as CaCO 3 ;
and conductivity, 700 uxnho.
pH 5, 0.1 n J acetate buffer
pH 7, 0.1 n)j phosphate buffer
pH 9, 0.1 m borate buffer
None
Time zero, 1, 3, 7, and 14 days.
Thereafter, samples were taken every
2 weeks.
A trap consisting of hyaminehydroxjde-
saturated filter paper was used to trap
radioactive volatiles.
Parent and degradate were extracted
using a guideline-approved
acidification method and dried on
filter paper.
Parent, 97%; Degradate, 97.5%
HIDROL 1 YSIs SUMMARY SHEET FOR MAKABELONATE (STOMPOUT)
IDELINE REFERENCE NO: 161-1
Stable
pH9
72 da
TEST METHODOLOGY
Page 7 of 9
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Counting method:
RESULTS
Materials balance
Samples on filter paper were stored at
-20 °C for up to a week before
counting. Stability studies have shown
that the samples are stable in this
condition for up to a month.
Filter paper was immersed in PPO-POPOP
cocktail and counted in a Packard
Tricarb scintillation counter.
- experimental
Parent - 98.4% of the added material
Degradate - 98.4% of the added material
- Spiked samples
Parent — 99.7% of the added material
Degradate - 99.7% of the added material
Tl/2 Determination method: Linear Regression — SYSTAT model
REGISTRANT CAN
PUT
GRAPHS
OR
TABLES
HERE
AT THEIR OPTION
Sample stability:
Materials balance
Page 8 of 9
-------
DISCUSSION
AREA FOR
OPTIONAL
EXPLANATION
OF
STUDY DEVIATIONS
End
Page 9 of 9
-------
December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
APPENDIX B: PR NOTICE 89-3
-------
December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
-------
PR NOTICE 89-3
NOTICE TO REGISTRANTS SUBJECT TO
REREGISTRATION PHASE 3
Attention: Persons responsible for preparing Reregistration
Phase 3 responses.
Subject: Format standards for Reregistration Phase 3 sumrna-
ries and reformatted versions of studies previously
submitted under the Federal Insecticide, Fungicide
and Rodenticide Act (FIFRA) and certain provisions
of the Federal Food, Drug, and Cosmetic Act (FFDCA)
I. Purpose
To define standard formats for summaries and reformatted ver-
sions of previously submitted studies, as required to be submitted
in Phase 3 of reregistration by section 4(e) (1) of FIFRA, as
amended.
II. Applicability
This notice applies to all summaries of previously submitted
data provided to the Agency under section 4(e)(1)(A) or (B) of
FIFRA, and to all reformatted versions of previously submitted data
provided under section 4(e)(1)(C) or (D) of FIFRA.
III. Effective Date
This notice is effective upon issuance.
IV. Background
The amendments to FIFRA of October, 1988 define a new five-
phase program of reregistration. In the third of these five phases
registrants of pesticides first registered before 1984 must submit
structured summaries of all previously submitted studies on which
they wish to rely to support reregistration, and must submit
-------
complete reformatted versions of selected tOxicology and residue
studies as well.
The schedule for reregjstra j 0 laid out in the amended Act
requires that many thousands of these documents will arrive at the
Agency over a very short time, and requires further that the Agency
review them very soon after they arrive.
Thus it is critical to the Success of the overall reregistra...
tion program that these summaries and reformatted Studies be added
to the Agency’s master document management Systems as rapidly and
economically as possible. Toward that end these standards have
been defined.
V. Relationship of this Notice to Other Policy and Guidan
This notice complements PR Notice 86—5, but does not super-
sede it. PR Notice 86-5 remains controlling with respect to all
data submissions to OPp exce those associated with Reregjstra 10
Phase 3.
This notice also complements the guidance called for in sec-
tion 4(e)(4)(A) of amended FIFRk, which specifies content-related
requiremen 5 for reregistra j 0 Phase 3 responses.
VI. Scope and Organization of the Phase 3 Submittal Pack g
The scope of a single Phase 3 submittal must be limited to a
single regjstran I 5 submission for a single chemical within a
single reregistra i 0 case. A Phase 3 submittal package Consists
of the following four elements, Physically arranged in the order
listed:
A. One Signed and dated copy of the “Reregistra j 0 Phase 2
Chemical Response Worksheet”, which serves as a transmittal
document for all other components of the response (be sure not
to include any information you claim as Confidential in this
document),
B. One copy of each summary of a Previously submitted study shown
to be ‘provided’ on the Worksheet/Transmittal, formatted as
described in section VIII below,
C. One copy of each reformatted version of a Previously submitted
study shown to be ‘provided’ on the Worksheet/Transmittal,
formatted as described in section IX below, and
D. One copy of any other correspondence or material submitted
with the above elements in the package. (See the Phase 3
Guidance for more information about specific documents which
2
-------
would fall into this “other material” category, such as waiver
requests and supporting rationales, discussions of applicabil-
ity of individual data requirements, and the like.) Generally
these documents should not need to contain any confidential
business information. If you wish to make a claim of confi-
dentiality for any information in any of these documents, the
specific information covered by the claim and the basis for
the claim must both be clearly identified.
In other words, a Phase 3 submittal consists of the worksheet/
transmittal, followed by ill the summaries, followed by jJ, the
reformatted studies, followed by any other materials. This
arrangement is illustrated in Attachment 3.
VII. Scope of a Study
The scope of one Phase 3 summary, or of one reformatted
report, is a single study. As defined in PR Notice 86-5, a “s tudy”
is the report of a single scientific investigation, including all
supporting analyses required for logical completeness. In
determining the scope of a study for Phase 3 responses, take
particular care to include all significant related documents,
including supplements, addenda, supporting analyses and the like,
even if they were previously submitted at a different time from
the primary report of the study. One of the objectives of the
requirement to summarize and reformat studies is to bring together
all such partial or fragmentary reports of studies into well
defined, logically complete new documents. Take care as well to
summarize and reformat separately logically distinct studies which
may have originally been submitted bound together in a single
volume.
In most cases a study will correspond in scope to a single
Guideline requirement. The following special considerations,
however, deserve mention; note that these points reflect the same
principles of study identification originally laid out in section
C(l) of PR Notice 86-5.
When a single study (i.e., the report of a single investiga-
tion) addresses more than one Guideline topic, a separate summary
should be prepared for each Guideline topic addressed. Each such
summary derived from a single original study would, of course, cite
the same original MRID.
In the case of product chemistry data, the scope of a single
‘study’ may be broadened when appropriate to include all guideline
requirements within a single guideline series (such as series 61,
62, or 63.)
In the case of residue data, the scope of a single ‘study’
should be limited to a single analytical method, an investigation
3
-------
of the nature of the residue in a single plant or animal species,
or to an investigation of the magnitude of residues resulting from
treatment of a single crop or from processing a single crop. This
is true for residue data originally submitted in broad, multiple
topic volumes as well as for residue data originally submitted in
more narrowly defined studies.
When multiple field trials are associated with a single inves-
tigation, the reports of all such trials should be treated as a
single study.
VIII. Required Elements for Phase 3 Summaries
Most of the elements required for Phase 3 summaries are
defined and discussed in PR Notice 86—5. The specific requirements
for summaries and the order in which they must appear are as
follows. These elements and their order are illustrated in Attach-
ment 4.
A. Title Page. The first page of each summary must be a title
page. The title page must !2 include any information claimed as
confidential, and must include the following elements:
1. Title. The title of each Phase 3 summary must be in
standard form, to make these summaries easy to recognize from
citations. The required elements in the title are:
o the introductory words “Phase 3 Summary of MRID”,
o the MRID assigned to the previously submitted primary
report,
o i more than one previously submitted document is
included in the scope of the summary, the MRIDs assigned
to all other included reports,
o the descriptive title of the original study, and
o the performing laboratory’s or submitter’s study identi-
fication number or code, if any.
Represented schematically, with mandatory text in bold face,
unconditionally required elements in square brackets, and
conditional elements in curved brackets, the title of a Phase
3 summary would be:
Phase 3 Summary of MRID [ primary MRID] (and related t4RIDs
a, b, and C): (Descriptive Title]: (Project Nr 123).
4
-------
Here is an example of a title conforming to these require-
ments:
Phase 3 Summary of MRID 123456 and related MRIDs 141865
and 141866: Teratology of Technical Buggywhip to New
Zealand Rabbits: Project Number 82—104.
2. Guideline Reference Number and Name of Requirement.
Enter these elements on the title page exactly as they appear
on the Phase 3 Worksheet.
3. Author. Identify the author of the summary , not the
author of the original study being summarized.
4. Date. Include the date of the summary , not the date of
the original study being summarized.
5. Performing Laboratory. Enter the name(s) and address(es)
of all laboratories who performed or contributed to the origi-
nal study. If more laboratories were involved than can be
listed on the title page, include on the title page a refer-
ence to an internal page on which they all appear.
6. Pagination. Enter the phrase “Page 1 of x”, where “x”
is the total number of pages in the body of the summary,
exclusive of the confidential attachment or supplemental
statement of confidentiality claims, if any.
B. Statement of Data Confidentiality Claims. Page 2 of each
Phase 3 summary must include one of the two standard statements of
data confidentiality claims defined in section D(2) of PR Notice
86-5, either asserting that no claims of confidentiality under
FIFRA section l0(d)(l)(A), (B), or (C) are made for the summary,
or that information claimed confidential under that authority has
all been removed from the body of the summary to a confidential
attachment. Note that these statements apply only to confidential-
ity claims under FIFRA section lO(d)(l)(A), (B), or (C): that you
will have no further opportunity to register claims of confiden-
tiality not made at this time, and that the wording of the two
forms of the statement is mandatory . You may add to either state-
ment as appropriate, but the language specified must all be
included and must not be altered or contradicted. This statement
must be signed and dated by an authorized representative of the
submitter.
C. Statement of Good Laboratory Practice Compliance. Page 3 of
each Phase 3 summary must bear an appropriate statement concerning
the compliance or non-compliance of the original study with EPA’S
Good Laboratory Practices regulations (40 CFR 160).
Note that while a statement of GLP compliance is always re-
quired, its wording is not prescribed. Since many of the studies
5
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summarized in Phase 3 will have been performed before these regu-
lations were in place, a statement to that effect will often be all
that is necessary or appropriate. See 40 CFR 160.12 for more
information about this requirement.
tinder the regulations the GLP compliance statement must be
signed by the study director, the study sponsor, and the study
submitter. Clear, legible photocopies of statements from the
original study report are acceptable. If the study director or
study sponsor is unavailable to sign a new statement, this should
be explained in the statement, and it must be signed by the
submitter.
D. Certification of Access to Raw Data. Two mandatory statements
are required on page 4 of each Phase 3 summary. The first of them
is a certification of access to raw data. (The second is discussed
in paragraph E below.) The wording of the certification of access
to raw data is mandatory , and it must be signed and dated by an
authorized representative of the submitter:
Certification of Access to Raw Data
I hereby certify that the submitter possesses or has access
to the raw data used in or generated by the study summarized
in this document.
Submitter’ s Representative: [ Signature/Date]
[ Typed Name]
[ Title]
E. Certification of Accuracy of Summary and Adequacy of the
Study. The second certification required on page 4 of each Phase
3 summary is the appropriate one of the two following statements
with respect to the accuracy of the summary and adequacy of the
underlying study to satisfy Guideline requirements. Note that only
one of these two statements is appropriate, that the wording o ’
the certification is mandatory , and that either version must be
signed and dated by an authorized representative of the submitter.
1. For use when the study fully satisfies the requirement:
I certify, in compliance with FIFRA section 4(e)(l)(A), that
this summary accurately represents the data presented in the
report(s) of this study cited by MRID, and that this study
fully satisfies all pertinent requirements of the OPP Guide-
line it addresses.
Submitter’s Representative: [ Signature/Date]
[ Typed Name)
[ Title]
6
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2. For use when the study does not satisfy the requirement:
I certify, in compliance with FIFRA section 4(e) (1) (B), that
this summary accurately represents the data presented in the
report(s) of this study cited by MRID. Although this study
does not fully satisfy the requirements of the OPP Guideline
it addresses, it should nonetheless be accepted in satisfac-
tion of this Guideline requirement. The specific deficiencies
of the study and the rationale for its acceptance in spite of
those deficiencies are individually identified and discussed
within this summary.
Submitter’ s Representative: [ Signature/Date)
[ Typed Name)
[ Title]
P. Body of the Summary. This should be prepared as specified in
the Phase 3 Guidance material appropriate to the Guideline topic
addressed.
G. Confidential Attachment. The confidential attachment, if any,
and its cover sheet, as defined in section D(3) of PR Notice 86-5,
follow the body of the summary, within the same binding.
H. Supplemental Statement of Data Confidentiality Claims. A
supplemental statement of data confidentiality claims based on
authority other than FIFRA section lO(d)(l)(A), (B), or (C), if
any, as defined in section D(4) of PR Notice 86—5, follows the
confidential attachment to the summary, if any, or the body of the
summary if there is no confidential attachment.
IX. Required Elements for Studies Reformatted in Phase 3
The elements required for studies reformatted in Phase 3 are
very similar to those required for Phase 3 summaries. The specific
requirements and the order in which they must appear in each refor-
matted study are as follows. These elements and their order are
illustrated in Attachment 4.
A. Title Page. The first page of ea ch study must be a title
page. Do not include any information claimed as confidential on
the title page. Do include the following elements:
1. Title. The title of each study reformatted in Phase 3
must be in standard form, to make these studies easy to recog-
nize from citations. The required elements in the title are:
o the introductory words “Phase 3 Reformat of MRID”,
o the MRID assigned to the previously submitted primary
report,
7
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o if more than one previously submitted document is
included in the scope of the study, the MRIDs assigned
to all other included reports,
o the descriptive title of the study, and
o the performing laboratory’s or submitter’s study
identification number or code, if any.
Represented schematically, with mandatory text in bold face,
unconditionally required elements in square brackets, and
conditional elements in curved brackets, the title of a study
reformatted in Phase 3 would be:
Phase 3 Reformat of MRID [ primary MRID and related
MRIDs a, b, and C): [ Descriptive Title (Project Nr
123).
Here is an example of a title conforming to these require-
ments:
Phase 3 Reformat of MRID 123456 and related MRIDs 141865
and 141866: Teratology of Technical Buggywhip to New
Zealand Rabbits: Project Number 82—104.
2. Guideline Reference Number and Name of Requirement.
Enter these elements on the title page exactly as they appear
on the Phase 3 Worksheet.
3. Author. Identify the author(s) of the original study.
4. Date. Report the date of the original study.
5. Information about Reformatting. Report the date of the
reformat, and by whom it was done.
6. Performing Laboratory. Enter the name(s) and address(es)
of all laboratories who performed or contributed to the origi-
nal study. If more laboratories were involved than can be
listed on the title page, include on the title page a refer-
ence to an internar page number on which they all appear.
7. Pagination. Enter the phrase “Page 1 of X”, where “x”
is the total number of pages in the body of the study,
exclusive of the confidential attachment or supplemental
statement of confidentiality claims, if any.
B. Statement of Data Confidentiality Claims. Page 2 of each
Phase 3 study must include one of the two standardized statements
of data confidentiality claims defined in section D(2) of PR Notice
86-5, either asserting that no claims of confidentiality under
8
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FIFRA section lO(d)(1)(A), (B), or (C) are made for the study, or
that information claimed confidential under that authority has all
been removed from the body of the study to a confidential attach-
rnent. Note that these statements apply only to confidentiality
claims under FIFRA section 1O(d)(l)(A), (B), or (C); that you will
have no further opportunity to register claims of confidentiality
not made at this time; and that the wording of the two forms of the
statement is mandatory . You may add to either statement as appro-
priate, but the language specified must all be included and must
not be altered or contradicted. This statement must be signed and
dated by an authorized representative of the submitter.
C. Statement of Good Laboratory Practice Compliance. Page 3 of
each Phase 3 study must bear an appropriate statement concerning
the compliance or non-compliance of the original study with EPA ’s
Good Laboratory Practices regulations (40 CFR 160).
Note that while a statement of GLP compliance is always re-
quired, its wording is not prescribed. Since many of the studies
reformatted in Phase 3 will have been performed before these regu-
lations were in place, a statement to that effect will often be all
that is necessary or appropriate. See 40 CFR 160.12 for more
information about this requirement.
Under the regulations the GLP compliance statement must be
signed by the study director, the study sponsor, and the study
submitter. Clear, legible photocopies of appropriate statements
from the original study report are acceptable. If the study
director or study sponsor is unavailable to sign a new statement,
this should be explained in the statement, and it must be signed
by the submitter.
D. Body of the Study. This should be prepared as specified in
the Phase 3 Guidance material appropriate to the Guideline topic
addressed.
E. Confidential Attachment. The confidential attachment, if any,
and its cover sheet, as defined in section Do) of PR Notice 86-5,
follow the body of the study within the same binding.
F. Supplemental Statement of Data Confidentiality Claims. A
supplemental statement of data confidentiality claims based on
authority other than FIFRA section l0(d)(l)(A), (B), or (C), if
present, as defined in section D(4) of PR Notice 86-5, follows the
confidential attachment to the study, if any, or the body of the
study if there is no confidential attachment.
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X. Physical Format Reauirements for Phase 3 Submissions . With
the exception of a reduction in the number of copies required (see
paragraph D below), physical requirements for Phase 3 submissions
correspond to those established in PR Notice 86-5.
A. Size and Media. Each document submitted in Phase 3 must be
on uniform 8 1/2 by 11 inch white paper, printed on one side only
in black ink, with high contrast and good resolution. Be espe-
cially careful to ensure legibility of photocopied materials and
signatures. Do not submit Phase 3 materials in any media other
than paper without specific instructions from EPA.
B. Binding. Each Phase 3 summary, or each reformatted Phase 3
study, must be separately bound. Confidential attachments and
supplemental statements of data confidentiality must be included
within the binding of the body of the summary or report. All
bindings must be secure, but easily removable to permit micro-
filming.
C. Pagination. All pages of each summary or study must be con-
tinuously paginated, beginning with the required title page as page
1. The total number of pages in the body of the document must
appear on the title page and all other pages. If a confidential
attachment is present, the pagination should start over at page 1
on the cover page of the confidential attachment, and the total
number of pages in the confidential attachment must appear on each
of its pages. If a supplemental statement of data confidentiality
claims is present, it, too, must be separately paginated.
D. Number of Copies. Only one copy is required of any Phase 3
response materials . Please do not submit extra copies for any
reason.
XI. Submission of Phase 3 Materials . All Phase 3 submissions must
be addressed to:
Reregistration Phase 3 Response
US EPA: Office of Pesticide Programs
P.O. Box 14890
Silver Spring MD 20910—4890
Note that this is a USPS postal box, which cannot accept deliveries
by private courier. Please submit all Phase 3 responses via USPS;
if you need a receipt, you may send responses by certified mail,
return receipt requested. Please do not attempt to deliver Phase
3 responses to any other address. It is critical to efficient
processing that these submissions all arrive at the same place,
already separated from all the other materials directed to OPP.
10
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XII. For Further Information contact by mail:
John M. Carley
US EPA: Office of Pesticide Programs (H7504C)
401 M Street SW
Washington DC 20460
or by courier:
Document Processing Desk
Attn: John M. Carley
Crystal Mall #2, Room 266
1921 Jefferson Davis Highway South
Arlington VA 22202
or by phone:
(703) 557—2315.
c7& i
Allan S. Abramson, Director
Program Management and
Support Division
Attachment 1 Example Title Page for Phase 3 Summary
Attachment 2 Example Title Page for Reformatted Study
Attachment 3 Organization of Overall Phase 3 Submission Package
Attachment 4 Organization of Individual Summary or Reformat
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Attachn nt 1
Example Title Page for Phase 3 Summary
Phase 3 Summary of MRID 123456 and Related
MRIDs 234567, 245678, and 245679: Teratology of
Buggywhip to Albino Rabbits: Project 98765.
Guideline Reference:
83-3(b): Teratogenicity/Rabbjt
Summary Prepared by
Able B. Charles, PhD.
April 1, 1990
Original Study Prepared by:
Extraordinary Laboratories, Inc.
Kennebunkport, Maine
In Cooperation with:
Arcane Consulting Specialists, Inc.
Port Townsend, Washington
Page 1 of 17
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Attachj nt 2
Example Title Page for Reformatted Study
Phase 3 Reformat of MRID 123456 and Related
MRIDs 234567, 245678, and 245679: Teratology of
Buggywhip to Albino Rabbits: Project 98765.
Guideline Reference:
83-3(b): Teratogenicity/Rabbjt
Study Authors:
David E. Farmer
Georgeanne H. Incaviglia
James K. Lamb
Original Study Date:
March 15, 1979
Prepared by:
Extraordinary Laboratories, Inc.
Kennebunkport, Maine
In Cooperation with:
Arcane Consulting Specialists, Inc.
Port Townsend, Washington
Page 1 of 845
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Attachrr nt 3
Organization of Overall Phase 3 SUbmission Package:
1. Phase 3 Worksheet/
Transmittal Form
2. Phase 3 Summaries,
each separately bound
3. Phase 3 Reformats,
each separately bound
4. All other Phase 3
Response Materials
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Attachment 4
Organization of Individual Summary or Reformat
(All within same binding)
A. Mandatory Elements:
1. Title Page (page 1 of x)
2. Statement of Data Con-
fidentiality Claims
(page 2 of x)
3. GLP Compliance Statement
(page 3 of x)
4. Certifications of Access
to Raw Data and of
Accuracy/Adequacy
(page 4 of x)
5. Body of Summary or Study
(pages 5 of x thru x of x)
B. Optional Elements--present
only when Section lO(d)(l)
Confidentiality Claims made:
1. Cover Sheet for Confiden-
tial Attachment
(page 1 of y)
2. Body of Confidential
Attachment (pages 2 of y
thru y of y)
C. Optional Supplemental Claims
of Data Confidentiality
(pages 1 of z thru z of z)
B(1)
I I
L
I Ii
I 3(2) Ii
I I’
i
II
I C
L ..JJ
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
APPENDIX C: SPECIFIC GUIDANCE BY GUIDELINE REFERENCE NUMBER
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TABLE OF CONThNTS
Subdivision D .
Subdivision E 11
Subdivision F 83
Subdivision 129
Subdivision K 153
Subdivision L 159
Subdivision M 169
Subdivision N 279
Subdivision 0 359
Subdivision R 401
Subdivision U 407
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SUBDIVISION D
61 Product Identity and Composition 4
62 Analysis and Cenificanon of Product Ingredients 6
63 Physical and Chemical Characteristics 8
0.3
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Subdivision D
Guideline Ref. No. 61
December 24, 1989
61 Pmduct Identity and Composition
ACCEPTANCE CRiTERIA
Doos your study meet the following acceptance criteria?
1. — Name of technical material tested (include product name and trade name, if appropriate)
2. — Name, nominal concentration, and certified limits (upper and lower) for each active
ingredient and each intentionally-added inert ingredient
3. — Name and upper .fled limit for each impurity or each group of impurities present at
0.1% by weight an certain toxicologically significant impurities (e.g., dioxins,
nitrosamines) pres <0.1%
4. — Purpose of each a gredient and each intentionally-added inert
5. — Chemical name fn mical Abstracts Index of Nomenclature and Chemical Abstracts
Service (CAS) Reg “lumber for each active ingredient arid, if available, for each
intentionally-added int rt
6. — Molecular, structural, and empirical formulas, molecular weight or weight range, and any
company assigned experimental or internal code numbers for each active ingredient
7. — Description of each beginning material in the manufacturing process
— EPA Registration Number if registered; for other beginning materials, the following:
— Name and address of manufacturer or supplier
— Brand name, trade name or commercial designation
— Technical specifications or data sheets by which manufacturer or supplier describes
composition, properties or toxicity
8. — Description of manufacturing process
— Statement of whether batch or continuous process
Relative amounts of beginning materials and order in which they are added
— Description of equipment
— . Description of physical conditions (temperature, pressure, humidity) controlled in
each step and the parameters that are maintained
— Statement of whether process involves intended chemical reactions
— Flow chart with chemical equations for each intended chemical reaction
— Duration of each step of process
— Description of purification procedures
— Description of measures taken to assure quality of final product
9. Discussion of formation of impurities based on established chemical theory addressing (1)
each impurity which may be present at 0.1% or was found at ? 0.1% by product analyses
and (2) certain toxicologically significant impurities (see #3)
Criteria marked with a * are supplemental and may not be required for every study.
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Subdivision D
Guideline Ref. No. 61
December 24, 1989
61 Product Identity and Composition
GUIDANCE FOR SUMMARIZING STUDIES
The following criteria apply to the technical grade of the active ingredient being reregistered. Items 1,
2, 3, and 5 can be satisfied for most registered products by submission of the Certified Statement of
Formula Ingredients Page (EPA Form 8570-4). Items 7 and 8 can be satisfied for most technical grade
active ingredients (TGAIs) by submission of a flow chart with chemical equations for each intended
chemical reaction. The flow chart should include complete chemical structures and names for each
reactant and product of all the reactions.
Items in summaly should include the items discussed in Chapter 2 of this package and the specific items
listed below.
1. Name of technical material (include product name and trade name, if appropriate).
2. Description of each active and intentionally-added inert ingredient, including name, concentration,
and certified limits.
3. Name and upper limit for all impurities present at 0.1% and those toxicologically significant
impurities present at <0.1%.
4. The purpose of each active and intentionally-added inert ingredient.
5. Chemical name and Registry Number for each active and intentionally-added inert ingredient (if
available).
6. Molecular, structural, and empirical formulas, molecular weight, and any experimental or internal
code number for each active ingredient.
7. Description of each beginning material in the manufacturing process.
8 Description of manufacturing process.
9. Discussion of formation of Impurities based on established chemical theory.
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Subdivision D
Guideline Ref. No. 62
December 24, 1989
62 Analysis and Certifi tion of Produ Ingredients
ACCEPTANCE CRITERIA
The following criteria apply to the technical grade of the active ingredient being reregistered. Use a
table to present the information in items 6, 7, and 8.
Does your study meet the following aazptanee criteria?
1. — Five or more representative samples (batches in case of batch process) analyzed for each
active ingredient and all impurities present at 0.1%
2. — Degree of accountability or closure > ç 98%
3. — Analyses conducted for certain trace toxic impurities at lower than 0 1% (examples,
nitrosamines in the case of products containing dinitroanilines or containing secondary or
tertiary amines/alkanolamines plus nitrites; polyhalogenated dibenzodioxins and
dibenzofuraus) [ Note that in the case of nitrosamines both fresh and stored samples must be
analyzed.)
4. — Complete and detailed descnption of each step in analytical method used to analyze above
samples
5. — Statement of precision and accuracy of analytical method used to analyze above samples
6. Identities and quantities (including mean and standard deviation) provided for each analyzed
ingredient
7. Upper and lower certified limits proposed for each active ingredient and intentionally added
inert along with explanation of how the limits were determined
8 — Upper certified limit proposed for each impurity present at 0.1% and for certain
toxicologically signiflcant impurities at <0.1% along with explanation of how limit
determined
9. — Analytical methods to venfy certified limits of each active ingredient and impurities (latter
not required if exempt from requirement of tolerance or if generally recognized as safe by
FDA) are fully described
10 — Analytical methods (as discussed in #9) to verify certified limits validated as to their
precision and accuracy
Criteria marked with a * are supplemental and may not be required for every study.
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Subdivision D
Guideline Ref. No. 62
December 24, 1989
62 Anal s and Certification of Product Ingredients
GUiDANCE FOR SUMMARIZING S11JDIES
The following criteria apply to the technical grade of the active ingredient being reregistered.
Items in summary should include the items discussed in chapter 2 of this package and the specific items
listed below.
1 Number of representative samples analyzed for all active ingredients and all impurities present at >
0 1%.
2 Degree of accountability or closure in analyses in item #1.
3. Chemical names of toxic impurities which were analyzed for levels <0.1%.
4. Brief description(s) of analytical method(s) used to measure active ingredients and impurities in
items #1 and #3.
5. Statement of precision and accuracy of method(s) in item #4.
6. Chemical name and quantities observed (range, mean, standard deviation) for each ingredient
(actives and impurities) analyzed in item #1.
7. Proposed upper and lower certified limits for each active ingredient and intentionally added inert
with brief explanation of how limits were determined.
8. Proposed upper certified limit for each impurity present at >=0.1% and certain toxicologically
significant impurities at <0.1% with brief explanation of how limits were determined.
9. Brief description of analytical method(s) used to verify certified limits (if same methods as item #4,
may reference latter).
10. Statement of precision and accuracy of method(s) in item #9 (may reference item #5 if applicable).
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Subdivision D
Guideline Ref. No. 63
December 24, 1989
63 Physical and Qiemical aiaractcristica
ACCEPTANCE CRITERIA
The following criteria apply to the technical grade of the active ingredient being reregistered.
Does your study meet the following a ptance aiteria?
63-2 Color
— Verbal description of coloration (or lack of it)
— Any intentional coloration also reported in terms of Munsell color system
63-3 Physical State
Verbal description of physical state provided using terms such as “solid, granular,
volatile liquid”
— Based on visual inspection at about 20-25°C
63.4 Odor
— Verbal description of odor (or lack of it) using terms such as “garlic-like,
charactenstic of aromatic compounds”
— Observed at room temperature
63-5 Melting Point
Reported in °C
— Any observed decomposition reported
63-6 Boiling Point
— Reported in °C
Pressure under which B.?. measured reported
— Any observed decomposition reported
63-7 Density, Bulk Density, Specific Gravity
Measured at about 20-25°C
Density of technical grade active ingredient reported in glml the specific gravity of
liquids reported with reference to water at 20°C. [ Note: density of registered
products may be reported in lbs/ft 3 or lbs/gallon.]
63-8 Solubility
— Determined in distilled water and representative polar and non-polar solvents,
including those used in formulations and analytical methods for the pesticide
— Measured at about 20-25°C
Reported in g/lOOml (other units like ppm acceptable if sparingly soluble)
Criteria marked with a * are supplemental and may not be required for every study.
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Subdivision D
Guideline Ref. No 63
December 24, 1989
63-9 Vapor Pressure
— Measured at =25°C (or calculated by extrapolation from measurements made at
higher temperature if pressure too low to measure at 25°C)
— Experimental procedure described
Reported in mm Hg (torr) or other conventional units
63-10 Dissociation Constant
— Experimental method described
— Temperature of measurement specified (preferably about 20-25°C)
63-11 Octanoljwater Partition Coefficient
— Measured at about 20-25°C
— Experimentally determined and description of procedure provided (preferred method-
45 Fed. Register 77350)
— Data supporting reported value provided
63-12 pH
— Measured at about 20-25°C
— Measured following dilution or dispersion in distilled water
63-13 Stability
— Sensitivity to metal ions and metal determined
— Stability at normal and elevated temperatures
— Sensitivity to sunlight determined
Criteria marked with a * are supplemental and may not be required for every study.
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Subdivision D
Guideline Ref. No. 63
December 24, 1989
63 Physical and Chemical Characzeristics
GUIDANCE FOR SUMMARLZIJJQ FUDIES
The following criteria apply to the technical grade of the active ingredient being reregistered.
Items in sunimaiy should include the items discussed in Chapter 2 of this package and the specific items
listed below.
1. Description of color.
2. Description of physical state.
3. Description of odor.
4. Indication of melting point (in °C).
5. Indication of boiling point (in °C).
6. Indication of density, bulk density, and specific gravity.
7. Indication of solubility.
8. Indication of vapor pressure.
9. Indication of dissociation constant.
10. Indication of octanollwater partition coefficient.
11. Indication of pH.
12. Description of stability.
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SUBDIVISION E
71-1 Acute LD5O Test for Waterfowl and Upland Game Birds 12
71-2 Acute Dietary LC5O Test for Waterfowl and Upland Game Birds 17
71-3 Wild Mammal Toxicity Test . 22
71-4 Avian Reproduction Test for Waterfowl and Upland Game Birds 27
71-5 Simulated or Actual Field Testing for Birds and Mammals 33
72-1 Acute Toxicity Test for Freshwater Fish 36
72-2 Acute Toxicity Test for Freshwater Invertebrates 41
72-3 Acute Toxicity Test for Estuarine Fish 46
72-3(b) Acute Toxicity Test for Shrimp 51
72-3(c) Acute Toxicity Test for 48-hour Embryo Larvae Study and 96-hour Oyster Shell
Deposition Study 56
72-4 Early Life Stages Test . . . 61
72-4(b) Invertebrate Life-Cycle Test (Daphnia) . 66
72-4(c) Invertebrate Life-Cycle Test (Mysid) . . . 71
72-5 Fish Life-Cycle Test 76
72-7 Simulated or Actual Field Testing for Aquatic Organisms . . . 81
C-il
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Subdivision E
Guideline Ref. No. 71.1
December 24, 1989
71-1 Acute LD5O Test for Waterfowl and Upland Game Birds
ACCEPTANCE CRITERIA
The Agency will accept in fulfillment of this requirement a citation by the registrant of an appropriate
study cited in one of the publications listed below. To be considered appropnate, a study must be
conducted on the species and age of animal stipulated in the acceptance criteria, and must employ the
form of chemical designated by the acceptance criteria. If a study that appears in any of the documents
listed below meets those critena, a registrant may be exempted from summarizing the study if the
registrant submits a photocopy of the title page frorr publication and a photocopy of the
summarized study report from the publication. The ations that list studies acceptable to EPA are:
US. Department of the Intenor, Manual of Acute -- : Interpretation and Data Base for 410
Chemicals and 66 Species of Freshwater Animals ; Fi id Wildlife Service, Resource Publication 160
U.S. Department of the Interior, Handbook of Toxicity of Pesticides to Wildlife ; Fish and Wildlife
Service, Resource Publication 153
U.S. Department of the Intenor, Lethal Dietary To,ucities of Environmental Pollutants to Birds ; Fish
and Wildlife Service, Resource Publication 191
U S. EPA, Acute To cicitv Handbook of Chemicals to Estuanne Organisms : Office of Pesticides and
Toxic Substances; EPAJ600/X-86f231; September 1986.
Does your study meet the following aa ptance iteña?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERLALSIMETH0DS
2. Test birds
— Mallard duck or bobwhite quail tested
Rationale for selection of species if the species used is different than that preferred
in Subdivision E
— Age at the beginning of the test is at least 16 weeks old
‘ Method of selection of the sex of the birds descnbed and sex ratio given
3. Breeding histoiy
Histoiy of the colony, including observed diseases or other medical problems and
treatment of diseases
Indicates if all of the birds are from the same hatch, pen reared, previously mated or
unmated, phenotypically similar to wild counterpart, in a reproductive state brought
on artificially, and identifies the individuals
Criteria marked with a * are supplemental and may not be required for every study.
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Subdivision E
Guideline Ref. No. 71-1
December 24, 1989
4 Pretest penod described
_____Acclimation period (days)
*_ Pretest diet, including identification and composition plus any antibiotics, vitamins, or
food additives added to the feed
• Housing conditions, including number of animals/cage, ambient temperature and
humidity, photoperiod and lighting, and source of feed and water
• Health, including mortality, sickness, and weight loss
— Fasting period (hours)
5. Dosage described
— Amount of test material dosed/bird
— Method of administration (vehicle control/treatment), including oral incubation and
size and type of capsule, if used
*_ Rationale for selection of method, route, or frequency, if different from that
recommended in Subdivision E
6. Methods described
Number of treatment levels with a minimum of 5
— Number of birds/treatment level and control(s) (10)
Method of assigning birds to treatment levels and control groups
Method used to determine treatment levels
Size of cages, including height, width, and length in cm
Number of birds/cage
— Number of controls
Type of controls, including solvent, carrier, and whether negative or positive
7. Pens described
— Location, including whether indoors or outdoors
_ For indoor pens, ambient temperature, humidity, photoperiod, and lighting recorded
— Source and availability of feed and water identified
8. Additional factors and observations described
— Diet and any antibiotics, vitamins, or food additives added to the feed described
— Schedule and methods for gross necropsy examinations described, if performed
‘ Tissue necropsies listed, if performed
— Length of total observation period (minimum of 14 days) identified
— Frequency of each observation identified
— Bird weights reported, including when and how determined
— Food consumption data for treated and control birds reported
• Technical terminolo used to describe signs of intoxication defined
Criteria for determining effects described
Criteria marked with a * are supplemental and may not be required for every study.
C-13
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Subdivision E
Guideline Ref. No. 71.1
December 24, 1989
DISCUSSION AND RESULTS
9. Include LD5O value in mg/kg with 95% confidence limits; the slope of the dose response
line, if the method provides the slope; and the no-observed -effect level if determined. Raw
mortality data provided.
10. Description of the nature, incidence, time of occurrence, seventy, and duration of all
observed toxic effects, including death and any other abnormal or unusual signs and
symptoms. For example, report any specific signs of intoxication associated with death and
the time of onset and remission of toxic signs. Also report marked observations of fecal
material.
11. Gross pathological changes as noted by gross necropsies examination described.
12. Control bird parameters, behavior, and general health which appear atypical are described.
13. Statistical method referenced.
14. The relationship between the results of the study and the test chemical described, induding
Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
Rationale for major deviations from the protocol
15.* Graphs, printouts, and other calculations attached to the report
Criteria marked with a * are supplemental and may not be required for every study.
C- 14
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Subdivision E
Guideline Ref. No. 71-1
December 24, 1989
71-1 Acute LD5O Test for Waterfowl and Upland Game Birds
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should indude the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of test birds, including
• Species tested (scientific and common names)
- Rationale for selection of species, if species used is different than that preferred on Subdivision
E
- Age at the beginning of test for each species
- Sex ratio
3. Description of breeding history, including
- Whether the birds were from the same hatch, pen reared, and previously mated
- Identification of individuals
4 Description of the pretest period, including
- Fasting period (hours)
5. Description of dosage, including
• Method of administration (vehicle control/treatment)
- Rationale for selection of method, route, or frequency, if different from that recommended in
Subdivision E
6 Description of methods, including
- Number of treatment levels
- Number of birds/treatment level and control(s)
- Number of birds/cage
- Number of controls
- Type of controls, including solvent, carrier, and whether negative or positive
7. Description of pens, including
- Location, including whether indoors or outdoors
8. Description of additional factors and observations, including
- Length of total observation period (number of days)
C-15
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Subdivision E
Guideline Ref. No. 71-1
December 24, 1989
DISCUSSION AND RESULTS
9. Results, including
• Statistical method referenced
- LD5O value (mg/kg) with 95% confidence limits
• The no-observed-effect level, if determined
- Bird weights reported, including when determined
- Food consumption data for treated and control birds
10. Discussion relating the results of the study to test chemical, including
• An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
- The rationale for any major deviations from recommended protocol.
C-16
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Subdivision E
Guideline Ref. No. 71-2
December 24, 1989
71-2 Acute Dictazy LC5O Test for Waterfowl and Upland Game Birds
ACCEPTANCE CRITERIA
The Agency will accept in fulfillment of this requirement a citation by the registrant of an appropriate
study cited in one of the publications listed below. To be considered appropriate, a study must be
conducted on the species and age of animal stipulated in the acceptance criteria, and must employ the
form of chemical designated by the acceptance criteria. If a study that appears in any of the documents
listed below meets those criteria, a registrant may be exempted from summanzing the study if the
registrant submits a photocopy of the title page from the publication and a photocopy of the
summarized study report from the publication. The publications that list studies acceptable to EPA areS
U S. Department of the Interior, Manual of Acute Toxicity: Interpretation and Data Base for 410
Chemicals and 66 Species of Freshwater Animals : Fish and Wildlife Service, Resource Publication 160
U.S. Department of the Interior, Handbook of Toxicity of Pesticides to Wildlife ; Fish and Wildlife
Service, Resource Publication 153
U S. Department of the Interior, Lethal Dietary Toxicities of Environmental Pollutants to Birds ; Fish
and Wildlife Service, Resource Publication 191
U.S. EPA, Acute Toxicity Handbook of Chemicals to Estuanne Organisms ; Office of Pesticides and
Toxic Substances; EPA/600/X-86,231; September 1986.
Does your study meet the following a ptance criteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test diets (control/treatment) described
Any vehicle used to dissolve or dilute test substance with the feed
— Amount of vehicle added to control diet
_ Description of test diet and any other ingredients such as antibiotici, vitamins, or
food additives which may have been in the feed preceding or during testing
°_ Type of mixer and method of mixing in the test material and vehicle
Report data if food was analyzed for the pesticide concentration
Method used to determine concentration levels
Criteria marked with a e are supplemental and may not be required for every study.
C-17
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Subdivision E
Guideline Ref. No. 71-2
December 24, 1989
3. Test birds
— Mallard duck and bobwhite quail tested
— Rationale for selection of species if the species used is different than that preferred
in Subdivision E
— Age at the beginning of the test is 5 to 10 days old for mallard ducks and 10 to 14
days for bobwhite quail
4. Breeding histoty
— Birds were phenotypically similar to wild counterpart, all from the same hatch, and
pen reared.
— Any observed diseases and treatments were reported
5. Descnption of any pretest conditioning
* Health
• Description of pretest diet, including any antibiotks, vitamins, or food additives
_ Housing conditions described, including number of animals/cage, ambient temperature
and humidity, photopenod and lighting, and source of feed and water
6. Pen facilities described
Dimensions of test pen(s) in cm
— Temperature (C°) and relative humidity
— Photoperiod and lighting
Pen locations (indoors and outdoors)
* Pen construction materials
Availability of water throughout the test
_ Provisions for ventilation
7. Concentration and dosage mortality/effects data
Method of identifying treatment levels
— Descnption of the steps taken to maintain the concentration(s), if the test substance
is administered in the feed, and if over the length of the test concentrations may
decrease by 30% or more
— Number of concentrations (5 or 6)
— Number of days on test matenal (5)
Number of days on food free of test matenal (3)
Number of birds/cage
— Number of birds/concentration and control group(s) (10)
— Method for assigning test organisms to test and control groups (random)
8.*_ Methods for determining bird body weights or other physiological measurements described
9 s Food consumption - colleclion methods
Criteria marked with a * are supplemental and may not be required for eveiy study.
C-18
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Subdivision E
Guideline Ref. No. 71-2
December 24, 1989
10. Observations
— Frequency of observations
— Signs of intoxication and abnormal behavior
— Palatability or repellency
_ List tissue necropsied, if performed
‘ Cnteria for determining effects
_ Define technical terminolo ’ used to describe signs of intoxication
DISCUSSION AND RESULTS
11 — Include LC5O value in ppm with 95% confidence limits; the slope of the dose response line,
if the method provides the slope; and the no-observed-effect level if determined. Raw
mortality data provided.
12.*_ Gross pathological changes as noted by gross necropsies examination, including
characterization of gross lesions
13. Description of the nature, incidence, time of occurrence, severity, and duration of all
observed toxic effects, including death and any other abnormal or unusual signs and
symptoms. For example, report any specific signs of intoxication associated with death and
the time of onset and remission of toxic signs Also report marked observations of fecal
material.
14. — Statistical method referenced.
15. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for major deviations from the protocol
16.’_ Graphs, pnntouts, and other calculations attached to the report.
Criteria marked with a * are supplemental and may not be required for every study.
C-19
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Subdivision E
Guideline Ref. No. 71-2
December 24, 1989
71-2 Acute Dietary LC5O T t for Waterfowl and Upland Game Birds
GUIDANCE FOR SUMMARIZING STUDIES
Items in summaiy should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of lest diet (control/treatment) preparation, including
- Any vehicle used to dissolve or dilute test substance with the feed
3 Description of test birds, including
- Species tested (scientific and common names)
- Rationale for selection of species, if species used is different than that preferred on Subdivision E
- Age at the beginning of test for each species
4. Description of breeding history, including
- Whether the birds are from the same hatch and pen reared
5. Description of any pretest conditioning
6. Description of pen facilities, including
- Pen locations (indoors and outdoors)
7. Description of concentration and dosage monaliiy/effects data, including
- Number of concentrations
- Number of days on test material
- Number of days on food free of test material
- Number of birds/cage
- Number of birds/concentration and control group(s)
8. Description of observations, including
- Frequency of observations
C-20
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Subdivision E
Guideline Ref No. 71-2
December 24, 1989
DISCUSSION AND RESULTS
9 Results, including
- Statistical method referenced
- LCSO value (ppm) with 95% confidence limits
- The no-observed-effect level if determined
- Signs of intoxication and abnormal behavior
- Palatability or repellency
10 Discussion relating the results of the study to test chemical, including
- An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
- The rationale for any major deviations from recommended protocol.
C-21
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Subdivision E
Guideline Ref. No. 71-3
Detember 24, 1989
71-3 Wild Mamm il Toxicity Test
ACCEPTANCE CRiTERIA
Does your study meet the following aa ptance criteria?
GENERAL INFORMATION
1 Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test material (control/treatment) described
— The vehicle used to dissolve test material
— Amount of vehicle added to the control
Method used to get test material into diet
— Total amount of test material used
3 Test mammal
— Rationale for selection of species if the species used is different than those exposed
to test substance’s use pattern
— Test species common and scientific name
_ History of test organisms (strain, disease, and treatment)
4. Pretest conditioning described
— Health, including sickness, Injuries, abnormalities, name of medication (if used), and
pretest diet
Quarantine, including number of days mammals held
5 Size/age/physical condition described
— Age of each animal and how determined
— Size of the animal, including the weight of any animal more than ± 20% of mean
weight
— Physical condition of test animals
Critena marked with a * are supplemental and may not be required for every study.
C-22
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Subdivision E
Guideline Ref. No. 71-3
December 24, 1989
6. Acclimation
‘__ Daily food consumption
_ Daily behavior of test animal
— Body weights, including at the beginning and end of acclimation
Diet used
— Source of animal food and water
— Dimensions of pen(s)
_ Length of photoperiod
7 Physical characteristics of testing facility described
— Identification of any special laboratory equipment unique to the study
8. Biological characteristics of testing facility descnbed
— Daily temperature measurements
*_ Daily relative humidity measurements
— Daily photopertod
‘ Daily laboratory air exchange rate
9. Range finding test as the test exposure parameter
— Number of animals used
Method used to determine dose levels
— Raw data for determining the end point
— Treatment and observation period
— Any statistical analysis utilized
10 Definitive test
— Number of animals used and a rationale for using less than 10 animals per treatment
level, if appropriate
— Th4umber of treatment levels
— Name of vehicle used
*_ References to toxicity of vehicle
— Controls and vehicle controls used
Raw mortality or effects data for each group
— Duration of treatment and observation periods
— Statistical analysis of the data
— Methodology for dosing
Methodology for treatment diet preparation
Toxicologically related behavioral change
Analysis of basal diet
— Criteria for end point of study
— Number of animals of each sex of animal tested
11. _Gross necropsies conducted
Criteria marked with a * are supplemental and may not be required for every study.
C-23
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Subdivision E
Guideline Ref. No. 71-3
December 24, 1989
DISCUSSION AND RESULTS
12. Body weight and food consumption
*_ Daily body weight by animal and level
* Total daily food consumption by animal and level
— Mean body weight for each test and control group at initiation and termination of
test
13. — LC5O value in ppm with 95% confidence limits; the slope of the dose response line, if the
method provides the slope; and the no-observed-effect level, if determined. Raw mortality
data provided.
14 — Description of the nature, incidence, time of occurrence, severity, and duration of all
observed toxic effects, including death and any other abnormal or unusual signs and
symptoms. For example, report any specific signs of intoxication associated with death and
the time of onset and remission of toxic signs. Also report marked observations of fecal
material and urine.
15. — Gross pathological changes as noted by gross necropsies examination described.
16. Control mammal response, behavior, and general health.
17 — Statistical method referenced.
18 The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study.
19 *_ Graphs, printouts, and other calculations attached to the report
Criteria marked with a * are supplemental and may not be required for every study.
C-24
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Subdivision E
Guideline Ref. No. 71-3
December 24, 1989
71-3 Wild Mammal Toxicity Test
GUiDANCE FOR SUMMARIZING STUDIES
[ tems in summaly should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of test material (control/treatment) preparation, including
- The vehicle used to dissolve test material
3. Description of test mammal, including
- Rationale for selection of species if the species used is different than those exposed to test
substance’s use pattern
- Species tested (common and scientific names)
- History of test organisms (strain, disease, and treatment)
4 Description of pretest conditioning, including
- Health, including sickness, injuries, abnormalities, name of medication (if used), and pretest diet
- Quarantine, including number of days mammals held
5 Descripuon of size/age/physical condition, including
- Age of each animal and how determined
- Size of the animal, including the weight of any animal more than ± 20% of mean weight
- Physical condition of test animals
6. Description of acclimation, including
- Diet used
7 Description of physical characteristics of testing facility, including
• Identification of any special laboratory equipment unique to the study
C.25
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Subdivision E
Guideline Ref. No. 71.3
December 24, 1989
8. Descnption of definitive test, including
- Number of animals used and a rationale for using less than 10 animals per treatment level, if
appropriate
- Number of treatment levels
- Name of vehicle used
• References to toxicity of vehicle
- Controls and vehicle controls used
• Statistical analysis of the data
- Methodolo ’ for dosing
• Toxicologically related behavioral change
• Number of animals of each sex of animal tested
9. Description of gross necropsies conducted
DISCUSSION AND RESULTS
10. Results, including
- Statistical method referenced
• LC5O value (ppm) with 95% confidence limits
• The no-observed-effect level, if detennined
- Gross pathological changes as noted by gross necropsies examination
• Control mammal response, behavior, and general health.
- Mean body weight and food consumption for each test and control group at initiation and
termination of test
ii. Discussion relating the results of the study to test chemical, including
An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
C.26
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Subdivision E
Guideline Ref. No. 71-4
December 24, 1989
71-4 Avian Reproduction Test for Waterfowl and Upland Game Birds
ACCEPTANCE CRITERIA
Does your study meet the following acceptance criteria?
GENERAL INFORMATION
1 Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test diets (control/treatment) described
— Any vehicle used to dissolve or dilute test substance with the feed
— Amount of vehicle added to control diet
— Description of test diet and any other ingredients such as antibiotics, vitamins, or
food additives which may have been in the feed preceding or during testing
— Type of mixer and method of mixing in the test material and vehicle
*_ Report data if food was analyzed for the pesticide concentration
— Method used to determine concentration levels
3 Test birds
— Mallard duck and bobwhite quail tested
— Rationale for selection of species if the species used is different than that preferred
in Subdivision E
4 Breeding history
— Birds were phenotypically similar to wild counterpart, pen reared, and were in the
first breeding season.
My observed diseases and treatments were reported
5. Description of any pretest conditioning
* Health
‘_ Description of diet, including identification and composition
Housing conditions described, including number of animals/cage, ambient temperature
and humidity, photoperiod and lighting, and source of feed and water
_ Length of observation
Criteria marked with a * are supplemental and may not be required for every study.
C-27
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Subdivision E
Guideline Ref. No. 71-4
December 24, 1989
6. Concentration and dosage mortality data
— Number of concentrations
— Number of days on test material including time prior to egg laying
— Number of animals/cage
— Randomization plan for treatments
— Method of identifying treatment levels
Length of observation period (including recovery period, if necessary)
— Description of the steps taken to maintain the concentration(s), if the test substance
is administered in the feed or water
— Method of assigning test organisms to test and control groups
7. Experimental design
A minimum of 3 test groups, one control and 2 treatment groups
— For mallards, 2 males and 5 females/pen, replicated by .2: pens/trtatment group; or
if individual pairs are used/pen, 2:12 pens/treatment group are required
For bobwhite quail, one male and 2 females/pen, replicated by .2:12 pens/treatment
group; or if individual pairs are used/pen, >12 pens/treatment group are required
8. Methods for measuring bird body weights or other physiological measures described
9. Food consumption
— Collection methods
* Describe collection schedule
* Provisions for minimizing food spillage and preventing air contamination by volatile
chemicals
10. Pen facilities described
— Pen size and construction, including space allocations for mating and nesting;
protection from weather if outdoors; and protection from injuries
— Photopenod and lighting, including hours/day and wattage or footcandles at bird level
— Temperature and humidity
* Pen location
. Diagram of test layout
_ Ventilation of pen area
— Source and availability of water
— Pen construction materials
Criteria marked with a * are supplemental and may not be required for every study.
C-28
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Subdivision E
Guideline Ref. No. 71-4
December 24, 1989
11. Egg collection, storage, and incubation
Date egg laying began reported
Egg collecting, cleaning, and storage procedures described
Days eggs candled for eggshell cracks, embryo viability, and embryo survival reported
— Egg rotation regime described
Procedure used in selecting eggs for egg shell measurements reported
Day eggs placed in hatcher and day removed reported
Method of identifying each bird described
Schedule and method for gross necropsies examinations, if performed
— Incubator temperature and humidity
— Hatching egg storage temperature and humidity
12. — Describe grouping of hatchlings in relation to treatment levels and parents
13 — Tissue residue data from extra pen group if the test chemical is known to bioaccumulate, if
performed
14. Define technical terminology used to describe signs of intoxication
DISCUSSION AND RESULTS
15 — A summary of reproductive data and reproductive success data.
16 Reproductive data for controls and treatment levels
Eggs laid
— Eggs cracked
— Eggs set
— Viable embryos
— Live three-week embryos
— Normal hatchlings
— 14 day-old survivors
17. Reproductive success data for controls and treatment levels
— Eggs laid/ben in 8 weeks
— Eggs cracked of eggs laid (%)
Viable embryos of eggs set
Live three-week embryos of viable embryos (%)
Normal hatchling of live three-week embryos
14 day-old survivors of normal hatchlings
— 14 day-old survivors/ben
Criteria marked with a * are supplemental and may not be required for every study.
C-29
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Subdivision E
Guideline Ref. No. 71-4
December 24, 1989
18. Reproductive data for controls and treatment levels
— Eggshell thickness data, including number of eggs analyzed and mean shell thickness
— Weight (grams) of 14-day old survivors for each bird
— Body weight and feed consumption data for each bird for duration of the study
— Reproduction data for each pen, including eggs laid, eggs cracked, eggs set, viable
embryos, live 3-week embryos, normal hatchlings, and egg shell thickness data
— 14-day old survivors by week, for at least 8 weeks
19 _ If any statistical differences are found discuss their biological significance
20. Statistical method(s) used for analyzing data are referenced.
21 Describe and discuss control bird reproduction parameters behavior, and general health which
appear atypical.
22. — Description of nature, incidence, time ol occurrence, severity, and duration of all observed
toxic effects, including all morphological and physiological responses, death and any other
abnormal or unusual signs and symptoms For example, report any specific signs of
intoxication associated with death and the time of onset and remission of toxic signs. Also,
report recovery time for affected birds In addition, marked observations of fecal material
should be reported.
23. Gross pathological changes as noted by observation or gross necropsies examination including
characterization of gross lesions
24 Repori repellency of the diet or lack of palatability.
25 * No-observed-effect level, if determined
26. — Report the results of residue analysis of test diet
27.*_ Degree of bioaccumulation if this portion of the study is performed
28. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred dunng the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for major deviations from the protocol
29 * Graphs, printouts and other calculations are attached to the report
Cnteria marked with a * are supplemental and may not be required for every study.
C-30
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Subdivision E
Guideline Ref. No 71-4
December 24, 1989
71-4 Avian Reproduction Test for Watcrfowl and Upland Game Birds
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of test diet (control/treatment) preparation, including
- Any vehicle used to dissolve or dilute test substance with the feed
3. Description of test birds, including
- Species tested (common and scientific names)
- Rationale for selection of species, if the species used is different than that preferred in
Subdivision E
4. Description of breeding history, including
- Similarity (in terms of phenotype) of birds to wild counterpart
- Whether the birds were pen reared and in the first breeding season
5. Description of any pretest conditioning, Including
- Health
6. Description of concentration and dosage mortality data, including
- Number of concentrations
- Number of days on test material
- Number of animals/cage
- Length of observation period
7. Description of experimental design, including
- Number of test groups, control, and treatment groups
- For each species, number of birds (males and females)/pen, replication of pens/treatment group;
or if individual pairs are used/pen, number of pens/treatment
8. Description of pen facilities, including
- Pen size and construction, including space allocations for mating and nesting; protection from
weather if outdoors; and protection from injuries
C-31
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Subdivision E
Guideline Ref. No. 71.4
December 24, 1989
9. Description of egg collection, storage, and incubation
- Date egg laying began
- Method of identifying each bird
10. Tissue residue data from extra pen group if the test chemical is known to bioaccumulate, if
performed
DISCUSSION AND RESULTS
11. Description of the reproductive data for controls and treatment levels, including
- Eggs laid
- Eggs cracked
- Eggs set
- Viable embryos
- Live three-week embryos
- Normal hatchlings
- 14 day-old survivors
12 Description of the reproductive success data for controls and treatment levels, per dose
- Eggs laid/hen in 8 weeks
- Eggs cracked of eggs laid (%)
- Viable embryos of eggs set
Live three-week embryos of viable embryos (%)
- Normal hatchling of live three-week embryos
- 14 day-old survivors of normal hatchlings
- Mean value of 14 day-old survivors/hen
13. Description of the reproductive data for controls and treatment levels, per dose
• Mean value of eggshell thickness (mm)
- Mean value of weight (grams) of 14-day old survivors for each bird
- 14-day old survivors by week, for at least 8 weeks
14. Discussion of biological significance, if any statistical differences are found, including references for
statistical method(s) used for analyzing data are referenced.
15 Repellency of the diet or lack of palatability reported.
16. Determination of the no-observed-effect level.
17. Results of residue analysis of test diet reported.
18. Degree of bioaccumulation, if this portion of the study is performed.
19. Discussion relating the results of the study to test chemical, including
- An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
- The rationale for any major deviations from recommended protocol.
C -32
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Subdivision E
Guideline Ref. No. 71-4
December 24, 1989
71-5 Simulated or Actual Field Testing for Birds and Mammals
ACCEPTANCE CRITERIA
Does your study meet the following aceeptance criteria?
1.’_ The protocol was submitted/reviewed by the Agency.
2.*_ The following document was reviewed when registrant considered the suitability of
summarizing the study and relying on it to support reregistratton
Guidance Document for conducting Terrestrial Field Studies, EPA 540/09-88-109, September
1988. NTIS No. PB89-124580.
This reference is available from the National Technical Information Service (NTIS). Orders may be
placed to NTIS by telephone at (703) 487-4650 or by mail to the following address:
National Technical Information Service
A1TN. Order Desk
5285 Port Royal Road
Springfield, Virginia 22161
Criteria marked with a * are supplemental and may not be required for every study.
C-33
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Subdivision E
Guideline Ref. No. 71-5
December 24, 1989
71-5 Simulated or Actual Field Testing for Birds and Mamm ’Lc
GUIDANCE FOR SUMMARIZING STUDIES
Items in sumniaty should include the items discussed in Chapter 2 of this package and the specific items
listed below.
1. INTRODUCTION
- Objective of Field 5
- General Approach
- Sampling and Experimental Design
2. SCREENING STUDY
- Objective and Scope
- Geographic Area Selection
- Study Site Selection
- Number of Sites
- Size of Study Sites
- Chemical Application
- Methods
- Carcass Searches
- Radio Telemetry
- Test of Cholinesterase Inhibition
• Residue Anz jysis
- Behavioral Observations
- Density and Diversity Estimates
- Interpretation of Results
C-34
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3 DEFINITIVE STUDY
- Objective and Scope
- Sampling and Experimental Design
- Study Area and Site Selection
- Number and Size of Sites
- Methods
- Mortality and Survival
- Mark-Recapture
- Territory Mapping Method
- Radio Telemetry
- Other Methods for Mortality and Survival
- Reproduction and Survival of Dependent Young
- Nest Monitoring
- Behavioral Observations
• Age Structure of Populations
- Ancillary Methods
- Interpretation of Results
4. LITERATURE CITED
C-35
Subdivision E
Guideline Ref. No. 71-5
December 24, 1989
-------
Subdivision E
Guideline Ref. No. 72-1
December 24, 1989
fl-i Acute Toxicity Test for Freshwater Fish
ACCEPTANCE CRITERIA
The Agency will accept in fulfillment of this requirement a citation by the registrant of an appropriate
study cited in one of the publications listed below. To be considered appropriate, a study must be
conducted on the species and age of animal stipulated in the acceptance cnteria, and must employ the
form of chemical designated by the acceptance criteria. If a study that appears in any of the documents
listed below meets those criteria, a registrant may be exempted from summarizing the study if the
registrant submits a photocopy of the title page from the publication and a photocopy of the
summarized study report from the publication. The publications that list studies acceptable to EPA are:
U.S. Department of the Interior, Manual of Acute Toxicity: Interpretation and Data Base for 410
Chemicals and 66 Species of Freshwater Animals ; Fish and Wildlife Service, Resource Publication 160
U.S. Department of the Interior, Handbook of Toxicity of Pesticides to Wildlife ; FISh and Wildlife
Service, Resource Publication 153
U.S. Department of the Intenor, Lethal Dietary Toxicities of Environmental Pollutants to Birds ; Fish
and Wildlife Service, Resource Publication 191
U.S. EPA, Acute Toxicity Handbook of Chemicals to Estuarine Organisms ; Office of Pesticides and
Toxic Substances; EPA/600/X-861231; September 1986.
Does your study meet the following a p1ance criteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test solution (control/treatment) descnbed
— Any vehicle used to dissolve test matenal
If used, amount of vehicle added to control
Method used to get test material into solution was described
Water source
— Water solubility of test material reported in ppm
*_ Total amount of test material used
_ Temperature of water
Criteria marked with a * are supplemental and may not be required for every study.
C-36
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Subdivision E
Guideline Ret No. 72-1
December 24, 1989
3 Test fish
— One of the following organisms was tested: rainbow trout, bluegill, brook trout,
coho salmon, channel catfish, fathead minnow
History of test organisms described (strain, diseases, and treatment)
‘_ Health described including sickness, injunes, abnormalities, name of medication (if
used), and pretest diet
_ Age
— Weight in grams (0.5 to 5) and length in mm reported
4. Source/acclimation described -
Complete name and address of test fish supplier, source of food and dilution water, size of
test container in liters (L), temperature, pH, dissolved oxygen, name of equipment used to
measure water quality, feeding schedule, holding and acclimation period, test organisms from
the same source, number of days held.
5. Composition and size of test vessels described
— Material type
Volume in liters (L)
— Depth of test solution in cm or volume of dilution water (L)
6. Test system described
— Source of dilution water
— If flow-through, system and flow rate/day described
— Procedures used to prepare toxicant stock solution
— Criteria used to determine effects
7 Test design followed
_ Method used in assigning test organisms to test and control groups and the number
of replicates used
— Number of fish per dose level and control group (minimum 10 for static)
__ Name of protocol followed dunng this test
Loading (weight per unit volume of water with no more than 0.8g/L for static and
1.0 g/L for flow-through)
— 5 treatments and 1 control used
— Length of exposure period (96-hours minimum)
— Type of control (positive, negative, or solvent control)
Temperature (12±2°C for coldwater fish and 17±2°C or 22±20 C for warmwater
fish)
_pH
•• Lighting (time in hours and intensity - footcandles)
— Dissolved oxygen
* Water hardness expressed in mgfL as CaCO 3
* Alkalinity expressed in mgIL as CaCO 3 )
* Conductivity (umhos/cm)
‘_ Range finding test results, concentrations, and mortality, if used
Criteria marked with a * are supplemental and may not be required for every study.
C-37
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Subdivision E
Guideline Ref. No. 72.1
December 24, 1989
Water not aerated during exposure to pesticides (unless concentrations are measured)
“. .. Water: physical characteristic at the end of test - depth and volume
DISCUSSION AND RESULTS
8. — LC5O va’ue reported in ppm with 95% confidence limits. Raw mortality data provided.
9. — Any precipitation and solubility problems.
10. — Statistical method referenced.
ii. D sion of concentration(s) producing intoxication provided.
Water: physical characteristic at end of test - temperature, pH, and dissolved oxygen
Concentration analysis (if flow through)
_ Fish physiology, locomotton, behavior and pathology described
— Mortalities during the test described
12.*_ No-observed-effect level, if determined.
13. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for minor deviations from the protocol
14.*.._. Graphs, pnntouts and other calculations are attached to the report
Criteria marked with a are supplemental and may not be required for every study.
C-38
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Subdivision E
Guideline Ref. No 72-1
December 24, 1989
fl-i Acute Toxicity Tcst for Fresh ter Fish
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2 Description of the test solution (control/treatment) preparation, including
- Any vehicle used to dissolve test material
- Water solubility of test material (ppm)
3. Description of test species (fish), including
- Organism(s) tested (common and scientilic names)
- Weight (g) and length (mm)
4 Description of source/acclimation, including
• Complete name and address of test fish supplier
5 Description of composition and size of test vessels, including
- Material type
6 Description of test system, including
• System and flow rate/day, if flow-through
7. Description of test design, including
• Number of organisms/dose level and control group
• Name of protocol followed during test
• Loading
- Number of treatments and control
- Length of exposure period
- Type of control
- Temperature
-pH
- Dissolved oxygen
- Water aeration during exposure to pesticides
C-39
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Subdivision E
Guideline Ref. No. 72-1
December 24, 1989
DISCUSSION AND RESULTS
8. Results, including
- Statistical method referenced
- LC5O value (ppm) with 95% confidence limits
- Description of any precipitation and solubility problems
- Discussion of concentration(s) producing intoxication
- The no-observed-effect level, if determined
- Concentration analysis (if flow through)
- Mortalities during the test
9. Discussion relating the results of the study to test chemical, including
- An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study
- The rationale for any major deviations from recommended protocol.
C-40
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Subdivision E
Guideline Ref. No. 72-2
December 24, 1989
72-2 Acute Toxicity Test for Freshwater Invertebrates
ACCEPTANCE CRITERIA
Does your study meet the following acceptance aitcna?
GENERAL INFORMATION
Date of test initiation and termination reported
MATERIALS/METHODS
2 Preparation of test solution (control/treatment) described
— Any vehicle used to dissolve test material
— If used, amount of vehicle added to control
— Method used to get test material into solution was described
Water source
— Water solubility of test material reported in ppm
* Total amount of test material used
Temperature of water reported
3. Test invertebrate
— One of the following organisms was tested: Daphnia magna, Daphnia pulex ,
mayflies, amphipods, stoneflies
_ History of test organisms described (strain, diseases and treatment)
* Health described including sickness, injuries, abnormalities, name of medication (if
used), pretest diet
— Daphnids were in first instar ( . 24 hours old) and other invertebrates were in the
second instar.
4. Source/acclimation described -
complete name and address of supplier, source of food and dilution water, size of test
container in milliliters (mL), temperature, pH, dissolved oxygen, name of equipment used to
measure water quality, feeding schedule, holding, acclimation period, test organisms are from
same source, and number of days adults held.
5. Composition and size of test vessels described
— Material type
— Volume in liters (L) or size in centimeters (cm)
Depth of test solution (cm) or volume of dilution water (L)
Criteria marked with a * are supplemental and may not be required for every study.
C-4 I
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Subdivision E
Guideline Ref. No. 72-2
December 24, 1989
6 Test system descnbed
— Source of dilution water
— If flow-through, system and flow rate/day described
Procedures used to prepare toxicant stock solution
Critena used to determine effects
7. Test design followed
— Method used in assigning test organisms to test and control groups and number of
replicates used
— At least 20 Daphnia per dose level and control group
_ Name of protocol followed during this test
— 5 treatments and 1 control used
— Length of exposure period (48 hours for Daphnids and 96 hours for other
invertebrates)
— Type of control (positive, negative, or solvent control)
— Temperature (20±2°C for Daphnids , 17±2°C for amphipods and mayflies, and
12±2°C for stoneflies)
_pH
Lighting (time in hours and intensity - footcandles)
— Dissolved oxygen
— Water hardness expressed in mg/L as CaCO 3
_ Alkalinity expressed in mgfL as CaCO 3
*_ Conductivity (umhos/cm)
* Range flnding test results (concentrations and mortality), if conducted
— Water not aerated during exposure to pesticide (unless concentrations are measured)
_ Water: physical characteristic at the end of test - depth and volume
DISCUSSION AND RESULTS
8 — LCSO or EC5O value reported in ppm with 95% confidence limits. Raw mortality data
provided.
9 — Any precipitation and solubility problems described.
10. — Statistical method referenced.
11. — Discussion of concentration(s) producing intoxication provided
— Water: physical characteristic at the end of test - temperature, pH, and dissolved
oxygen
— Concentration analysis (if flow through)
— Invertebrate behavior described
— Mortalities during test described
12.* No-observed-effect level, if determined
Criteria marked with a are supplemental and may not be required for every study.
C-42
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Subdivision E
Guideline Ref. No. 72-2
December 24, 1989
13. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the Study
— Rationale for minor deviations from the protocol
14 *_ Graphs, printouts and other calculations are attached to the report
Criteria marked with a * are supplemental and may not be required for every study.
C-43
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Subdivision E
Guideline Ref. No. 72.2
December 24, 1989
fl-2 Acute Toxicity Test for Freshwater Invertebrates
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2 Description of the test solution (control/treatment), preparation including
- Any vehicle used to dissolve test material
- Water solubility of test material (ppm)
3. Description of test species, including
- Organism(s) tested (common and scientific names)
- Stage
4. Description of source/acclimation, including
- Complete name and address of supplier
5. Description of composition and size of test vessels, including
- Material type
6 Description of test system, including
- System and flow rate/day, if flow-through
7 Description of test design, including
- Number of organisms/dose level and control group
- Name of protocol followed during lest
• Number of treatments and control
- Length of exposure period
- Type of control
- Temperature
-p1-I
- Dissolved oxygen
- Water aeration during exposure to pesticides
C-44
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Subdivision E
Guideline Ref. No. 72-2
December 24, 1989
DISCUSSION AND RESULTS
8. Results, including
- Statistical method referenced
- LCSO or EC5O value (ppm) with 95% confidence limits
- Description of any precipitation and solubility problems
- Discussion of concentration(s) producing intoxication
- The no-observed-effect level (if determined)
- Concentration analysis (if flow through)
- Invertebrate behavior
- Mortalities during the test
9 Discussion relating the results of the study to test chemical, including
• An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
- The rationale for any major deviations from recommended protocol.
C-45
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Subdivision E
Guideline Ref. No. 72-3
December 24, 1989
72-3 Acute Toxicity Test for Estuarine Fish
ACCEPTANCE CRiTERIA
The Agency will accept in fulfillment of this requirement a duation by the registrant of an appropriate
study cited in one of the publications listed below. To be considered appropriate, a study must be
conducted on the species and age of animal stipulated in the acceptance criteria, and must employ the
form of chemical designated by the acceptance criteria. If a study that appears in any of the documents
listed below meets those criteria, a registrant may be exempted from summarizing the study if the
registrant submits a photocopy of the title page from the publication and a photocopy of the
summanzed study report from the publication. The publications that list studies acceptable to EPA are
U.S. Department of the Interior, Manual of Acute Toxicity: Interpretation and Data Base for 410
Chemicals and 66 Species of Freshwater Animals ; Fish and Wildlife Service, Resource Publication 160
U.S. Department of the Interior, Handbook of Toxicity of Pesticides to Wildlife ; Fish and Wildlife
Service, Resource Publication 153
U.S. Department of the Interior, Lethal Dietary Toxicities of Environmental Pollutants to Birds ; Fish
and Wildlife Service, Resource Publication 191
U.S EPA, Acute Toxicity Handbook of Chemicals to Estuarine Organisms ; Office of Pesticides and
Toxic Substances; EPA/600(X-86,231; September 1986.
Does your study meet the following a ptance aiteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test solution (controlltreatment) described
Any vehicle used to dissolve test material
— If used, amount of vehicle added to control
Method used to get test material into solution was described
* Water source
— Water solubility of test material reported in ppm
Total amount of test material used
• Temperature of water reported
* Salinity (o/oo)
Criteria marked with a * are supplemental and may not be required for every study.
C-46
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Subdivision E
Guideline Ref. No. 72.3
December 24, 1989
3. Test fish
— One of the following organisms was tested: silverside and sheepshead minnow
‘ History of test organisms described (strain, diseases and treatment)
_ Health described (sickness; injuries; abnormalities; name of medication, if used;
pretest diet, number of days brood fish were quarantined)
_ Age, if known
— Weight ranges from 0.5 to 5 grams and length reported (mm)
4 — Source/acclimation descnbed.
complete name and address of test fish supplier, source of food and dilution water, size of
test container in liters (L), temperature, pH, dissolved oxygen, name of equipment used to
measure water quality, feeding schedule, holding, acclimation period, test organisms are from
same source, salinity (o/oo)
5. Composition and size of test vessels described
— Material type
— Volume in liters (L) or size in centimeters (cm)
— Depth of test solution in cm or volume of dilution water
6. Test system described
— Source of dilution water
— If flow-through, system and flow rate/day described
— Procedures used to prepare toxicant stock solution
— Criteria used to determine effects
7 Test design followed
— Method used in assigning test organisms to test and control groups and number of
replicates used
— Minimum of 10 fish for static and 20 fish for flow-through per dose level and control
group
_ Name of protocol followed during this test
— Loading (weight per unit volume of water with no more than 0.8 g/L for static and
1.0 g/L for flow-through)
— 5 treatments and 1 control used
Minimum of 96 hour exposure period
l’ype of control (positive, negative, or solvent control)
— Temperature (22±2° C)
_pH
_ Ughting (time in hours and intensity - footcandles)
— Dissolved oxygen
‘ Conductivity (umhos/cm)
_ Range finding test results (concentrations and mortality) if conducted
— Water not aerated during exposure to pesticide (unless concentrations are measured)
_ Water: physical characteristic at the end of test - depth and volume
Cntena marked with a * are supplemental and may not be required for every study.
C-47
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Subdivision E
Guideline Ref. No. 72-3
December 24, 1989
DISCUSSION AND RESULTS
8. LC5O value reported in ppm with 95% confidence limits. Raw mortality data provided.
9. Any precipitation and solubility problems described.
10. — Statistical method referenced.
11. Discussion of concentration(s) producing intoxication provided
— Water: physical characteristic at the end of test - temperature, pH, and dissolved
oxygen
— Concentration analysis (if flow through)
— Fish physiology, locomotion, behavior, and pathology described
— Mortalities during test
12.* No-observed-effect level, if determined
13. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for minor deviations from the protocol
14.*_ Graphs, printouts and other concentrations should be attached to the report
Critena marked with a * are supplemental and may not be required for every study.
C -48
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Subdivision E
Guideline Ref. No. 72-3
December 24, 1989
72-3 Acute Toxicity Test for Estuarine Fish
GUIDANCE FOR SUMMARIZING STUDIES
Items in summpry should include the itcms discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test solution (control/treatment), preparation, including
- Any vehicle used to dissolve test material
- Water solubility of test material (ppm)
3. Description of test species (fish), including
- Organism(s) tested (common and scientific names)
- Weight (g) and length (mm)
4 Description of source/acclimation, including
- Complete name and address of test fish supplier
5 Description of composition and size of test vessels, including
- Material type
6. Description of test system, including
- Source of dilution water
- System and flow rate/day, if flow-through
7. Description of test design, including
- Number of lish for static and for flow-through/dose level and control group
- Name of protocol followed during test
- Loading
- Number of treatments and control
- Length of exposure penod
- Type of control
- Temperature
-pH
- Dissolved oxygen
- Water aeration during exposure to pesticides
C-49
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Subdivision E
Guideline Ref. No. 72-3
December 24, 1989
DISCUSSION AND RESULTS
8. Results, including
- Statistical methods referenced
- LC5O value (ppm) wflh 95% confidence limits
- Descnption of any precipitation and solubility problems
• Discussion of concentration(s) producing intoxication
• The no-observed-effect level if determined
- Concentration analysis (if flow-through)
- Fish physiology, locomotion, behavior, and pathology described
- mortalities during test
9. Discussion relating the results of the study to test chemical, including
- An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study
- The rationale for any major deviations from recommended protocol.
C-50
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Subdivision E
Guideline Ref. No. 72-3(b)
December 24, 1989
72-3(b) Acute Toxicity Test for Shrimp
ACCEPTANCE CRITERIA
Does vur study meet the following acceptance criteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2 Preparation of test solution (control/treatment) described
— Any vehicle used to dissolve test material
— If used, amount of vehicle added to control
— Method used to get test material into solution was described
Water source
— Water solubility of test material reported in ppm
* Total amount of test material used
* Temperature
_ Salinity (0/00)
3 Test shrimp
One of the following organisms was tested: white, pink, brown, grass and m sid
*_ History of test organisms described (strain, diseases, and treatment)
‘_ Health described, including sickness, injuries, abnormalities, name of medication (if
used), and pretest diet
— Juvenile stage was tested, except for mysids which must be . 24 hrs old
— Weight measured in grams and length in mm, except for mysids
4 — Source/acclimation described -
Complete name and address of test shrimp supplier, source of food and dilution water, size
of test container in liters (L), temperature, pH, dissolved oxygen, name of equipment used to
measure water quality, feeding schedule, holding and acclimation period, test organisms from
the same source, number of days held, salinity (o/oo).
5. Composition and size of test vessels described
— Material type
— Volume in liters (L) or size in centimeters (cm)
— Depth of test solution in cm or volume of dilution water (L)
Criteria marked with a * are supplemental and may not be required for every study.
C-5 1
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Subdivision E
Guideline Ref. No. 72.3(b)
December 24, 1989
6. Test system described
— Source of dilution water
— If flow-through, system and flow rate/day described
Procedures used to prepare toxicant stock solution
Cnteria used to determine effects
7. Test design followed
Method used in assigning test shrimp to test and control groups and the number of
replicates used
Number of shrimp per dose level and control group (minimum 10 for static and 20
for flow-through)
_ Name of protocol followed during this test
Loading (weight per unit volume of water with no more than 0.8 g/L for static and
1.0 g/L for flow-through)
— 5 treatments and I control
Length of exposure period (96-hours minimum)
— Type of control (positive, negative, or solvent control)
— Temperature (22±2° C)
_pH
* Lighting (me in hours and intensity - footcandles)
— Dissolved oxygen
*_ Range finding test results, concentrations, and mortality, if used
— Water not aerated dunng exposure to pesticides (unless concentrations are measured)
_ Water: physical characteristic at the end of test - depth and volume
DISCUSSION AND RESULTS
8. — LC5O value reported in ppm with 95% confidence limits. Raw mortality data provided.
9. — Any precipitation and solubility problems.
10. — Statistical method referenced.
11. — Discussion of concentration(s) producing intoxication provided.
— Water: physical characteristic at end of test - temperature, pH, dissolved oxygen, and
salinity (o/oo)
— Concentration analysis (if flow-through)
— Shrimp physiology, locomotion, behavior and pathology described
— Mortalities during the test described
12.’ No-observed-effect level, if determined.
Criteria marked with a * are supplemental and may not be required for every study.
C-52
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Subdivision E
Guideline Ref. No. 72.3(b)
December 24, 1989
13. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
Rationale for minor deviations from the protocol
14, Graphs, printouts and other calculations are attached to the report
Criteria marked with a are supplemental and may not be required for every study.
C .53
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Subdivision E
Guideline Ref. No. 72-3(b)
December 24, 1989
72-3(b) Acute Toxicity Test for Shrimp
GUiDANCE FOR SUMMARIZING STUDIES
[ tems in summary should include the itcms discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test solution (control/treatment), preparation, including
- Any vehicle used to dissolve test material
- Water solubility of test material (ppm)
3 Description of test species, including
- Organism(s) tested (common and scientific names)
- Stage
- Weight (g) and length (mm), except for mysids
4. Description of source/acclimation, including
- Complete name and address of test shrimp supplier
- Holding and acclimation period
5. Description of composition and size of test vessels, including
- Material type
6. Description of test system, including
- Source of dilution water
- System and flow rate/day, if [ low-through
7. Description of test design, including
- Number of shrimp/dose level and control group
- Name of protocol followed during test
- Loading
- Number of treatments and control
- Length of exposure period
- Type of control
- Temperature
-pH
- Dissolved oxygen
- Water aeration during exposure to pesticides
- Salinity (0/00)
C-54
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Subdivision E
Guideline ReL No. 72.3(b)
December 24, 1989
DISCUSSION AND RESULTS
8. Results, including
- Statistical method referenced
- LC5O value (ppm) with 95% confidence limits
- Descnpuon of any precipitation and solubility problems
- Discussion of concentration(s) producing intoxication
The no-observed-effect level if determined
- Concentration analysis, if flow-through
9 Discussion relating the results of the study to test chemical, including
- An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
The rationale for any major deviations from recommended protocol.
C-55
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Subdivision E
Guideline Ref. No. 72.3(c)
December 24, 1989
fl-3(c) Acute Toxicity Test for 4&hour Embryo Laivac Study
and 96-hour O3 ster Shell Deposition Study
ACCEPTANCE CRITERIA
Does your study meet the followmg acceptance criteria?
GENERAL INFORMATION
Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test solution (control/treatment) described
— Any vehicle used to dissolve test material
— If used, amount of vehicle added to control
— Method used to get test material into solution was described
Water source
— Water solubility of test material reported in ppm
Total amount of test material used
_ Salinity (0/00)
3 Test oyster
— Atlantic oyster tested
History of test organisms described (strain, diseases, and treatment)
— Describe fertilization process and method used to collect embryos
— For the shell deposition study, oysters should be 25.50 mm in height and for the
oyster embryo larvae study, embryos should be 1.hour old)
_ List abnormalities, if any occurred
4. — Source/acclimation described -
Complete name and address of test species supplier, source of food and dilution water, size
of test container in liters (L), temperature, pH, dissolved oxygen, name of equipment used to
measure water quality, Feeding schedule, holding and acclimation period, test organisms from
the same source, salinity (0/00).
5. Composition and size of test vessels described
— Material type
— Volume in liters (L)
Depth of test solution in cm or volume of dilution water (L)
Criteria marked with a * are supplemental and may not be required for every study.
C -56
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Subdivision E
Guideline Ref. No. 72-3(c)
December 24, 1989
6. Test system described
— Source of dilution water
— If flow-through, system and flow rate/day described
— Procedures used to prepare toxicant stock solution
— Criteria used to determine effects
7. Test design followed for the “embryo study”
— Method used in assigning test organisms to test and control groups
— Number of fertilized eggs/L of test solution (20,000-30,000)
— 5 treatments and 1 control used
Length of treatment penod (48 hours)
— Type of control (positive, negative, or solvent control)
_ Temperature prior to and during spawning
_pH
*_ Lighting (time in hours and intensity - footcandles and transition period between
light and dark)
— Dissolved oxygen
‘_ Range finding test results, if used
— Water not aerated during exposure to pesticides (unless measurements are taken)
* Water: physical characteristic at the end of rest depth and volume
_ How embryos were collected and preserved
8 Test design followed for the “shell deposition study”
— Method used in assigning test organisms to test and control groups
— Number of organisms per dose level (20)
— 5 treatments and 1 control
— Length of exposure period (96 hours)
— Type of control (positive, negative, or solvent control)
_pH
Lighting (time in hours and intensity - footcandles and transition period between
light and dark)
— Dissolved oxygen
Range finding test results, if used
Water: physical characteristic at the end of test - depth and volume
•_ Name of protocol used during the test
“_ How peripheral valve edge was removed
— Method used to measure oyster size
DISCUSSION AND RESULTS
9. — EC5O value reported in ppm with 95% confidence limits. Raw mortality data provided.
10. — Any precipitation and solubility problems.
11. — Statistical method referenced.
Criteria marked with a * are supplemental and may not be required for every study.
C-57
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Subdivision E
Guideline Ref. No. 72-3(c)
December 24, 1989
12. — Discussion of concentration(s) producing intoxication provided.
— Water: physical characteristic at end of test - temperature, pH, dissolved oxygen, and
salinity (oloo)
— Number of normal developed larvae at the end of observation
— Number of deformed larvae at the end of observation
— Percent death/effects at each dose level
— Concentration analysis (if flow through)
— Length of new shell growth
13.*_ No-observed-effect level, if determined.
14. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for minor deviations from the protocol
15.*_ Graphs, printouts and other calculations are attached to the report
Criteria marked with a * are supplemental and may not be required for every study.
C-58
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Subdivision E
Guideline Ref No. 72-3(c)
December 24, 1989
72-3(c) Acute Toxicity Test for 48-hour Embryo Larvae Study
and 96-hour Oyster Shell Deposition Study
GUIDANCE FOR SUMMARIZING STUDIES
Items in summaiy should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test solution (control/treatment), preparation, including
- Any vehicle used to dissolve test material
- Water solubility of test material (ppm)
3. Description of test species, including
- Organism(s) tested (common and scientific names)
- For shell deposition study, height of oysters and for the oyster embryo larvae study, age of
embryos (hocrs)
4 Description of source/acclimation, including
- Complete name and address of test species supplier
5 Description of composition and size of test vessels, including
- Material type
6. Description of test system, including
- Source of dilution water
- System and flow rate/day, if flow-through
7. Descnption of test design for the “embryo study’, including
- Number of fertilized eggsfL of test solution
- Number of treatments and controls
- Length of treatment period (hours)
- Type of control
- Name of protocol used during test
- Dissolved oxygen
- Water aeration during exposure to pesticides
C-59
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Subdivision E
Guideline Ref. No. 72-3(c)
December 24, 1989
8. Description of test design for the sheIl deposition study , including
- Number of organisms/dose level
- Number of treatments and controls
- Length of exposure penod (hours)
- Type of control
- Dissolved oxygen
- Water aeration during exposure to pesticides
- Name of protocol used during the test
DISCUSSION AND RESULTS
9. Results, including
- Statistical method referenced
- ECSO value (ppm) with 95% confidence limits
- Descnption of any precipitation and solubility problems
- Discussion of concentration(s) producing intoxication
- No-observed-effect level, if determined
- Length of new shell growth
- Number of normal developed larvae at the end of observation
- Number of deformed larvae at the end of observation
10. Discussion relating the results of the study to test chemical, including
- An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
• The rationale for any major deviations from recommended protocol.
C-60
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Subdivision E
Guideline Ref. No. 72-4
December 24, 1989
72-4 Early Life Stages Test
ACCEPTANCE CRITERIA
Does your study meet the following acceptance criteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2 Preparation of test solution (control/treatment) described
Any vehicle used to dissolve test material
If used, amount of vehicle added to control
Method used to get test material into solution was described
Water source
— Water solubility of test material reported in ppm
_ Total amount of test material used
_ Temperature of water reported
* Salinity (0/00), if cstuarine species is tested
3. Test fish
One of the following organisms was tested: brook trout, rainbow trout, echo salmon.
chinook, bluegill, brown trout, lake trout, northern pike, fathead minnow, channel
catfish, sheepshcad minnow, silverside
_ History of test organisms described (strain, diseases and treatment)
_ Health described (sickness; injuries; abnormalities; name of medication, if used,
pretest diet, number of days brood fish were quarantined)
4 Size/age/physical condition
_ Age of unfertilized eggs and milt at the beginning of test
* Procedures used to strip eggs and milt described
*_ Procedures used in storage and handling of eggs described
— Fertilization process described
5* Source/acclimation described -
Complete name and address of test fish supplier, source of food and dilution water, size of
test container in milliliters (mL), temperature, pH, dissolved oxygen, name of equipment used
to measure water quality, feeding schedule, holding, acclimation period, test organisms are
from same source, number of days held, salinity (o/oo)
Criteria marked with a * are supplemental and may not be required for every study.
C-61
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Subdivision E
Guideline Ref. No. 72-4
December 24, 1989
6. Composition and size of test vessels descnbed
— Material type
— Volume in liters (L) or size in centimeters (cm)
— Depth of test solution in cm or volume of dilution water (L)
7. Test system described
— Source of dilution water
— If flow-through, system and flow rate/day described
— Procedures used to prepare toxicant stock solution
— Criteria used to determine effects
8. Test design followed
— Method used in assigning test organisms to test and control groups and number of
replicates used
Number of eggs per dose level and control group (80)
__ Name of protocol followed during this test
— Loading (weight per unit volume of water)
5 treatments and I control used
Length of exposure period
— Type of control (positive, negative, or solvent control)
— Temperature
* Lighting (time in hours and intensity - footcandles)
— Dissolved oxygen
Water hardness
* Alkalinity
_ Conductivity (umhos/cm)
. .__ Range finding test results (concentrations and mortality) if conducted
* Water: physical characteristic at the end of test - water depth and volume
— Period food was withheld prior to termination
Method used to keep eggs clean described
DISCUSSION AND RESULTS
9 MATC value reported in ppm. Raw data provided.
10. — Any precipitation and solubility problems described.
11. Statistical method referenced.
12. Discussion of concentration(s) producing intoxication provided.
— Water: physical characteristic at the end of test - temperature, pH, and dissolved
oxygen
Concentration analysis (if flow through)
Criteria marked with a * are supplemental and may not be required for every Study.
C-62
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Subdivision E
Guideline Ref. No. 72-4
December 24, 1989
13 — Signs of intoxication noted.
— Number of embryos hatched per replicate reported
— Time to hatch per replicate reported
— Mortality of embryos, larvae, and juveniles per replicate reported
— Time to swim-up per replicate reported
— Measurements of growth-weight and length per replicate reported
— Fish physiology, locomotion, behavior and pathology reported
— When larvae fish released from the retaining cup, % complete or hours elapsed
identified
14.*_ No-observed-effect level, if determined.
15. The relationship between the results of the study and the test chemical described, including
Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for minor deviations from the protocol
16.*..... Graphs, printouts and other calculations are attached to report
Criteria marked with a are supplemental and may not be required for every study.
C-63
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Subdivision E
Guideline Ret No. 72-4
December 24, 1989
72-4 Earty Life Stage Test
GUIDANCE FOR SUMMARIZING STUDIES
Items in summpry should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1 Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test solution (control/treatment), preparation, including
- Any vehicle used to dissolve test material
- Water solubility of test material (ppm)
3. Description of test species (fish), including
- Organism(s) tested (common and scientific names)
4 Description of size/age/physical condition
- Age of unfertilized eggs and milt at the beginning of test
5. Description of source/acclimation, including
- Complete name and address of test fish supplier
- Number of males and females used/batch of eggs
6. Description of composition and size of test vessels, including
- Material type
7 Description of test system, including
- Source of dilution water
- System and flow rate/day, if flow-through
- Critena used to determine effects
C-64
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Subdivision E
Guideline Ref. No. 72-4
December 24, 1989
8. Description of test design, including
- Number of eggs/dose level and control group
- Name of protocol followed during test
• Loading
• Number of treatments and control
- Length of exposure period
- Type of control
• Temperature
-pH
- Lighting (time in hours and intensity - footcandles)
- Dissolved oxygen
- Water aeration during exposure to pesticides
DISCUSSION AND RESULTS
9. Results, including
- Statistical method referenced
- Number of embryos hatched/dose
- Time to hatch/dose
- Mortality of embryos and larvae/dose
• Time to swim-up/dose
- Measurements of growth-weight and length/dose
- Percent complete or hours elapsed when larvae fish released from the retaining cup/dose
- MATC value (ppm)
- Description of any precipitation and solubility problems
- Signs of intcxication
• Determination of the no-observed-effect level
- Concentration analysis
10. Discussion relating the results of the study to test chemical, including
- An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
- The rationale for any major deviations from recommended protocol.
C-65
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Subdivision E
Guideline Ref. No. 72-4(b)
December 24, 1989
72-4(b) Lnvcncbrate Life-Cycle Test (Daphnia)
ACCEPTANCE CRITEREA
Does your study meet the following acceptance criteria?
GENERAL INFORMATION
I — Date of test initiation and termination reported
MATERIALS/METHODS
2 Preparation of test solution (control/treatment) described
— Any vehicle used to dissolve test material
— If used, amount of vehicle added to control
— Method used to get test material into solution was described
* Water source
— Water solubility of test material reported in ppm
_ Total amount of test material used
* Temperature of water reported
3 Test Daphnia
D phnia manna was tested
_ History of test organisms described (strain, diseases and treatmcnt)
* Health described (sickness; injuries, abnormalities; name of medication, if used,
pretest diet, number of days brood daphnids were quarantined)
4. Size/age/physical condition
— Age of initiation of test (124 hours)
Date spawned
* List any abnormalities reported
5. — Source/acclimation described -
Complete name and address of Daphnia supplier, source of food and dilution water, size of
test container in milliliters (mL), temperature, pH, dissolved oxygen, name of equipment used
to measure water quality, feeding schedule, holding, acclimation period, test organisms are
from same source, number of days adults held in laboratory prior to use of offspring
6. Composition and size of test vessels described
— Material type
— Volume in liters (L) or size in centimeters (cm)
— Depth of test solution in cm or volume of dilution water (L)
Criteria marked with a * are supplemental arid may not be required for every study.
C-66
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Subdivision E
Guideline Ref. No. 72-4(b)
December 24, 1989
7 Test system described
— Source of dilution water
— If renewal, schedule described
— If flow-through, system and flow rate/day descnbed
— Procedures used to prepare toxicant stock solution
— Criteria used to determine effects
8. Test design followed
— Method used in assigning test organisms to test and control groups and number of
replicates used
— Number of daphnids per dose level and control group (minimum of 22)
_ Name of protocol followed during this test
— 5 treatments and 1 control used
— Length of exposure period (21 days)
— Type of control (positive, negative, or solvent control)
— Temperature (22 ± 2° C)
_pH
— Lighting (time in hours and intensity footcandles)
— Dissolved oxygen
Water hardness
‘_ Alkalinity
‘___ Conductivity (umhos/cm)
• Range finding test results (concentrations and mortality) if conducted
* Water not aerated during exposure to pesticide (unless concentrations are measured)
* Water: physical characteristic at the end of test - water depth and volume
— Feeding regime (food, frequency, and amount)
DISCUSSION AND RESULTS
9. — MATC value reported in ppm. Raw data provided.
10. — Any precipitation and solubility problems described.
11. — Statistical method referenced.
12. — Signs of intoxication (temporal onset, duration and concentrations that showed effects) noted
— Water: physical characteristic at the end of test - temperature, pH, and dissolved
oxygen
Concentration analysis (ii flow through or aeration is supplied)
Survival of first generation daphnids reported
— Total young produced reported
— Mortality during the test reported
— Success of controls - the study is rejected if 30% total mortality, . 40 young were
produced, or if ephippia are produced
— Daphnia physiology, locomotion, behavior and pathology defined
Criteria marked with a * are supplemental and may not be required for every study.
C-67
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Subdivision E
Guideline Ref. blo. 72-4(b)
December 24, 1989
13. — The no-observed-effect level and reproductive effects defined.
14. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for major deviations from the protocol
15.*_ Graphs, printouts and other calculations are attached to the report
Criteria marked with a * are supplemental and may not be required for every study.
C -68
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Subdivision E
Guideline Ref. No. 72.4(b)
December 24, 1989
72-4(b) Envcrtcbrate Life-qrclc Test (Daphnia)
GUIDANCE FOR SUMMARIZING S11JDLES
Items in summaly should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1 Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test solution (control/treatment), preparation, including
- Any vehicle used to dissolve test material
- Amount of vehicle added to control, if used
• Water solubility of test material (ppm)
3. Description of test species, including
- Organism(s) tested (common and scientific names)
4 Description of size/age/physical condition
- Age at initiation of test
Date spawned
5 Description of source/acclimation, including
• Complete name and address of test species supplier
6. Description of composition and size of test vessels, including
- Material type
7. Description of test system, including
• System and flow rate/day, if flow-through
8. Description of test design, including
Number of organisms/dose level and control group
- Name of protocol followed during test
- Number of treatments and controls
- Length of exposure period
- Type of control
- Temperature
-p ’- 1
Dissolved oxygen
- Water aeration during exposure to pesticides
C.69
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Subdivision E
Guideline Ref. No. 72-4(b)
December 24, 1989
DISCUSSION AND RESULTS
9. Results, including
- Statistical method referenced
- MATC value (ppm)
- Description of any precipitation and solubility problems
• Determination of the no-observed-effect level and reproductive effects
concentration analysis (if flow through)
- Survwal of first generation organisms/dose
- Total young produced/dose
- Length of first generation organisms/dose
- Mortalities during test/dose
• Success of controls
10 Discussion relating the results of the study to test chemical, including
• An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
- The rationale for any major deviations from recommended protocol.
C-70
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Subdivision E
Guideline Ref. No. 72 .4(c)
December 24, 1989
fl.4(c) Invertebrate Life-Cycle Test (M id)
ACCEPTANCE CRITERIA
Does your study meet the following a ptance criteria?
GENERAL INFORMATION
1. Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test solution (control/treatment) described
— Any vehicle used to dissolve test material
— If used, amount of vehicle added to control
Method used to get test material into solution
Water source
Water solubility of test material reported in ppm
• Total amount of test material used
* Water temperature
* Salinity (o/oo)
3 Test mysid
Mvsidopsis hahia tested.
History of test organisms described (strain, diseases, and treatment)
Health described, including sickness, injuries, abnormalities, name of medication (if
used), pretest diet, number of days brood mysid quarantined
4. Size/age/physical condition
— Age of initiation of test ( 24 hours)
— Date spawned
List of any abnormalities reported
5. — Source/acclimation described.
Complete name and address of mysid supplier, source of food and dilution water, size of test
container in milliliters (mL). temperature, pH, dissolved oxygen, name of equipment used to
measw’e water quality, feeding schedule, holding and acclimation period, test organisms from
the me source, number of days adults held in laboratory prior to use of oft pring
6. Composition and size of test vessels described
— Material type
— Volume or size in liters (L)
— Depth of test solution in centimeters (cm)
Criteria marked with a • are supplemental and may not be required for every study.
C i i
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7. Test system described
— Source of dilution water
— If renewaL schedule described
— If flow-through, system and flow rate/day described
— Procedures used to prepare toxicant stock solution
Criteria used to determine effects
Subdivision E
Guideline Ref. No. 72-4(c)
December 24, 1989
8. Test design followed
— Method used in assigning test organisms to test and control groups, and the number
of replicates used
— Number of mysidsldose level and control group at the start of exposure (at least 60
orgarnsms)
— Name of protocol followed during this test
— 5 treatments and I control used
— Number of pairs for treatment and control (at least 20)
— Length of exposure period with a minimum of7 days after median time of first
brood release in control
— Type of control (positive, negative, or solvent control)
— Temperature
— pH
— Lighting (time in hours and intensity - fooicandles)
— Dissolved oxygen
R inge finding test results, concentrations, and mortality, if used
_ Water not aerated during exposure to pesticides (unless concentrations are measured)
‘ Water: physicaL characteristic at end of test - water depth and volume
— Feeding regime - food, frequency. and amount
— Examination of mysid physiolo i, locomotion, behavior, and pathoto
DISCUSSION AND RESULTS
9. Test endpoints
by replicate
Water: physical characteristic at end of test - temperature, pH, and dissolved oxygen
Concentration analysis (if flow through or aeration is supplied)
Survival of first generation mysids
Live young produced by each pair
Length and dry weight of surviving first generation mysids
Mortalities dunng the test
Success of controls; study is rejected if .30% total adult mortality between pairing
and end of test and average 3 young produced per female or 25% moriality of
females
10. — MATC value reported in ppm. Raw data provided
11, — Any precipitation and solubility problems described.
Criteria marked with a • are supplemental and may not be required for every study.
C-72
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Subdivision E
Guideline Ref. No. 7 2 -4(c)
December 24, 1989
12. — Statistical method referenced.
13. — Signs of intoxication (temporal onset, duration, and concentrations that showed effects)
noted.
14. The no-observed-effect level and reproductive effects defined.
15. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
Rationale for major deviations from the protocol
16. Graphs, pnntouts and other calculations are attached
Cntena marked with a • are supplemental and may not be required for every study.
C-73
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Subdivision E
Guideline Ref. No. 2.4(c)
December 24, 1989
72-4(c) Tnvertcbrate Life-Cycle Test (M id)
GUiDANCE FOR SUMMARIZING STUDIES
Items in sulrnnary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1 Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the preparation of test solution (control/treatment), including
Any vehicle used to dissolve test material
- Water source
• Water solubility of test material (ppm)
- Salinity (oloo)
3 Description of test species, including
• Organism(s) wsred (common and scientific names)
4. Description of size/age/physical condition
- Age of initiation of test
5 Description of source/acclimation, including
• Complete name and address of test species supplier
6. Description of composition and size of test vessels, including
- Material type
7. Description of test system, including
• Source of dilution water
- Schedule described, if renewal
- System and flow rate/day, if flow-through
C-74
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Subdivision E
Guideline Ref. No. 72-4(c)
December 24, 1989
8. Description of test design, including
- Number of replicates used
• Number of organisms/dose level and control group at start of exposure
- Name of protocol followed dunng test
- Number of treatments and controls
- Length of exposure period
• Type of control
- Temperature
-pH
- Lighting (time in hours and intensity - footcandles)
- Dissolved oxygen
- Water aeration dunng exposure to pesticides
DISCUSSION AND RESULTS
9. Description of endpoints by dose
- Concentration analysis
- Survival of first generation organisms
• Mean value of live young produced by each pair
- Mean value of length and dry weight of surviving first generation organisms
- Mortalities during test
- Success of controls
10. Results, including
- Statistical method referenced
- MATC value (ppm)
• Descnption of any precipitation and solubility problems
• Discussion of signs of intoxication (temporal onset, duration, and concentrations that showed
effects)
• Determination of the no-observed-effect level and reproductive effects
11 Discussion relating the results of the study to test chemical, including
• An explanation of obvious problems that occurred during the study and what effect, if any, these
problems had on the outcome of the study.
- The rationale for any major deviations from recommended protocol.
C-75
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Subdivision E
Guideline Ref. No. 72.5
December 24, 1989
72-5 Fish Life-Cycle Test
ACCEPTANCE CRITERIA
Does your study meet the following aceeptance criteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATER IALSIMETHODS
2. Preparation of test solution (control/treatment) described
— Any vehicle used to dissolve test material
— If used, amount of vehicle added to control
— Method used to get test material into solution was described
Water source
— Water solubility of test material reported in ppm
‘ Total amount of test material used
‘_ Temperature
Salinity (0/00), if estuarine species is tested
3 Test fish
— Fathead or sheepshead minnow lesied
‘_ History of test organisms described (strain, diseases, and treatment)
_ Health described including sickness, injuries, abnormalities, name of medication (if
used), pretest diet, number of days brood fish were quarantined
4. Size/age/physical condition
Age of unfertilized eggs and mi i i (in hours) at the beginning of test
— Procedures used to strip eggs and milt descnbed
— Procedures used ia storage and handling of eggs described
— Fertilization process described
5. — Source/acclimation described.
Complete name and address of test fish/egg supplier, source of food and dilution water, size
of test oontainer (in milliliters (rnL)j, temperature, p 1- I, and dissolved oxygen, and name of
equipment used to measure water quality, feeding schedule, holding, and acclimation period,
test organisms from the same source, number of days held, salinity (0/00), number of males
and females used per batch of eggs, percent mortality of brood fish, description of methods
used initially to determine different stages of development.
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-76
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Subdivision E
Guideline Ref. No. 72-5
December 24, 1989
6. Composition and size of test vessels described
— Material type
Volume in liters (L) or size in centimeters (cm)
Depth of test solution in cm or volume of dilution water (L)
7. Test system described
— Source of dilution water
If flow-through, system and flow rate/day described
Procedures used to prepare toxicant stock solution
— Criteria used to determine effects
8. Test design followed
— Method used in assigning test organisms to test and control groups, and the number
of replicates used
Number of eggs per dose level and control group (100)
‘ Name of protocol followed during this test
5 treatments and I control
— Length of exposure period
— Type of control (positive, negative, or solvent control)
Temperature
— pH
‘__ Lighting (time in hours and intensity - footcandles)
Dissolved oxygen
Water hardness
S Alkalinity
_ Conductivity (umhos/cm)
• Range finding test results (concentrations and mortality), if conducted
- Water: physical characteristic at end of test - water depth and volume
— Method used to keep eggs clean described
— Period food was withheld prior to termination
DESCUSSION AND RESULTS
9 Effects on 1st and 2nd generation embryo/cup reported
— Days to complete hatching
— Total number of embryos hatched
— Number of normal larvae hatched
10. Effects on 1st and 2nd generation larval.juvenile/replicate 4 to 8 weeks after hatching reported
— Number of survivors
— Number of abnormal fish
— 1)pes of abnormalities
— Total length of all survivors
Weight of all survivors
Criteria marked with a • are supplemental and may not be required for eveiy study.
C-77
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Subdivision E
Guideline Ref. No. 72-5
December 24, 1989
11 — Effects on 1st generation juvenhle adu11s/rep1icate tank reported, including total length,
weight, and sex of all fish (20 xo 24 Weeks)
12. Effects on 1st generation adult/pair reported
Total length and weight at time of pairing
Number of spawns
Number of embryos/spawn
Total length, weight, and sex of parental fish after last spawn
13. MATC value reported in ppm. Raw data provided.
14. — Any precipitation and solubility problems described.
15. — Statistical method referenced.
16. Discussion of concentration(s) producing intoxication provided.
Water: physical and chemical characteristic at end of test - temperature, pH,
dissolved oxygen and chemical
— Concentration analysis
17. Signs of intoxication noted.
Fish physiology, locomotion, behavior and pathology reported
18. — The no-observed-effect level defined.
19. — Detailed record of spawning, fertility, and fecundity exists.
When larvae fish released from the retaining cup, percent complete or hours elapsed
identified
— Time to swim-up per replicate reported
20 The relationship between the results of the study and the test chemical described, including
— Obvious problems chat occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for major deviations from the protocol
21. Graphs, printouts and other calculations are attached to report
Criteria marked with a • are supplemental and may not be required for every study.
C-78
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Subdivision E
Guideline Ref. No. 72-5
December 24, 1989
72-5 Fish tAle-Cycle Test
GUiDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
Listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALSIMETHODS
2. Description of the test solution (control/treatment), preparation, including
- Any vehicle used to dissolve test material
- Water solubility of test material (ppm)
- Salinity (0100), if estuarine species is tested
3. Description of test species (fish), including
- Organism(s) tested (common and scientific names)
4. Description of size/age/physical condition
- Age of unfertilized eggs and rnilt (in hours) at ihe beginning of test
5. Description of source/acclimation, including
- Complete name and address of test fish/egg supplier
- Source of test organisms
- Number of males and females usedThatch of eggs
- Methods used to initially determine different stages of development
6. Description of composition and size of test vessels, including
- Material type
7. Description of test system, including
- Source of dilution water
- System and flow rate/day, if flow-through
8. Descnption of test design, including
- Number of repLicates
- Number of eggs/dose level and control group
- Name of protocol followed during test
- Number of treatments and control
- Length of exposure period
- Type of control
- Temperature
C-79
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Subdivision E
Guideline Ref. No. 72-5
December 24, 1989
-pH
- Dissolved oxygen
- Water aeration during exposure to pesticides
DISCUSSION AND RESULTS
9. Descnption of test design (continued)
• When larvae fish released from the retaining cup, percent complete or hours elapsed
- Time to swim-up/replicate
10. Description of effects on 1st and 2nd generation embryo/dose, including
- Mean value of days to complete hatching
- Mean value of total number of embryos hatched
- Mean value of number of normal larvae hatched
11. Descnption of effects on 1st and 2nd generation larval-juvenile/dose 4 to 8 weeks after hatching,
including
- Number of survivors
- Number of abnormal fish
- Mean value of total length of all survivors
• Mean value of weight of all survivors
12. Description of the effects on 1st generation juvenile-adults/dose, including total length and weight
13 Description of effects on 1st generation adult/dose
- Mean value of total length and weight at time of pairing
- Number of spawns
• Number of embryos
- Mean value of total length, weight, and sex of parental fish after last spawn
14. Results, including
- Statistical method referenced
- MATC value (ppm)
- Description of any precipitation and solubility problems
- Determination of the no-observed-effect level
- Concentration analysis
15. Discussion relating the results of the study to test chemical, including
• An explanation of obvious problems that occurred during the study, as well as the effect, if any,
these problems had on the outcome of the study
• The rationale for major deviations from the protocol
C-80
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Subdivision E
Guideline Ref. No. 72-7
December 24, 1989
72-7 Simulated or Actual Field Testing for Aquatic Organisms
ACCEPTANCE CRITERIA
Generally, tests are unacceptable if conducted prior to 1986.
Does your study meet the following aocepian criteria?
BASIC CRITERIA
1. The protocol was submitted/reviewed by the Agency.
2. The following document was reviewed by the registrant when considering the suitability of
summarizing the study and relying on it to support reregistration:
Hazard Evaluation Division Technical Guidance Document - Aquatic Mesocosm Tests to
Support Pesticide Registrations, EPA 540/09.88-035, March 1988. NTIS No. PB8S-1724.65.
This reference is available from the National Technical Information Service (NTIS). Orders may be
placed to NTIS by telephone at (703) 487-4650 or by mail to the following address:
National Technical Information Service
ATTN Order Desk
5285 Port Royal Road
Springfield, Virginia 22161
Criteria marked with a * are supplemental and may not be required for every study.
C-81
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Subdivision E
Guideline Ref. No. 72.7
December 24, 1989
72-7 Simulated or Actual Field Testing for Aquatic Organisms
GUIDANCE FOR SUMMARIZING FUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the sp ific items
listed below.
1. OBJECTIvES
2. PROPOSE!) DESIGN CRITERIA
- Physical Description
• Experimental design
- Mesocosm number
- Mesocosm size
- Mesocosm features
- Mesocosm biota
- Mesocosm treatment
3. MEASURED PARAMETERS
- Chemical/Physical Properties
• Biological Structure
- Residue Analysis
Meteorological Conditions
4. RATIONALE
5. INTERPRETATION OF RESULTS
6. LITERATURE CITED
C -82
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SUBDIVISION F
81-1 Acute Oral Toxicity in the Rat .
81-2 Acute Dennal Toxicity in the Rat, Rabbit or Guinea Pig
81-3 Acute Inhalation Toxicity in the Rat
81-4 Primary Eye Irritation in the Rabbit
81-5 Primary Dermal irritation Study
81-6 Dermal Sensitization in the Guinea Pig
81-7 Acute Neurotoxicity in the I-len
82-1 Subchronic Feeding in the Rodent and Nonrodent
82-2 Repeated Dose Dermal Toxicity (21-day) in the Rat, Rabbit or Guinea
82-3 Repeated Dose Dermal Toxicity (90-day) in the Rat, Rabbit or Guinea
82-4 Subchronic Inhalation Toxicity (90-day) in the Rat
82-5 Subchronic Neurotoxicity (90-day) in the Hen
83-1 Chronic Feeding in the Rodent and Nonrodent
83-2 Oncogenicity in Rats or Mice
83-3 Teratology Studies
83-4 Reproduction
83-5 Chronic Feeding/Oncogenicity in the Rat . . 2.
84-2 Mutagenicity Studies .
85-1 Metabolism Studies
84
86
90
92
96
98
Pig 101
Pig . 103
106
109
111
114
117
119
. 121
124
127
C-83
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Subdivision F
Guideline Ref. No. 81.1
December 24, 1989
81-1 A te Oral T ty in the Rat
ACCEPTMIICE CRrrERIA
Do your study meet the Ibilowing a ptanee aiteria?
1. — Technical form of the active ingredient tested. (for reregistration only)
2. At least 5 young adult rats/sez’groi.ip
3. Dosing, single oral may be administered over 24 his.
4 Vehicle control if other than water.
5. Doses tested, sufficient to determine a toxicity category or a limit dose (5000 mg/kg)
6. Individual observations at least once a day.
7. Observation period to last at least 14 days, or until all test animals appear normal whichever
is longer.
8. — Individual daily observations.
9.’ Individual body weights.
10.’ Gross necropsy on all animals.
Criteria marked with a ‘are supplemental and may not be required for eveiy study.
C-84
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Subdivision F
Guideline Ref. No. 81.1
December 24, 1989
81-1 Acute Oiul Toxicity in the Rat
GUIDANCE FOR SUMMARIZING STUDIES
Items in snrnm ry should indude the items discuased in Qtaptcr 2 of this package and the specific items
listed below.
1. The form of pesticide tested, e.g. solid, liquid, percent Al in technical, etc.
2. The number of animals/dose/sex tested.
3. Dosing route and regimen.
4 Vehicle used
5. Doses tested and results
6. Individual observations on day of dosing.
7. Individual observations on day of dosing and for at least 14 days or until all animals appear normal
(whichever is longer).
8. See items 6 and 7
9. Summarization of body weights
10. Summarization of gross necropsy
11. Significance of changes from the Acceptance Cntena
C-85
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Subdivision F
Guideline Ref. No. 81.2
December 24, 1989
81-2 Acute Dernial Tondty In the Rat, Rabbit or Gmne Pig
ACC FFANCE CRrIERJA
Do vur study meet the tbllowing a ptanee aiteria?
1. Techiucal form of the active ingredient tested. (for reregistration only)
2._ At least 5 animals/sex/group
3.’ . . . . . . .._ Rats 200-300 gm, rabbits 2.0-3.0 kg or guinea pigs 350-450 gin.
4. — Dosing, single dermal.
5. Dosing duration at least 24 hours.
6.* Vehicle control, only if toxicity of vehicle is unknown.
7. Doses tested, sufficient to determine a toxicity category or a limit dose (2000 mg/kg).
8. — Application site clipped or shaved at least 24 hours before dosing
9. — Application site at least 10% of body surface area.
10. — Application site covered with a porous nonimiating cover to retain test material and to
prevent ingestion.
11. Individual observations at least once a day
12. Observation period to last at least 14 days, or until all test animals appear normal whichever
is longer.
13. Individual daily observations.
14.*__ Individual body weights.
15. Gross necropsy on all animals.
Cntena marked with a * are supplemental and may not be required for every study.
C-86
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Subdivision F
Guideline Ret. No. 81-2
December 24, 1989
81-2 A zte Dermal Tonaty in the Rat, Rabbit or Gnine i Pig
GUIDANCE FOR SUMMARJzfl G S’IlJDIES
Items in sninm ry should indude the items d ci sed in Qiapter 2 of th peckage and the specific items
listed below.
1. The form of pesticide tested, e.g., solid, liquid, percent Al in technical, etc.
2. The number of anixnals/sexjdose
3. Weight range of animals
4. Verification of single, dermal exposure
5. Duration of dermal exposure
6. Statement of vehicle control
7. Doses tested and results
8. Preparation of application site
9. Area of application site (percent body surface)
10 Occlusion of test material on application site
11. Individual observations on day of dosing
12. Individual observations on day of dosing and for at least 14 days or until all animals appear normal
(whichever is longer)
13. See items 11 and 12
14 Summanzation of body weights
15. Summarization of gross necropsy
16. Significance of changes from Acceptance Criteria
C -87
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Subdivision F
Guideline Ret. No. 81-3
December 24, 1989
81-3 Acute tnh ilsitiou T uty in the Rat
ACCEPTANCE CR FER1A
Do vur study meet the following a pIanee 1ter a?
1. — Technical form of the active ingredient tested. (for reregistration only)
2. Product is a gas, a solid which may produce a significant vapor hazard based on toxicity and
expected use or contains particles of inhalable size for man (aerodynamic diameter 15 urn or
less).
3• At least 5 young adult rats/sex/group
4._ Dosing, at least 4 hours by inhalation.
5._ Chamber air flow dynamic, at least 10 air changes/hour, at least 19% oxygen content.
6. — Chamber temperature, 22° C a2°), relative humidity 40-60%.
7. Monitor rate of air flow
8. Monitor actual concentrations of test material in breathing zone.
9. Monitor aerodynamic particle size for aerosols.
10. — Doses tested, sufficient to determine a toxicity calagory or a limit dose (5 mgfL actual
concentration of respirable substance).
11. Individual observations at least once a day.
12. Observation penod to last at least 14 da , or until all test animals appear normal whichever
is longer.
13. — Individual daily observations.
14. Individual body weights.
15._ Gross necropsy on all animals.
Criteria marked with a are supplemental and may not be required for every study.
C-88
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Subdivision F
Guideline Ref. No. 81-3
December 24, 1989
81-3 Acute Inhabtion Toxicity in the Rat
GUIDANCE FOR SUMMARI7JNG STUDIES
Items in summziry should indude the items discussed in Chapter 2 of this package and the specific items
listed below.
1. The form of pesticide tested, e.g., solid, liquid, percent Al in technical, etc.
2. Statement of the inhalability of test substance
3. The number of animals/sexjdose
4 Duration of inhalation exposure
5. Number of chamber air changes,ltour and the percent oxygen content of chamber air
6. Ranges for chamber air temperature and relative humidity
7. Air flow rate
8. Analytical concentrations of test material in breathing zone
9. Results of aerosol particle-size determination
10. Doses tested (or limit dose of 5mgfL or highest attainable)
11. Individual observations on day of dosing
12. Individual observations on day of dosing and for at least 14 days or until all animals appear normal
(whichever is longer)
13. See items 11 and 12
14. Summarization of body weights
15. Summarization of gross necropsy
16. Significance of changes from Acceptance Criteria
C-89
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Subdivision F
Guideline Ref No. 81-4
December 24, 1989
81-4 Primaiy Eye hritatlon in the Rabbit
ACCEPTANCE CRITERIA
Does pour study meet the following a p1an aiteiia?
1. — Technical form of the active ingredient tested. (for reregistration only)
2. — Study not required if material is corrosive, causes severe dermal irritation or has a pH of .
2 or . 11.5.
3._ 6 adult rabbits
4. — Dosing, instillation into the conjunctival sac of one eye per animal.
5. _ Dose, 0.1 ml if a liquid; 0.1 ml or not more than 100 trig if a solid, paste or particulate
substance.
6. — Solid or granular test material ground to a fine dust.
7. — Eyes riot washed for at least 24 hours.
8. — Eyes examined and graded for imtation before dosing and at 1, 24, 48 and 72 hr, then daily
until eyes are normal or 21 days (whichever is shorter).
9.’_ Individual observations for the entire day of dosing,
10._ Individual daily observations.
Criteria marked with a are supplemental and may not be required for every study.
C.90
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Subdivision F
Guideline Ref. No. 81.4
December 24, 1989
81-4 Primaiy E Imtauon in the Rabbit
GUIDANCE FOR SUMMARIZJNG STUDIES
Items in snmm ry should indiule the items disc sed in C iapte 2 of th package and the specific items
listed below.
1. The form of pesticide tested, e.g., solid, liquid, percent Al in technical, etc.
2. State of material is corrosive, cause severe dermal imtauon or has a pH of <2 or >11.5
3. Number of adult rabbits tested
4. State method of dosing, i.e., instillation into the conjunctival sac of one eye per animal
5. Dose administered
6. Note whether solid or granular test matenal has been ground to a fine dust
7. State whether eyes were washed and at what time post instillation (not less than 24 hours)
8. State whether eyes were examined and graded for untation before dosing and at what periods after
dosing
9. Individual observations for entire day of dosing
10. Individual observations for entire day of dosing and individual daily observations afterwards, until
eyes are normal or for 21 days
11. Significance of changes from Acceptance Criteria
C-91
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Subdivision F
Guideline Ref. No. 81-5
December 24, 1989
81-5 Primaiy Dermal Irritation Study
ACCEPTANCE CRrIERIA
Does your study m t the following a ptanoe aiteria?
1. — Technical form of the active ingredient tested. (for reregistration only)
2. — Study not required if matenal is corrosive or has a pH of 2 or 11.5.
3* 6 adult animals.
4. Dosing, single dermal.
5. Dosing duration 4 hours.
6. — Application site shaved or clipped at least 24 hour pnor to dosing.
7. — Application site approximately 6 cm 2 .
8. — Application site covered with a gauze patch held in place with nonirritating tape
9. — Material removed, washed with water, without trauma to application site
10 Application site examined and graded for irntation at 1, 24, 48 and 72 hr. then daily until
normal or 14 da (whichever is shorter).
11._ Individual observations for the entire day of dosing.
12.*_ Individual daily observations.
Criteria marked with a * are supplemental and may not be required for evety study.
C-92
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Subdivision F
Guideline Ref. No. 81-5
December 24, 1989
81-5 Primary Dermal lmtation Study
GUIDANCE FOR SUMMARIZm,G STUDIES
Items in summary should indude the items d ed in Chapter 2 of this p ’ttage and the specific items
listed below.
1. The form of pesticide tested, e.g., solid, liquid, percent Al in technical, etc.
2. State if material is corrosive, has a pH <2 or >11.5, or has a dermal LD-50 <200 mg,lcg
3. Number of adult animals tested
4. Amount applied
5 Duration of dermal exposure
6 Preparation of application site (shaved or clipped at specified time before dosing)
7. Area of application site
8. Method for occlusion of application site
9. Note removal of test matenal and if skin was washed with water
10. State times post application when site was graded for irritation
11. Individual observations for entire day of dosing.
12. Individual observations for entire day of dosing and individual daily observations thereafter
13. Significance of changes from Acceptance Criteria.
C-93
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Subdwjs F
Guideline Ref. No. 81-6
December 24, 1989
81-6 Dernial Seti in the Guinea Pig
AC( r cj CRITERIA
Does ur study meet the following a p*anee aiteria?
1. Technical form of the active ingredient tested. (for reregistration only)
2. Study not required if material is corrosive or has a pH of 2 or 11.5.
3. One of the following methods is utilized;
Freund’s complete adjuvant test
— Guinea pig maximization test
Split adjuvant technique
Buehler test
Open epicutaneous test
Mauer optimization test
Footpad technique in guinea pig
Other test accepted by OECD (spec1 )_
4. Complete description of test
5* Reference for test.
6. Test followed essentially as described in reference document.
7.’ Positive control included.
Criteria marked with a * are supplemental and may not be required for every study.
C-94
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Subdivision F
Guideline Ref. No. 81.6
December 24, 1989
814 Dermal Seusitization in the Guinea Pig
GUIDANCE FOR SUMMARIZING S’TUDIES
hems in summ iy should indude the items discussed in ( apter 2 of this package and the specific items
listed bek w.
1. The form of pesticide tested, e.g., solid, liquid, percent A! in technical, etc.
2. State if material is corrosive or has p1-I <2 or >11.5).
3. State specific method utilized
4. Complete description of specific method
5. Reference for the specific method employed
6. Note adherence of the protocol to that in the reference for the specific method utilized
7. State the positive control tested
8. Significance of changes from Acceptance Criteria
C-95
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Subdivision F
Guideline Ref. No. 81-7
December 24, 1989
81-7 Acute Neumtoxidty in the Hen
ACCEPTANCE CRiTERIA
Does your study meet the following a ptanee aiteria?
1. — Study performed on an organophosphate cholinesterase inhibiting compound.
2. — Technical form of the active ingredient tested.
3.’_ Positive control utilized.
4. — Species utilized, domestic laying hen 8-14 months of age.
5. — Dosing oral by gavage or capsule (dernial or inhalation may be used).
6. — An acute oral LD is determined.
7. Dose tested equal to an acute oral LD or a limit test of 5000 mg/kg.
8.’__ Dosed animals may be protected with atropine and/or 2-PAM.
9. Sufficient test animals so that at least 6 survive
10. — Negative (vehicle) control group of at least 6 hens
11. Positive control of at least 4 hens. (if used)
12. — Test dose repeated if no signs of delayed neurotoxicity observed by 21 days after dosing.
13. Observation period 21 days after each dose.
14. Individual daily observations.
15. Individual body weights.
16. Individual necropsy not required.
17. Histopatholo performed on all animals. Tissue to be fixed in situ preferably using whole
animal perfusion techniques. At least three sections of each of the following tissues:
__ . .brain, including medulla oblongata
_spinal cord; upper cervical, mid-thoracic and lurnbro-sacral regions
_tibial nerve; proximal regions and branches
sciauc nerve
Criteria marked with a * are supplemental and may not be required for every study.
C-96
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Subdivision F
Guideline Ref. No. 81.7
December 24, 1989
81-7 Acute Neurotoxicity in the Hen
GUIDANCE FOR SUMMARIZING STUDIES
Items in sumn ry should include the items dis sed in Chapter 2 of this package and the specific items
Listed bek w.
1. Note that the test material is a cholinesterase-inhibiting organophosphate
2. The form of pesticide tested, e.g., solid, liquid, percent Al in technical, etc.
3. The positive control utilized
4 Species utilized (domestic laying hen, 8-14 months of age).
5. Route of exposure
6. Results of acute oral LD-50 test
7. State dose tested (equal to either acute oral LD-50 or limit test of 5,000 mg/kg)
8. Note whether animals are protected with atropine andlor 2-PAM
9 Number of test animals
10 Size of negative (vehicle) control group
11. State use of (optional) positive control
12. Note fulfillment of requirement for repeat dosing if no signs of delayed neurotoxicity by 21 days
13. Observation period (21 days after each dose)
14. Individual daily observations
15 Summary of body weights
16. Individual necropsy not required
17. Histopathology performed on all animals. State tissues examined and number of sections of each
Tissues to be fixed in situ preferably using whole animal perfusion techniques.
18 Significance of changes from Acceptance Criteria
C-97
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Subdivision F
Guideline Ref. No. 82-1
December 24, 1989
82-1 Subcbronic Feeding in the Rodent and Noniodent
ACw’TANCE CRITERIA
Does vur study meet the following a pta a iteña?
1. — Technical form of the active ingredient tested.
2. At least 10 rodents or 4 nonrodents, ex/group (3 test groups and control group).
3. Dosing duration daily for 90-days or 5 daysftveek for 13 weeks.
4. — Doses tested include signs of toxicity at high dose but no lethality in nonrodents or a limit
dose if nontoxic (1000 mg/kg).
5. — Doses tested include a NOEL
6.’_ Analysis for test material stability, homogeneity and concentration in dosing medium
7. — Individual daily observations.
8. Individual body weights.
9. Individual or cage food consumption.
l0.._ Opthalmoscopic examination (at least pretest and at term) control and high dose.
11. — Clinical pathology data of 12 & 13 at termination for rodents; for nonrodents at the
beginning then either monthly or midway and at termination.
12. — Hematology.
Erythrocyte count Leucocyte count
— Hemoglobin _ Differential count
E-Iematocnt — Platelet count (or cloning measure)
13. — Clinical chemistry.
‘ Alkaline phosphatase — Total Protein
— Aspartate aminotransferase — Albumin
_ Creatinine kinase Urea nitrogen
Alanine aminotransferase — Inorganic phosphate
‘ Lactic dehydrogenase Calcium
— Glucose * Potassium
— Bilirubin Sodium
* Cholesterol * Chloride
* Creatinine
14. _ Urinalysis, only when indicated by expected or observed activity. As scheduled in 11.
Blood Total bilirubin
— Protein ‘ Urobilirubin
Ketone bodies — Sediment
— Appearance Specilic gravity (osmolahty)
— Glucose ‘ Volume
15. — individual neczopsy of all animals.
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-98
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Subdivision F
Guideline Ref. No. 82-1
December 24, 1989
— aorta
—eyes
— caecum
— colon
— duodenum
— braint
— heart
— testest
— pituitary
— ileum
— trachea
— gall bladder
— jejunum
bone marrow
liverf
— lung
lymph nodes
stomach
mammary gland
— spleen
musculature
— epididymis
adrenals
unnarv bladder
— accessory sex organs, uterus
f organs to be weighed
Critena marked with a * are supplemental and may not be required for every study.
16. Histopatholo of the following tissues performed on all nonrodents and rodents, all control
and high dose animals, all animals that died or were killed on study, all gross lesions on all
animals, target organs on all animals and lungs, liver and kidneys on all other animals.
peripheral nerve
— kidneyst
— esophagus
ovaries
oviduct
pancreas
rectum
spinal cord (3x)
— thyroid / parathyroids
— salivary glands
Ihymus
c.99
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Subdivision F
Guideline Ref No. 82-1
December 24, 1989
-1 Subchronic Feeding in the Rodent and Nonrodent
GUIDANCE FOR SUMMARizm G STUDIES
Items in Sulninaly should indude the items discussed in aiapter 2 of this package and the specific items
Listed below.
1. The form of the pesticide tested
2. The number of animaIs/dose, tested
3. Dosing duration and days/week
4. Dose tested and results
5. NOELs for various parameters
6. Analysis of test material, stability, homogeneity and concentration in dosing medium
7. Individual daily observations
8. Individual body weights
9. Food consumption
10. Ophthalmoscopic exam results
11. Clinical pathology at various times tested (see items 12 and 13)
12. Hematology results at various times tested
13. Clinical chemistry results at vanous times tested
14. Urinalysis results at various times tested
15. Necropsy results
16. Histopathology (all nonrodents and rodents, all control and high dose animals, all animals that died
or were killed on study, all gross lesions on all animals, target organs on all animals and lungs, liver
and kidneys on all other animals. Provide organ weights.)
17. Significance of changes from the Acceptance Cnteria
C-lao
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Subdivision p
Guideline Ref. No. 82-2
December 24, 1989
f2-2 Repeated l c Dermal Toxiaty (21-day) in the Rat, Rabbit or Guinea Pig
ACCEPTANCE CRITERIA
Does your study meet the following a ptance aiteria?
I. Technical form of the active ingredient tested.
2. — At least 5 animals/sez’group (3 test groups and control group).
3. — Dosing duration at least 6 hour/day for 21 days or 5 days/week for 3 weeks.
4 Application site at Least 10% of body surface area.
5. Doses tested include signs of toxicity at high dose, no or minimal dermal irritation, minimal
lethality or a limit dose (1000mg/kg) if nontoxic.
6’ Doses tested include a NOEL
7. — Individual daily observations.
8. IndividuaL body weights.
9. — Individual or cage food consumption.
10. Clinical pathology data of 11 & 12 at termination.
11. Hematology.
— Eiythrocyte count — Leucocyte count
— Hemoglobin ‘ Differential count
— Hematocnt Platelet count (or clotting measure)
12. — Clinical chemistry.
‘ Alkaline phosphatase Total Protein
— Aspartate aminotransferase — Albumin
— Alanine aminotransferase
‘ Creatinine kinase — Urea nitrogen
‘ Lactic dehydrogenase — Inorganic phosphate
Glucose Calcium
— Bilirubin • Potassium
* Cholesterol — Sodium
‘ Creatinine * Chloride
13.’_ Unnalysis, only when indicated by expected or observed activity. As scheduled in 10.
— Blood — Total bilirubin
— Protein * Urobilirubin
— Ketone bodies Sediment
— Appearance — Specific gravity (osmolality)
— Glwx se ‘ Volume
14. Individual neeropsy of all animals.
15. — Histopathology performed on all control and high dose animals, all animals that died or were
killed on study consisting of all gross lesions on all animals, target organs on all animals (to
determine a NOEL), and skin (normal and treated) lungs, liver and kidneys.
Criteria marked with a • are supplemental and may not be required for every study.
C..101
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Subdwisiofl F
Guideline Ref. No. 82.2
December 24, 1989
gZ-2 Repeated D e Dermal T ty (21-day) in the Rat, Rabbit or Guinea Pig
GUIDANCE FOR SUMMARIZING S11JD ES
Items in snmmaly should include the iteun discussed in Chapter 2 of this pac ge and the specific items
listed below.
1. Form of pesticide tested
2. Number of animals/dose/Sex
3. Duration of dosings (hours/day and days/week)
4. Area of application site (absolute and percent of body surface area)
5. Doses tested and results
6. NOELs for various parameters
7. Daily observations
8. Body weights
9. Food consumption
10. Clinical patholo ’ at various times tested (see items Il and 12)
11. UematoLo results at various times tested
12. Clinical chemistry results at various times tested
13. Urinalysis results at various time tested
14. Necropsy results
15. Histopatholo ’ (all control and high dose animals all animals that died or were killed on study
consisting of all gross lesions on all animals, target organs on all animals and skin [ normal and
ireatedj lungs, liver and kidneys)
16, Significance of changes from the Acceptance Criteria.
C- 102
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Subdivision F
Guideline Ref. No. 82.3
December 24, 1989
82.3 Repeated Dose Dexmal Tonaty ( 9 0-day) in the Rat, Rabbit or Guinea Pig
ACCEPTANCE CRiTERIA
Does ur study m t the following a ptance aiteria?
1. — Technical form of the active ingredient tested.
2. — At least 10 animals/sex/group ( 3 test groups and control group).
3. — Dosing duration at least 6 hour/day daily for 90 days or 5 days/week for 13 weeks.
4. Application site at least 10% of body surface area.
5 — Doses tested include signs of toxicity at high dose, no or minimal dermal imtation, minimal
lethality or a limn dose (1000mg/kg) if nontoxic.
6.’ Doses tested include a NOEL
7. — Individual daily observations.
8. — Individual body weights.
9. — Individual or cage food consumption.
10.’_ Opihalmoscopic examination (at least pretest and at term) control and high dose.
11. — Clinical pathology data of 12 & 13 in all animals at termination.
12. Hematology.
— Erythrocyte count — Leucocyte count
— Hemoglobin ‘_ Differential count
— Hematocnt — Plaielet count (or clotting measure)
13. — Clinical chemistry.
* Alkaline phosphatase Total Protein
— Aspartate aminotransferase — Albumin
— Alanine aminotransferase
Creatinine kinase — Urea nitrogen
‘__ Lactic dehydrogenase — Inorganic phosphate
— Glucose Calcium
— Bilirubin -. Potassium
* Cholesterol — Sodium
‘ Creatinine ‘ Chloride
14. ’_ Urinalysis, only when indicated by expected or observed activity. As scheduled in 11.
Blood — Total bilirubin
— Protein Urobilirubin
— Ketone bodies Sediment
— Appearance — Specific gravity (osmolality)
— Glucose Volume
15. — Individual necropsy of all animals.
Critena marked with a ‘ are supplemental and may not be required for every study.
C-103
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Subdivision F
Guideline Ref. No. 82-3
December 24, 1989
16. Histopathology of the following tissues performed on all nonrodents and rodents, all control
and high dose animals, all animgls that died or were killed on study, all gross lesions on all
animals, target organs on all animals and lungs, liver and kidneys on all other animals.
— aorta — peripheral nerve
— kidneysf
— esophagus
ovaries
oviduct
— pancreas
rectum
spinal cord (3x)
— thyroid / parathyroicls
salivary glands
— ihymus
eyes
— caccum
— colon
— duodenum
— braint
-— skin
— heart
— testesf
— pituitary
—. ileum
— trachea
— gall bladder
t organs to be weighed
Criteria marked with a • are supplemental and may not be required for every study.
— jejunum
bone marrow
— liverf
— lung
— lymph nodes
stomach
— mammary gland
— spleen
musculature
— epididymis
adrenals
unnaiy bladder
accessory organs; uterus
C-104
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Subdivision F
Guideline Ref. No. 82-3
December 24, 1989
-3 Repeated se Dermal T iaty (90-day) in Lhe Rat, Rabbit or Guinea Pig
GUIDANCE FOR SUMMARIZING SI1JDJES
Items in snrnrn y should indude the items dis sed in Chapter 2 of this package and the specific items
Listed below.
1. Form of pesticide tested
2. Number of animals/dose/sex
3. Duration of dosing (hours/day and days/week)
4. Area of application site (absolute and percent of body surface area)
5. Doses tested and results
6. NOELS for various parameters
7. Daily observations
8. Body weights
9. Food consumption
10. Ophihalmoscopic exam
11. Clinical pathology at vanous times tested (see items 12 and 13)
12. Hematology results at vanous times tested
13. Clinical chemistiy results at various times tested
14. Urinalysis results at various times tested
15. Necropsy result
16. Histopathology (all control and high dose animals, all animals that died or were killed on study
consisting of all gross lesions on all animals, target organs on all animals and skin [ normal and
treatedj lungs, liver and kidneys)
17 Significant changes from the Acceptance Critena.
C-105
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Subdivision F
Guideline Ref. No. 82-4
December 24, 1989
S2-4 Subchronic Inhi lation Toxicity (90-day) in the Rat
ACCEPTM4CE CRiTERIA
Does ur study meet the foliowing a ptan aiteria?
1. — Technical form of the active ingredient tested. (for reregistration only)
2. — Product is a gas, a solid which may produce a significant vapor hazard based on toxicity and
expected use or contains particles of inhalable size for man (aerodynamic diameter 15 urn or
less).
3. At least 10 young adult rats/sex/group
4. — Dosing, 6 hours per day, 5 days per week for 13 weeks.
S. Food and water should be withheld during dosing.
6. Chamber air flow dynamic, at least 10 air changesThour, at least 19% oxygen content.
7 — Chamber temperature, 22° C (±2°), relative humidity 40-60%.
8. _ Alternatively, oro-nasal or head only exposures may be used.
9. — Monitor rate of air flow,
10 Monitor actual concentrations of test material in breathing zone.
11. — Monitor aerodynamic particle size for aerosols.
12. — Individual daily observations.
13. — Individual body weights.
14. Individual or cage food consumption.
15. ••••_ Opihalmoscopic examination (at least pretest and at term) control and high dose.
16. Clinical pathology data of 17 & 18 in all animals at termination.
17 — Hematology.
— Erythrocyte count Leucocvte count
Hemoglobin ‘ Differential count
l-’ matocnt Platelet count (or clotting measure)
18. Clinical chemisuy.
Alkaline phosphatase Total Protein
Aspartate aminotransferase Albumin
— Alanine aminotransferase Urea nitrogen
• Creatinine kinase Inorganic phosphate
‘ Lactic dehydrogenase Calcium
Glucose ‘ Potassium
Bilirubin Sodium
‘ Cholesterol • Chloride
* Creatinine
19._ Urinalysis, only when indicated by expected or observed activity. As scheduled in 16.
— Blood — Total bilirubin
Protein s Urobilirubin
Ketone bodies — Sediment
— Appearance Specific gravity (osmolality)
Glucose Volume
20 Individual necropsy of all animals.
Criteria marked with a * are supplemental and may not be required for every study.
C-106
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Subdivision F
Guideline Ref. No 82-4
December 24, 1989
aorta
_eyes
caecum
colon
duodenum
— braint
skin
heart
— testesf
— pituitary
ileum
trachea
— gall bladder
jejunum
bone marrow
— livert
— lung
— lymph nodes
stomach
— mammary gland
— spleen
musculature
— epididymis
ad rena Is
— urinary bladder
f organs to be weighed
Criteria marked with a S are supplemental and may not be required for every study.
21. — Histopatholo of the following tissues performed on all nonrodents and rodents, all control
and high dose animals, all animals that died or were killed on study, all gross lesions on all
animals, target organs on all animals and lungs, liver and kidneys on all other animals.
peripheral nerve
— kidneys
esophagus
ovaries
oviduct
— pancreas
rectum
— spinal cord (3x)
— thyroid / parathyroids
salivary glands
thymus
— accessory sex organs; uterus
C-107
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SUbdivi i F
Guidelme Ref. No. 82-4
December 24. 1989
4 Subdironj tnha tjon Toxicity ( 9 0 .day) in the Rat
GUIDANCE FOR SUMM J j j JDIES
Items in sumlnaiy should indude the iten disa tsed in ( apter 2 of this package and the specific items
listed below.
1. Form of pesticide tested
2. Description of physical state of pesticide
3. Number of animals/d /
4. Duratior of dosing (hours/day, days/week)
5. Access to food and water during dosing
6. Chamber air flow
7. Chamber temperature and relative humidity
8. Alternative types of inhalation exposure, it used, e.g. oro-nasal or head only
9. Rate of air flow
10. Actual concent arions of test material in breathing zone
11. Aerodynamic particle size (for aerosols)
12, Daily observations
13. Body weights
14. Food consumption
15. Ophthalmoscopic exams
16. Clinical pathology at various times tested (see items 17 and 18)
17. Hematology results at various times tested
18 Clinical chemistry results at various times tested
19. UrinaJy s results at various times tested
20. Necropsy results
21. Histopathology (all nonrodents and rodents, all control and high dose animals, all animals that died
or were killed on study, all gross lesions on all animals, target organs on all animals and lungs, liver
and kidneys on all other animals. Provide organ weights.
22. Significance of changes from the Acceptance Criteria.
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Subdivision F
Guideline Ref. No. 82.5
December 24, 1989
82-5 Subdironic Neurotozidty (90-day) in the Hen
ACCEPTANCE CRITERIA
Does >vur study meet the following a p1anee aiteria?
1. — Study performed on an organophosphate cholinesterase inhibiting compound.
2._ Technical form of the active ingredient tested.
3. — Positive control utilized. (recommended but optional)
4. — Species utilized, domestic laying hen 8-14 months of age.
5. — At least 10 animals/sex/group [ 3 test groups, a positive control (optional) and a negative
(vehicle) control group].
6. — Dosing duration at least daily for 90 days or 5 days/week for 13 weeks.
7. — Dose route oral gavage or capsule. (dermal or inhalation may be appropriate)
8. Doses tested include signs of toxicity at high dose, no or minimal lethality
9.. Doses tested include a NOEL
10. — Individual daily observations.
11. — Individual body weights.
12. — Individual or cage food consumption.
13. Individual necropsy not required.
14. — Histopatholo performed on all animals. Tissue to be fixed in situ preferably using whole
animal perfusion techniques. At least three sections of each of the following tissues:
— brain, including medulla oblongata
— spinal cord; upper cervical, mid-thoracic and lumbro-sacral regions
— tibial nerve; proximal regions and branches
sciatic nerve
Criteria marked with a are supplemental and may not be required for every study.
C -109
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Subdivision F
Guideline Ref. No. 82.5
December 24, 1989
82-5 Subdironic Neurou dty (90-day) in the Hen
G1J1DANC FOR SUMMARIZING SrUDIES
Items in snmm iy should include the items d cussed in (laptcr2 of ih pacbge and the specific items
listed below.
1. Type of chemical structure
2. Form of pesticide tested
3. Positive control (if used)
4. Species and age
5. Number of animals/dose/sex tested
6. Dosing duration
7. Dosing route
8. Doses tested and results
9. NOELs for various parameter
10. Daily observations
11. Body weights
12. Food consumption
13. Necropsy (if done)
14. Histopatholo i (all animals providing a full description of perfusion and fixation techniques (or the
key tissues of the nervous system)
15. Significant changes from the Acceptance Criteria.
c-Ito
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Subdivision F
Guideline Ref. No. 83-1
December 24, 1989
83-i aironic Feeding in the Rodent and Nonrodent
ACCEPTANCE CRITERIA
Does your study meet the following a psance afteria?
1. — Technical form of the active ingredient tested.
2. — At least 20 rodents or 4 nonrodents/sex/group (3 test groups and control group).
3. — Dosing duration in rodents minimum 12 month nonfood use, 24 months food use; in
nonrodents minimum 12 months 1 .
4. — Doses tested include signs of toxicity at high dose but no lethality in nonrodents or a limit
dose if nontoxic (1,000 mg/kg).
5.’ Doses tested include a NOEL
6 * Analysis for test material stability, homogeneity and concentration in dosing medium
7. — Individual daily observations.
8. — Individual body weights.
9. Individual or cage food consumption.
1O. ’••••_ Opthalmoscopic examination (at least pertest and at term) control and high dose.
11. — Clinical patholo ’ data for all nonrodents and at least 10 rodents/group consisting of 12, 13
&14.
12. — Hematology at 6 month intervals consisting of at least;
— Erythrocyte count — Leucocyte count
— Hemoglobin Differential count
Hematocrit — Platelet count (or clotting measure)
13. — Clinical chemistry at 6 month intervals consisting of at least;
_ Alkaline phosphatase — Total Protein
Aspartate aminotransferase Albumin
— Alanine aminotransferase — Urea nitrogen
‘ Creatinine kinase — Inorganic phosphate
‘_ Lactic dehydrogenase — Calcium
— Glucose ‘ Potassium
— Bi lirubin — Sodium
Cholesterol ‘ Chloride
* Creatinine
14. — Urinalysis at 6 month intervals consisting of at least;
— Blood Total bilirubin
Protein ‘ Urobilirubjn
Ketone bodies - — Sediment
Appearance — Specific gravity (osmolality)
— Glwxise ‘ Volume
15. — Individual necropsy of all animals.
Criteria marked wuh a are supplemental and may not be required for every study.
C-ill
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Subdivision F
Guideline Ref. No. 83-i
December 24, 1989
16. Histopathology of the following tissues performed on all nonrodents and rodents, all control
and high dose animals, all animals that died or were killed on study, all gross lesions on all
animals, target organs on all animals and lungs, liver and kidneys on all other animals.
peripheral nerve
kidneysf
esophagus
ova ries
oviduct
pancreas
rectum
— spinal cord (3x)
thyroid I parathyroids
— salivary glands
— thymus
accessory sex organs; uterus
aorta
_eyes
— caecum
— colon
— duodenum
— brainf
skin
— heart
testest
— pituitary
— ileum
— trachea
— gall bladder
organs to be weighed
l En some cases, a six month study may be acceptable. Contact EPA to discuss the critena for
determining if such a study can be used to support reregistration.
Criteria marked with a * are supplemental and may not be required for every study.
— jejunum
bone marrow
— liverf
— lung
lymph nodes
stomach
— mammary gland
— spleen
musculature
— epididymis
adrenals
— urinary bladder
C-i 12
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Subdivision F
Guideline Ref. No. 83-1
December 24, 1989
$3-i (]ironjc Feeding in the Rodent and Nonrodent
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the iten d amsed in Chapter 2 of this package and the spei fic items
listed below.
1. Form of pesticide tested.
2. Number of animals/dosei ex tested.
3. Dosing duration.
4. Doses tested and results.
5. NOEL for various parameters.
6. Analysis of test material, stability, homogeneity.
7. Daily observations.
8. Body weights.
9. Food consumption
10. Ophthalmoscopic examination.
11. Clinical patholo ’ including hematolo ’, clinical chemistry and urinalysis.
12. Necropsy results.
13. Histopatholo i (all interim sacrifice, all controls high dose animals, all animals that died or were
killed on study, all gross lesions on all animals, target organs on all animals and lungs, liver and
kidneys on all other animals).
14 Significant changes from the Acceptance Critena.
C-i 13
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Subdivision F
Guideline Ref. No. 83-2
December 24, 1989
&3-2 Ona’genicity in Rats or Miee
ACCEPTANCE CRJThRIA
Does ur study meet the following a ptance criteria?
1. Technical form of the active ingredient tested.
2. At least 50 animals/sexigroup (3 test groups and control group).
3. Dosing duration is at least 18 months for mice and 24 months for rats.
4. Number of survivors in any group does not fall below 50% at 15 months for mice, 18
months for rats or 25% at 18 months for mice, 24 months for rats.
54 Doses tested include an MTD or limit dose if nontoxac (1,000 mg/kg).
6.* Doses tested include a NOEL for systematic effects.
7.” Analysis for test material stability, homogeneity and concentration in dosing medium
8. Individual daily observations.
9. Individual body weights.
10. Individual or cage food consumption.
11. — Individual necropsy of all animals.
12. — Blood smear from 10 aninials/se kjose at 12 and 18 months and termination. Differential
count high dose and controls, all other doses if high dose shows pathology.
13. Histopathology of the following tissues performed on all interim sacrifice animals, all control
and high dose animals, all animals that died or were killed on study, all gross lesions on all
animals, target organs on all animals and lungs, liver and kidneys on all other animals
— aorta — jejunum peripheral nerve
— eyes — bone marrow — kidneysf
— caecum — livert esophagus
— colon — lung ovaries
duodenum — lymph nodes — oviduct
— braint — stomach — pancreas
— skin mammary gland — rectum
— heart spleen — spinal cord (3x)
— testest — musculature thyToid / parathyroids
— pituitary — epididymis — salivary glands
— ileum adrenals — thymus
— trachea urinary bladder accessory sex organs; uterus
gall bladder
t organs to be weighed
The position document entitled “Selection of a Maximum Tolerated Dose (MTD) in
Oncogemcity Studies (EPA No. 540i09-88-003) stated EPA’s criteria for determining if an
oncongenicity study has been adequately performed in terms of doses tested. However OPP
is also aware that older oncogenicity studies, upon initial review or re-review. may have been
tested at doses lower than the predicted MTD. In the event that such testing appears to be
at doses less than the predicted MTD, the Office of Pesticides Program has been reviewing
Criteria marked with a * are supplemental and may not be required for every study.
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Subdivision F
Guideline Ref. No. 83-2
December 24, 1989
demonstrated oncogenicity in another species, nearness to the apparent MTD, genotoxic effects,
structure-activity factors, absolute value of the highest dose tested and metabolic considerations.
Criteria marked with a * are supplemental and may not be required for every study.
c-i 15
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Subdivision F
Guideline Ref. No. 83.2
December 24, 1989
*3.2 On genicaty in Rats or Mux
GUIDANCE FOR SUMMARIZING STUDIES
Items in summaly should indude the itema dsansesl in Chapter 2 of this package and the specific items
listed below.
1. Form of pesticide tested
2. Number of animals/dose/sex
3. Duration of dosing
4. Survival rates
5. Descnption of effects at highest dose tested
6. NOELs for various parameters
7, Analysts of test material, stability, homogeneity, and concentration in dosing medium
8. Daily observations
9. Body weights
10. Food consumption
Ii. Necropsy results
12. Blood smear results
13. 1-listopathology (all interim sacnfice, all controls and high dose animals that died or were killed on
study, all gross lesions on all animals, target organs on all animals, and lungs, liver, kidneys on all
other animals.)
14. Significance of changes from the Acceptance Criteria
C .116
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Subdivision F
Guideline Ref. No. 83-3
December 24, 1989
83-3 Teratolo , Studi
ACCEPTANCE CRIThRIA
Does your study meet the Ibilowing a ptanir aiteria?
1. Technical form of the active ingredient tested.
2. At least 20 pregnant animals/dose group for mice, rats or hamsters are available. At least 12
pregnant animals/dose group for rabbits are available (three test groups and control group).
3. — At the high dose, overt maternal effects such as slight weight loss are reported (or a limit
dose is given, 1,000 mg/kg).
4. At the low dose, no developmental toxicity is reported.
5. — Dosing duration is at least during the period of major organogenesis, but may extend up to
one day prior to term.
6 _ Analysis for test material stability, homogeneity and concentration in dosing medium
7. — Individual daily observations.
8. — Individual body weights.
9. — Individual food consumption.
10. — Necropsy on all animals
11. Individual uterine examination including number of fetal deaths, early and late resorptions
and numbers of viable fetuses per sex.
12. All ovaries examined to determine number of corpora lutea.
13. Individual litter weights and/or individual fetal weights per sex/litter.
14. Individual fetus external examination.
IS — Individual fetus skeletal examination for 1/3 to 1/2 of each litter for rodents and all for all
rabbits.
16. — Individual fetus soft tissue examination.
Criteria marked with a * are supplemental and may not be required for every study.
C..117
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Subdivision F
Guideline Ref. No. 83-3
December 24, 1989
&3-3 Teratology Studies
GUIDANCE FOR SUMMARIZING STUDIES
Items in snmmary should indude the items discussed in Chapter 2 of this package and the specific items
listed below.
1. Form of pesticide tested.
2. Number of animals per dose and number of dose groups.
3. Number of litters per dose group.
4. Toxic signs in the dams of the high dose.
5. NOEL for developmental toxicity.
6. Dosing duration and dates.
7. Test material stability, homogeneity and concentration in the dosing medium.
8. Daily observations.
9. Body weight.
10. Food consumption.
11 Results of uterine examination (for fetal deaths, resorption and viable fetuses).
12. Results of ovarian examination (for corpora lutea).
13 Litter and fetal weight.
14. Results of fetal examination (external, skeletal and soft tissues).
15. Significance of changes form the Acceptance Criteria.
C-hg
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Subdivision F
Guideline Ref. No. 83-4
December 24, 1989
-4 Reproduction
ACCEP’FANCE CRITERIA
Does vur study meet the f llowing a pLanoe aneria?
1. Technical form of the active ingredient tested
2. — At least 20 males and sufficient females to yield 20 pregnant /dose group
3. — At least 3 dose groups and a control.
4. — At the high dose, parental toxicity is observed (or a limit dose is given, 1,000 mg/kg/day).
5.’ At the low dose, no reproductive effects are observed.
6.’ Analysis for test material stability, homogeneity and concentration in dosing medium
7. P 1 animals 8 weeks old at the start of the study
8. Dosing is continuous starting with the P 1 animals until an individual animal is sacnficed.
9. Mating is I male to I female.
10. — The mating period is not more than 3 weeks.
11. At least two generations are bred.
12. — Individual daily observations.
13. Individual body weights.
14. Individual food consumption.
15. Individual litter observations.
16. — individual litter weights (pup weights) at birth and on days 4, 7 (optional), 14 and 21
17. Sacrifice schedule, all mating males immediately after last mating, all breeding females
immediately after weaning last litter, all animals not used for breeding immediately after
weaning.
18 e Necropsy on all animals
19 .•• Histopatholo of reproductive organs from all animals on the high dose and control P 1 and
F 1 animals selected for mating. Animals from all other dosing groups if histological effects
are observed at the high dose.
20.’_ Histopatholo of all organs with gross lesions.
Criteria marked with a ‘ are supplemental and may not be required for every study.
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Subdivision F
Guideline Ref. No. 83.4
December 24, 1989
-4 Reproduction
GUIDANCE FOR SUMMARIZING UDIES
items in summary should indude the Items d cussed in Chapter 2 of this package and the sp ific items
Listed below.
1. Form of pesticide tested.
2. Number of dose groups and number of animals per group.
3. Age of the parent animals.
4. Signs of toxicity in the high dose group.
5. NOEL for reproductive toxicity.
6. Analysis of test material stability, homogeneity and concentration in dosing medium.
7. Duration of dosing.
8. Matrngs (number of females per a male, mating period).
9. Study duration (number of generations monitored).
10. Daily observations.
ii. Body weights.
12. Food consumption.
13. Litter observations.
14 Litter weights.
15. Sacrifice schedule.
16. Necropsy.
11 Histopathology of reproductive system and all organs with gross lesions.
18. Significance of changes from the Acceptance Critena.
C.120
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Subdivision F
Guideline Ref. No. 83-5
December 24, 1989
&3. .5 aironic ng noogenicity in the Rat
ACCEPTANCE CR FERIA
Does your study meet the following a ptance aitena?
1. — Technical form of the active ingredient tested.
2. — At least 50 rats/sex/group (3 test groups and control group).
3. — Dosing duration is at least 24 months.
4. Number of survivors in any group does not fall below 50% at 18 months or 25% at 24
months.
54_ Doses tested include an MTD or limit dose if nontoxic (1000 mg/lcg).
6._ Doses tested include a NOEL
Analysis for test material stability, homogeneity and concentration in dosing medium
8. — Individual daily observations.
9. — Individual body weights.
10. Individual or cage food consumption.
11. ••_ Opihalmoscopic examination (at least pertest and at term) control and high dose
12. — Clinical patholo data for at least 10 rats/group consisting of 13, 14 & 15
13. Hematolo at 6 month intervals consisting of at least;
— Erythrocyie count Leucocyte count
— Hemoglobin _ Differential count
— Hematocnt — Platelet count (or clotting measure)
14. — Clinical chemistiy at 6 month intervals consisting of at least;
.__ Alkaline phosphatase Total Protein
Aspartate aminotransferase Albumin
— Alanine aminotransferase — Urea nitrogen
‘ •_ Creatinine kinase Inorganic phosphate
Lactic dehydrogenase — Calcium
— Glucose Potassium
— Bilirubin — Sodium
Cholesterol ‘_ Chloride
* Creatinine
15 — Unnalysts at 6 month intervals consisting of at least;
— Blood — Total bilirubin
— Protein Urobilirubin
— Ketone bodies — Sediment
— Appearance — Specific gravity (osmolality)
Glucose _ Volume
16. Individual necropsy of all animals.
Criteria marked with a are supplemental and may not be required for evety study.
C. 121
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— aorta
_eyes
— caecum
— colon
— duodenum
— brainf
_skin
— hean
— testesf
— pituitary
— ileum
— trachea
— gall bladder
— jejunum
bone marrow
— livert
— lung
— lymph nodes
stomach
— mammary gland
— spleen
musculature
— epididymis
adrenals
— urinary bladder
peripheral nerve
— k.idneyst
— esophagus
ovaries
oviduct
— pancreas
rectum
— spinal cord (3x)
— thyroid I parathyroids
— salivary glands
— thymus
— accessory sex organs; uterus
t organs to be weighed.
The position document entitled Selection of a Maximum Tolerated Dose (MTD) in
Oncogenicity Studies (EPA No. 540109-88-003) stated EPA’s criteria for determining if an
oncongenicity study has been adequately performed in terms of doses tested. However OFF
is also aware that older oncogenicity studies, upon initial review or re-review. may have been
tested at doses lower than the predicted MiD. In the event that such testing appears to be
at doses less than the predicted MTD, the Office of Pesticides Program has been reviewing
and considering the entire weight of the evidence to determine if retesting is necessary
Certain factors which affect the agency’s decision to retest include but are not limited to the
following: demonstrated oncogenicity in another species, nearness to the apparent MTD,
genoloxic effects, structure-activity factors, absolute value of the highest dose tested and
metabolic considerations.
Criteria marked with a are supplemental and may not be required for every study.
Subdivision F
Guideline Ref. No. 83-5
December 24, 1989
17 Histopatholo of the following tissues performed on all nonrodents and rodents, all control
and high dose animals, all animals that died or were killed on study, all gross lesions on all
animals, target organs on all animals and lungs, liver and kidne on all other animals.
C-122
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Subdivision F
Guideline Ref. No. 83-5
December 24, 1989
83-5 C ronic F eding,Oncogenicity in the Rat
GUIDANCE FOR SUMMARIZING STUDIES
Items in snmmary should indude the ite d c sed in Chapter 2 of this package and the specific items
listed below.
1. Form of pesticide tested.
2. Number of animals per dose per sex.
3. Duration of dosing.
4. Survival rate.
5. Toxic signs at the high dose.
6 Dosing regimen and NOEL for various parameters.
7. Analysis of test matenal stability, homogeneity and concentration in dosing medium.
8. Daily observations.
9. Body weights.
10. Food consumption.
11. Ophthalmoscopic examination.
12. Clinical pathology (including hematology, clinical chemistry, and urinalysis).
13. Necropsy results.
14. Histopathology (all interim sacrifice, all controls and high dose animals that dies or were killed on
study, all gross lesions on all animals, target organs on all animals and lungs, liver and kidneys on
all other animals).
15 Significance of changes from the Acceptance Criteria.
C- 123
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Subdivision F
Guideline Ref. No. 84.2
December 24, 1989
84-2 Mutagenidty Studies
ACCEPTANCE CRITERIA
Does your study meet the (oflowing a p ance criteria?
General Requirements
I. Techni l form of the active ingredient teSted.
2. — Negative, solvent and/or vehicle control(s) for the test system.
3. — Positive control(s) for (he test system.
4. — Fully identified test system, species, strain, source etc.
S. Fully described method for maintaining test system.
— Fully described method (or prepanng test environment and adminIstering test coinpond.
7. Fully described metabolic actrvauon system, if required.
8. — Determination of maximum and range of concentrations/doses used under test conditions.
9.’_ Criteria for determination of a positive effect.
Test SDeciflc Requirements
Salmonella reverse mutation assay
1. — Minimum of four strains, TAS8, TAIOO, TA1535 and TA153 . (alternatives need rationale)
2. Strarn specific positive controls.
3. — Highest concentration limited by toxicity, solubitity or 5000 ug/plate.
4. _ At least 5 different conceniranons of test material at adequate inIer rals.
5 ‘ A single positive response confirmed by testing over a narrow range of concentrations.
6.’_ At least three plates experimantal point.
Gene mutation in somatic cells in culture
1. — Highest concentration limited by toxicity (10-20% relative survival), solubility or 5000 ug/mI.
2._ At least 4 different concentrations of test material to yield a concentranon related toxic
effect.
3. — Determination of the number of cell cultures used.
In vitro mammalian cvtogenctics
1. — Highest concentration limited by toxicity (e.g. reduced initotic activity; alteration of cell cycle;
cytotoxicity), solubility or 5000 ug,’tnl.
2.’_ Multiple concentrilons used to define the response.
3 ‘ .... ... At least two independent cultures for each experimental point.
4 Determination of culture harvest time.
Criteria marked with a * are supplemental and may not be required for every study.
C-124
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Subdivision F
Guideline Ref. No. 84-2
December 24, 1989
In vivo mammalian cvtogenetj - bone marrow
1. — At least 5 male and 5 female animals per experimental group.
2. — Highest dose limited by toxicity or 5000 mg/kg.
3. Determination of sampling times.
Aberrations; a) one treatment - 3 times in range of 6-48 hours after treatment adequately
spaced with central sample at 24 hour (may be altered based on cell cycle time). b)
repeated treatments - samples taken 6 and 24 hours after last treatment (may be
altered based on cell cycle time).
Micronucleus; Samples taken 3 times, starting not earlier than 12 hours after the last
treatment and at appropriate intervals following the first sample, but not beyond 72
hours.
4. Micronucleus assay, at least 1000 polychromauc erythrocytes/animal scored. Ratio of poly to
normochromatic determined by counting 200-1000 eiythrocytes (1000 OECD).
Rodent dominant lethal assay
1. Sufficient number of dosed males to provide a minimum of 30 pregnant females per mating
interval.
2. — Concurrent positive control or results from positive control conducted within 12 months in
same laboratoiy with same strain.
3. — Highest dose produced toxicity or 5000 mg/kg.
4. — Sampling or exposure over entire spermatogenesis cycle of dosed males (8 weeks mice, 10
weeks rats)
Mv mutagenicity test with suggestive or greater positive results/activity shall be submitted
reqardless of missing essential items.
Criteria marked with a * are supplemental and may not be required for evety study.
C- 125
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SUIXI IV1SLOa F
Guideline Ref. No. 84-2
December 24, 1989
$4-2 Mutagenicity Studies
GUiDANCE FOR SUMMARIZiNG STUDIES
Items in sn mary should Indude the items d a sed in Chapter2of th pa ge and the specific items
listed below.
General Requirements
1. Form of pesticide tested.
2. Negative, solvent and/or vehicle control(s).
3. Positive control(s).
4. Test system (species, strain, source...etc.)
5. Methods used (for maintaining test system. preparing test environment, administrating test
compound, preparation of metabolic activation system).
6. Range finding.
7. Cntena for determining positive response.
Specific Requirements
Salmonella reverse mutation
1.Strains used.
2.Positive controls.
3.Doses tested and limitations (toxicity, solubility).
4.Number of replicates.
Gene mutation in somatic cells in culture
1 Doses tested and limitations (toxicity, solubility)
2.Number of cell cultures used.
In vitro mammalian cytoaenetics
iDoses tested and hmflations (toxicity, solubility).
2.Number of cultures for each experimental point.
3.Culture harvest.
In viva mammalian cytojenetics - bone marrow
iNumber of animals per sex per dose.
2.Highest dose tested.
3.Sampling time.
Rodent d 9 min . ntjetha1 assay
1.Numb r of pregnant females per mating interval.
2.Concuryent positive control.
3.Doses tested and limitations (Toxicity).
4.Sampllng or esposure over entire spermatogenesis cycle.
C- 126
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Subdivision F
Guideline Ref. No. 85.1
December 24, 1989
85-1 Metabolism Studies
ACCEPTANCE CRiTERIA
Does vur study meet (be ibilowing a ptance aiteria?
1. — Analytically pure grade of the active ingredient.
2. — Isotopically labeled in the core of the molecule and/or significant portions thereof.
-OR-
3. — Analytical procedures sufficiently specific and sensitive to identify the test substance.
4. — Young adult rats. Other mammalian species may be used for specific purposes.
5 — Five male and five female rats for each dose, 4 if following OECD protocol.
6. — Two doses, the low to be without effect and the high to produce toxic or pharmacological
signs but not severe effects or mortality.
7 _ Dosing group A, single low dose by intravenous route (not required if insoluble in water or
normal saline).
8. — Dosing group B, single low dose by oral route.
9 — Dosing group C, 14 consecutive daily low dose of the unlabeled test material by oral route
followed by a single low dose of the labeled test material.
10 — Dosing group D, single high dose by oral route
11. — Collect individually all unne, feces and expired air for 7 days after labeled dose or until 90+
percent of the dose is excreted (whichever occurs first). Expired air not required if a pilot
study shows no excretion in 24 hours.
12. For dosing groups B, C and D. quantity of label in the following tissues and organs.
— bone liver
— brain — lung
— fat — blood
— testes — muscle
— heart spleen
— kidney — residual carcass
— tissues showing pathology in this or pnor studies
For all dosing groups:
13. — Quantities of label in urine, feces and expired air (if detected in preliminary study) at
appropriate intervals (e.g. 4, 8, 12 and 24 hours, 1.5, 2, 3, 4, 5, 6 and 7 days.
14 — Qualitative analysis of urine and feces to detect metabolism and identify metabolites (pooled
urine and feces by dosing group may be used).
NOTE The metabolism data requirement may be filled in part. For example performing the analysis on
a single dose group can satisfy the requirement for that dose.
Criteria marked with a * are supplemental and may not be required for every study.
C-127
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Subdivision F
Guideline Ref. No. 85-1
December 24, 1989
85-1 Metabolj m Studies
GUIDANCE FOR SUMMARIZING STUDIES
heals in snmlnary should include the itenls discussed in aiapter 2 of this package and the specific items
listed below.
1. Form of pesticide used.
2. Radioactive label used and site of labeling.
3. Type and specificity of analytical procedures.
4. Animals species, strain and age.
5. Number of animals per sex per test group.
6. Number of doses tested.
7. Route of administration.
8. Sample collection and schedule.
9. Results of the qualitative and quantitative analysis of urine, feces, tissues, and expired air.
C-128
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SUBDIVISION J
122-1(a) Seed GcrminationIseedling Emergence - Tier I . . . . 130
122-1(b) Vegetative Vigor. Tier I 133
122-2 Growth and Reproduction of Aquatic Plants -. Tier I 136
123-1(a) Seed GerminationiSeedling Emergence -- Tier II . 139
123-1(b) Vegetative Vigor -- Tier II . 142
12.3-2 Growth and Reproduction of Aquatic Plants - . Tier H 145
124-1 Terrestrial Field Test -- Tier HI 148
124-2 Aquatic Field Test -- Tier III 150
C-129
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Subdivision J
Guideline Ref. No. 122-1(a)
December 24, 1989
122-1(a) Seed GerminatioiiiSeeiling Emergence - Tier I
ACCEPTANCE CRIThRIA
Does ur study meet the following ptance cmena?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2. Identification of test substance
— Source, purity, identification number, percent active ingredient
Identity and composition of Impurities
— Water solubility of test material reported in ppm
Vapor pressure of test material
3. Preparation of test solution (control/treatment) described
Solvent used and amount used to dissolve test material
If used, amount of solvent added to control
Water source and description
* Total amount of test material used
‘__ Temperature of water reported
4 Test Plants
Six dictyledoneae species from at least four families and four monocotyledoneae
- species from at least two families are required. Corn, soybean, and a root crop must
be tested
— Family, genus, species, and cultivar or variety
5 Composition and size of test vessels
Test vessels type, size, and shape
Depth of medium in relation to test vessel
6. Test System described
Criteria used to determine effects (i.e. phytotoxicity rating scale)
Critena marked with a * are supplemental and may not be required for every study.
C-130
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Subdivision J
Guideline Ref. No. 122-1(a)
December 24, 1989
7. Test design followed
— Test duration and observation periods live days for seed germination, and two weeks
for seedling emergence
— Dose rate in ppm (seed germination) and pounds of Al per acre (seedling
emergence) at the maximum label rate
— Method of introducing the test solution into the test vessels
— Type and number of controls
— Number of replicates (at least 3)
— Substrate charactensti [ name/description, physical and chemical properties including
pH and % organic matter (substrate should contain no more than 3% organic
matter)J and preparation technique
— Method used to assign test organisms to test and control groups
Number of seeds planted/container (at least 10)
— Date of planting
— Descnption of growth chamber, greenhouse (include size and location), or field plot
— Environmental conditions [ thermoperiod, relative humidity, lighting regime (intensity,
quality,and photoperiod), and watering regimej at the test location
Cultural practices
Method of statistical analysis
Identification of protocol followed (or include copy)
DISCUSSION AND RESULTS
8. — Phytotoxicity symptoms observed
9 — Percent germination and percent emergence at the conclusion of test as compared to controls
10. — Deviations from EPA Subdivision J Guidelines and protocol
11 Conclusions reached based on the results of the study
12. The relationship between the results of the study and the test chemical described, including
Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
Rationale for deviations from the protocol
Criteria marked with a are supplemental and may not be required for every study.
C- 131
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Subdivision J
Guideline Ref. No. 122.1(a)
December 24, 1989
122-1(a) Seed GerminationiSeedling Emergenee - Tier I
GUIDANCE FOR SUMMARIZING STUDIES
Items in sumn ry should Indude the items discmsed in a apter2ot this package and the specific items
Listed bekyw.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of zest species, including
Family, genus, species, and c iltivar or variety names
3. Description of test vessels, including
Test vessel type, size, and shape
4. Description of test system, including
- Criteria used to determine effects (i.e phytotoicicity rating system)
S. Description of test design, including
• Test duration and observation periods
- Dose rate in ppm (seed germination) and pounds of Al per acre (seedling emergence)
Date of planting
DISCUSSION AND RESULTS
6. Phytotoxicity symptoms observed
7. Percent germination and percent emergence at the conclusion of test as compared to controls
8. Conclusions reached based on the results of the study
C-132
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Subdivision 3
Guideline Ref. No. 122.1(b)
December 24, 1989
lfl-1(b) Vegetative Vigor - Tier I
ACCEPrANCE CRITERIA
l es )VUT study meet the foll ing a pIanee aiteria?
GENERAL INFORMATION
1. — Date 01’ test initiation and termination reported
MATERIALS/METHODS
2. Identification of test substance
— Source, purity, identification number, percent active ingredient
Identity and composition of impunties
— Water solubility of test material reported in ppm
— Vapor pressure of test matenal
3. Preparation of test solution (control/treatment) described
— Solvent used and amount used to dissolve test material
If used, amount of solvent added to control
— Water source and descnption
_ Total amount of test material used
Temperature of water reported
4 Test Plants
— Six dicotyledoneae species from at least four families and four monocotyledoneae
species from at least two families are required. Corn, soybean, and a root crop must
be tested.
— Family, genus, species, and cultivar, or variety
Source
5. Composition and size of test vessels
— Test vessel type, size, and shape
— Depth of medium in relation to test vessel
6. Test system described
— Criteria used to determine effects (i.e phytotoxicity rating system)
Cnteria marked with a ‘ are supplemental and may not be required for evety study.
C-133
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Subdivision i
Guideline Ref. No. 122.1(b)
December 24, 1989
7. Test design followed
— Test duration and observation periods (at least 2-weeks with weekly observations)
Dose rate in pounds of Al per acre at the maximum label rate
— Method of foliar application
— Type and number of controls
Number of replicates (at least 3)
— Substrate characterisues [ name/description, physical and chemical properties including
p1-1 and % organic matter (substrate should contain no more than 3% organic
matter)I and preparation technique
— Method used to assign test organisms to test and control groups
Number of plants/container (at least 5)
Date of planting
Description of growth chamber, greenhouse (include size and location) or field plot
Environmental conditions [ thermoperiod, relative humidity, ltghting regime (intensity,
quality, and photopenoct), and watering regimej at the test location
Cultural practices
Method of statistical analysis
Identification of protocol followed (or include copy)
DISCUSSION AND RESULTS
8. — Phytoto icity symptoms observed.
9 — Growth measurements taken (height and dry weight).
10 — Percent effect as compared to control
11. — Deviations from EPA Subdivision J Guidelines and protocol.
12. Conclusions reached based on the results of the study.
13. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
Rationale for deviations from the protocol
Critena marked with a * are supplemental and may not be required for every study.
C.134
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Subdivision J
Guideline Ref. No. 122-1(b)
December 24, 1989
122-1(b) Vegetative Vigor - Tier I
GUIDANCE FOR SUMMARIZING S11JD [ ES
Items in sun’m ry should indude the items disa ed in Chapter 2 of this package and the specific items
Listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test organisms, including
- Family, genus, species, and cultivar or vanety names
3. Descnption of the composition and size of test vessels,including
- Test vessel type, size, and shape
4. Descnption of the test system, including
- Cnteria used to determine effects (i e. phytotoxicitv rating system)
5 Descnption of test design followed
- Test duration and observation periods
- Dose rate in ppm and pounds of Al per acre
- Date of planting
DISCUSSION AND RESULTS
6. Phytotoxicuy symptoms observed
7. Growth measurements taken (height and dry weight)
8. Percent effect compared to control
9. Conclusions reached based on the results of the study
C-135
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Subdivision J
Guideline Ref. No. 122-2
December 24, 1989
122-2 Growth and Reproduction of Aquatic Plants - fler I
ACCEPrANCE CRITERIA
Does )VUZ study meet the following a p1anee aitcria?
GENERAL INFORMATION
1. Data of test initiation and termination reported.
MATERIALS/METHODS
2. Identification of test substance
— Source, purity, identification number, percent active ingredient
Identity and composition of impurities
Water solubility of test material reported in ppm
3. Preparation of test solution (control/treatment) described
Solvent used and amount used to dissolve test material
If used, amount of solvent added to control
Water source and description
Total amount of test material used
Temperature of water reponed
4. Test Plants
Green alga Selenastrum capricornurum , blue-green alga Anabaena flos-aguae ,
duckweed Lemna gibba . marine diatom Skeletonema costatum , and a freshwater
diatom. Include culture type and strain (Type(s) tested dependent on pesticidal
properties and use sites).
History of test organisms described
Source and acclimation described
5. Composition and size of test vessels described
Test vessel type, size, and shape
Ratio of test vessel size to medium
6. Test system described
— Criteria used to determine effects (i.e phytotoxicity rating system)
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-136
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Subdivision .1
Guideline Ref. No. 122-2
December 24, 1989
7. Test design followed
Test duration and observation periods (algae tests at least 5 days with daily
observations, Lemna test at least 14 days with 3 day observations)
— Method of introducing the test solution into the test vessels
— Dose rate in ppm at the maximum label rate
— Type and number of controls
— Number of replicates (at least 3)
— Substrate or media characteristics (including pH) and method of preparation of
media
— Measured or nominal concentrations
— Number of plants and fronds/vessel, or cells/mi/vessel at the beginning of test
— Method used to assign test organisms to test and control groups
— Description of growth chamber
— Environmental conditions, ie. thermoperiod, relative humidity, lighting regime
(intensity, quality, and photopenod).
Method of statistical analysis.
— Identification of protocol followed (or include copy).
DISCUSSION AND RESULTS
8. — Detrimental effect of treatment compared with control, reported in ppm. Raw data provided
9. — Phytotoxicity symptoms observed.
10. — Number of plants and fronds/vessel, or cells/mi/vessel at conclusion of test as compared to
the control group.
11. — Deviations from EPA Subdivision J Guidelines and protocol.
12. Conclusions reached based on the results of the study.
13. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for deviations from the protocol
Criteria marked with a are supplemental and may not be required for every study.
C-137
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Subdivision .1
Guideline Ref. No. 122 2
Decembet 24, 1989
122-2 Growth and Reprodualon of Aquatic Plants — Tier I
GUIDANCE FOR SUMMARIZING STUDIES
Items in snmm y should inciwle the items d cmsed in Cbapter2of ski p** ’ge and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summanzed, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test organisms, including
- Common and scientific names of test organisms
3. Description of the test vessels, including:
• Test vessel type, size and shape.
4. Description of the test system, including
- Criteria used to determine effects (i.e. phytotoxicity rating system)
5. Description of the test design, including
• Test duration and observation periods
• Dose rate in ppm
DISCUSSION AND RESULTS
6. Detrimental eftect compared to control reported in ppm
7. Phytotoxicny symptoms observed
8. Number of ptants and fronds /vessel, or cells/mi/vessel at conclusion of test as compared to the
control group
9. ConcLusions reached based on the results of the study
C-138
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Subdivision J
Guideline Ref. No. 12.3.1(a)
December 24, 1989
123-1(a) Seed GerminationiSeedling Emergence — Tier II
ACCEPTANCE CRiTERIA
Does }VUZ study meet the &)IIowing a ptanee aiteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2 Identification of test substance
Source, purity, identification number, percent active ingredient
Identity and composition of impunties
Water solubility of test material reported in ppm
— Vapor pressure of test material
3. Preparation of test solution (control/treatment) described
— Solvent used and amount used to dissolve test material
If used, amount of solvent added to control
Water source and description
* Total amount of test matenal used
* Temperature of water reported
4 Test Plants
Six dictyledoneae species from at least four families and four monocotyledoneae
.pecies from at least two families are required Corn, soybean, and a root crop must
be tested. Al a minimum, test those plant species of Tier I that exhibited phytotoxic
effects.
— Family, genus, species, and cultivar or variety
Source
5. Composition and size of test vesseLs
— Test vessels type, size, and shape
— Depth of medium in relation to test vessel
6. Test System described
Criteria used to determine effects (i.e. phytotoxicity rating system)
Criteria marked with a • are supplemental and may not be required for every study.
C-139
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Subdivision J
Guideline Ref. No. 123-1(a)
December 24, 1989
7. Test design followed
Test duration and observation periods live days for seed germination, and two weeks
for seedling emergence
Dose rates in ppm (seed germination) and pounds of Al per acre (seedling
emergence), at least five doses should be tested in a geometric progression including
a no-effect level, a subtoxic level (
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Subdivision J
Guideline Ref. No. 1 2 3-1(a)
December 24, 1989
1Z3-1(a) Seed Germinationiseeilung Emergence — fler El
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should indude the items d cussed in Chapter 2 of this pa*age and the specific items
listed bekw.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of test species, including
- Family, genus, species, and cultivar or variety names
3. Descnption of test vessels, including
• Test vessel type, size, and shape
4 Description of test system, including
- Criteria used to determine effects (i.e. phytotoxicity rating system)
5. Description of test design, including
• Test duration and observation periods
Dose rates in ppm (seed germination) and pounds of Al per acre (seedling emergence)
Number of treatment levels
• Date of planting
DISCUSSION AND RESULTS
8 Phytotoxicity symptoms observed
9. Percent germination and percent emergence at the conclusion of test as compared to controls
10. EC25 and EC5O values in ppm and pounds of Al per acre with 95% confidence limits
11. The no-observed-effect level
12. Discussion of eencentration(s) producing phytotoxicity
13. Conclusions reached based on the results of the study
C-141
-------
Subdivision J
Guideline Ref. No. 123-1(b)
December 24, 1989
123-1(b) Vegetative Vigor - Tier 11
ACCEPTANCE CRiTERIA
Does ur study meet the following ptanee criteria?
GENERAL INFORMATION
1. — Date of test initiation and termination reported
MATERIALS/METHODS
2. Identification of test substance
Source, punty, identification number, percent active ingredient
— Identity and compostion of impunties
— Water solubility of test material reported in ppm
Vapor pressure of test material
3. Preparation of test solution (control/treatment) described
— Solvent used and amount used to dissolve test matenal
— If used, amount of solvent added to control
Water source and description
* Total amount of test material used
‘.. Temperature of water reported
4 Test Plants
— Six dicotyledoneae species from at least four families and four monocotyledoneae
species from at least two families are required. Corn, soybean, and a root crop must
be tested.
Family, genus, species, and cultivar, or variety
Source
5. Composition and size of lest vessels
— Test vessel type, size, and shape
— Depth of medium in relation to test vessel
6 Test system described
— Criteria used to determine effects (i.e. phyiotoxicity rating system)
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-142
-------
Subdivision J
Guideline Ref. No. 123.1(b)
December 24, 1989
7. Test design followed
— Test duration and observation periods (at least 2-weeks with weekly observations)
Dose rates in pounds of Al per acre; at least live doses should be tested in a
geometnc progression including a no-effect level, a subtoxic level (
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Subdivision J
Guideline Ref. No. 123-1(b)
December 24, 1989
123-1(b) Vegetative Vigor - Tier II
GUIDANCE FOR SUMMARIZING STUDIES
Items in snmm ry should ladude the items d c &ssed in Chapter 2 of th package and the sp fic items
listed bekjw.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test organisms, including
- Family, genus, species, and cultivar or vanety names
3. Descnption of the composition and size of test vessels,including
- Test vessel type, size, and shape
4. Descnption of the test system, including
- Critena used to determine effects (i.e. phytotoxicity rating system)
5. Description of test design followed
- Test duration and observation periods
• Dose rates in ppm and pounds of Al per acre
- Number of treatment levels
- Date of planting
DISCUSSION AND RESULTS
8. Phytotoxicity symptoms observed
9. Growth measurements taken (height and dry weight)
10. EC25 and EC5O values in ppm and pounds of Al per acre with 95% confidence limits
Ii. The no-observed-effect level
12. Discussion of ancentratlon(s) producing phytotoxicicy
13. Conclusions reached based on the results of the study
C-144
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Subdivision J
Guideline Ref. No. 123-2
December 24, 1989
123-2 Growth and Repmducrion of Aquatic Plants — 11cr II
ACCEPTANCE CRITERIA
Does your study m 1 the following ptaace criteria?
GENERAL INFORMATION
1. — Data of test initiation and termination reported.
MATERIALS/METHODS
2. Identification of test substance
— Source, purity, identification number, percent active ingredient
‘_ Identity and composition of impurities
— Water solubility of test material reported in ppm
3. Preparation of test solution (controlltreatment) described
— Solvent used and amount used to dissolve test material
— If used, amount of solvent added to control
Water source and description
Total amount of test material used
_ Temperature of water reported
4 Test Plants
— Green alga Selenastrum capncornutum , blue-green alga Anabaena flos-aguae ,
duckweed Lemna ybba , marine diatom Skeletonema costatum , and a freshwater
diatom. Include culture type and strain.
— History of test organisms described
— Source and acclimation described
5. Composition and size of test vessels described
— Test vessel type, size, and shape
— Ratio of test vessel size to medium
6. Test system described
— Criteria used to determine effects (i e. phytotoxicity rating system)
Criteria marked with a * are supplemental and may not be required for every study.
C-145
-------
Subdivision J
Guideline Ref. No. 123-2
December 24, 1989
7. Test design followed
Test duration and observation penods (algae tests at least 5 days with daily
observations, Lemna test at least 14 days with 3 day observations)
Method of introducing the test solution into the test vessels
Dose rates in ppm
Number of treatment levels (at least 5); in geometric progression. Doses should
include a subtoxic (cEC5O) and a non-toxic concentration. The highest dose should
be less than the maximum label rate.
Type and number of controls
Number of replicates (at least 3)
Substrate or media characteristic (including pH) and method of preparation of
media
— Measured or nominal concentrations
— Number of plants and fronds/vessel, or cells/mi/vessel at the beginning of test
Method used to assign test organisms to test and control groups
— Description of growth chamber
Environmental conditions, te. thermoperiod, relative humidity, lighting regime
(intensity, quality, and photopenod).
— Method of statistical analysis.
— Identification of protocol followed (or include copy).
DISCUSSION AND RESULTS
8. — EC5O values reported in ppm with 95% confidence limits. Raw data provided.
9 The no-observed-effect level determined.
10. Phytotoxictty symptoms observed.
ii. — Number of plants and fronds/vessel, or cells/mi/vessel at conclusion of test as compared to
the control group.
12. Deviations from EPA Subdivision J Guidelines and protocol.
13. — Discussion of concentration(s) producing phytoto,cicity provided.
14. — Conclusions reached based on the results of the study.
15. The relati hip between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for deviations from the protocol
Criteria marked with a ‘ are supplemental and may not be required for eveiy study.
C.146
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Subdivision J
Guideline Ref. No. 123.2
December 24, 1989
123-2 Or th and Reproduction Of Aquatic Plants - Tier II
GUIDANCE FOR SUMMARIZING STUDIES
hems in summary should Indude the items disci&ssed in Chapter 2 of th package and the specific items
listed bekiw.
GENERAL INFORMATION
1. Conclusions summarized, including date of test initiation and termination.
MATERIALS/METHODS
3. Description of the test organisms, including
• Common and scientific names of test organisms
4. Description of the composition and size of test vessels
5. Description of the test system, including
- Criteria used to determine effects (i.e. phytotoxicity rating system)
6. Description of the test design. including
- Test duration and observation periods
- Dose rates in ppm
Number of treatment levels (at least five)
DISCUSSION AND RESULTS
8. EC5O values reported in ppm with 95% confidence limits
9. The no-observed-effect level determined
10. Phytotoxicity symptoms observed
11. Number of plants and fronds/vessel, or cells/mI/vessel at conclusion of test as compared to the
control group
12. Discussion of concentration(s) producing phytotoxicicy provided
13. Conclusions reached based on the results of the study
14. Discussion relating the results of the study to test chemical
C-147
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Subdivision 3
Guideline Ref. No. 124-i
December 24, 1989
124-1 Terrestrial Field Test - Tier 111
ACCEPTANCE CRITERIA
Does your study m t the fOllowing ptan aiteria?
1. The protocol was approved by the Agency.
2. — The following documents should be used for guidance in designing and conducting a field
test:
Non-Target Plants: Terrestrial Field Testing, Tier 3, EPA 54019-86-135, June 1986.
NTIS No. PB 87-101697.
Pesticide Assessment Guidelines Subdivision J Hazard Evaluation: Nontarget Plants,
EPA 540i09- 8 2-0 2 O, October 1982. NTIS No, PB83-153940.
These references are available from the National Technical Information Service (NTIS). Orders may be
placed to NTIS by telephone at (703) 4.87-4650 or by mail to the following address:
National Technical Information Service
A1TN: Order Desk
52.85 Port Royal Road
Springfield, Virginia 22161
Criteria marked with a are supplemental and may not be required for every study.
C148
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Subdivision J
Guideline Ref. No. 124-1
December 24, 1989
124-1 Terrestrial Field Test - Tier III
GUIDANCE FOR SUMMARIZING STUDIES
Items in snmn’ ry should indude the items dL cusscd in Chapter 2 of thia package and the specific items
Listed below.
1. INTRODUCTION
- Objective of Field Studies
- General Approach
- Sampling and Experimental Design
2. STUDY
• Objective and Scope
- Geographic Area Selection
- Study Site Selection
- Number of Sites
- Size of Study Sites
• Chemical Application
- Methods
- Interpretation of Results
C.149
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Subdivision J
Guideline Ref. No. 124-2
December 24, 1989
124-2 AquatIc Field Test - Tier ifi
ACCEPTANCE CRITERIA
Does your study m t the (ollo ing a pLan aiteria?
1. — The protocol was approved by the Agency.
2. The following documents should be used for guidance in designing and conducting a field
test:
Non-Target Plants: Aquatic Field Testing, Tier 3, EPA 540 V9-86-136, June 1986.
NTIS No. PB 87-101705.
Pesticide Assessment Guidelines Subdivision J Hazard Evaluation: Nontarget Plants,
EPA 540i09-82-020, October 1982. NTIS No, P883-153940.
These references are available from the National Technical Information Service (NTIS). Orders may be
placed to NTIS by telephone at (703) 487-4650 or by mail to the following address:
National Technical Information Service
A1TN: Order Desk
5285 Port Royal Road
Springfield, Virginia 22161
Criteria marked with a * are supplemental and may not be required for every study.
C-ISO
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Subdivi io J
Guideline Ref. No 124-2
December 24, 1989
124-2 Aquatic Field Test - Tier Ill
GIJ [ DANCE FOR SUMMARIZING STUDIES
Items in snmm ry should include the items discussed in Chapter 2 of this package and the specthc items
listed below.
1 INTRODUCTION
- Objective of Field Studies
- General Approach
• Sampling and Expenmental Design
2. STUDY
- Objective and Scope
- Geographic Area Selection
- Study Site Selection
- Number of Sites
- Size of Study Sites
- Chemical Application
- Method.s
- Interpretation of Results
C- 151
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C-152
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SUBDIVISION K
132-1 Residue Dissipation Data . 154
133-3, 133-4 Dermal and Inhalation Exposure Monitonng 156
C-153
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Subdivision K
Guideline Ref. No 132.1
December 24, 1989
132-1 Residue D sipation Data
ACCEP’IANCE CRiTERIA
Does your study meet the following a ptanee afteria?
1. Typical end-use product of the active ingredient tested.
2. — Site(s) tested representative of reasonable worst-case climatic conditions expected in intended
use areas.
3. End-use product applied by application method recommended for the crop. Application rate
given and should be at the least dilution and highest, label permitted, application rate.
4 Application(s) occurred at time of season that the end-use product is normally applied to
achieve intended pest control.
5. Meteorological conditions including temperature, wind speed, daily rainfall, and humidity
provided for the duration of the study.
6. Duplicate foliar and/or soil samples collected at each collection period.
7. Sufficent collection times to establish dissipation curve. First sample time taken as soon as
sprays dry or dusts settle. Short durations should exist between earlier sample intervals and
may lengthen with later samples.
8. Control and baseline foliar or soil samples collected.
9. Residue storage stability, method efficencv (residue recovery), and limit of quantification
provided.
10 Foliar residue data expressed as /.ig or mg/cm 2 leaf surface area.
11.__ Soil residue data expressed as Lg/g of fine soil material.
12.__ Reported residue dissipation data in conjunction with toxicity data must be sufficient to
support the determination of a reentry interval.
Criteria marked with a are supplemental and may not be required for every study.
C-154
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Subdivision K
Guideline Ref. No. 132.1
December 24, 1989
132-1 Re id e Dissipation Data
GUIDANCE FOR SUMMARIZING I1JDIES
Items in summary should indude the items disc &ssed in (lapter2 of this package and the speeific items
listed bek,w.
1. Form of pesticide tested.
2a. Site(s) tested and time of season of application(s).
2b. Description of why site(s) tested are representative of geographic and climatic conditions expected to
yield reasonable worst case exposure.
2c. Meteorologic conditions for duration of study.
3. Product handled and applied.
a. equipment
b. application rates
c. number of applications
d. interval between applications (if answer to is 2 or greater)
4. Description of collection techniques for foliar/soil samples.
5. Sample collection times.
6. QAIQC data including storage stability, method efficiency, field recovery, and limit of quantification
provided.
7. Reporting of foliar and/or soil residue data corrected for field recovery (less than 100%).
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Subdivision K
Guideline Ref. No. 133-3, 133.4
December 24, 1989
133-3, 133-4 Dernial and InhAbtio n Exposure Monitoring
ACCEPTA)JCE CRITERIA
Does your study meet the following a pLanee aiteria?
1. — Typical end-use product of the active ingredient tested.
2. — End-use product applied at maximum allowable rate using equipment recommended for that
end-use product on the crop.
3 — Dermal and inhalation exposure monitored by validated methodologies (i.e. patches, whole
body dosimeters, personal air samplers), as appropriate.
4. — Study participant activity contributing to exposure and clothing should be consistent with
typical accepted agricultural practices.
5. Duration of sampling is sufficent to collect measurable residues but not excessive so that
residue loss occurs.
6. Each sampling period should use at least 10 workers.
7. Dermal exposure for each body area monitored for each study participant.
8. Total dermal and inhalation exposure reported for each individual and for the group as a
whole, as appropriate.
9 Storage stability, method efficency for each collection matrix, and the limit of quantification
provided.
10. Concurrent foliar residue data (jig or mg/cm 2 ) and soil residue data (/Lg/g fine soil material)
collected as per Subdivision K section 132, as appropnate.
11 — Brief description of activities including: nature of human activity, principle source of
exposure, typical environmental conditions, level of exertion and expected frequency and
duration of monitored activity.
Criteria marked with a * are supplemental and may not be required for every study.
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Subdivision K
Guideline Ref. No. 133.3, 133-4
December 24, 1989
133-3, 133-4 Dernial and lnh2bltion Exposure Monitoring
GUIDANCE FOR SUMMARIZING SrIJDIES
Items in snmm ry should indude the items d sed in Chapter 2 of th package and the specific items
Listed below.
1. Form of pesticide tested.
2. Product handled and applied.
a. equipment
b. application rates
c. number of applications
d. interval between applications (if answer to “c is 2 or greater)
3. a. Activity of study participants.
b. Clothing scenario studied for all participants.
4. Duration of monitoring period.
5. Number of workers (replicates) monitored.
6. Method for monitonng dermal and/or inhalation exposure
7. QA/QC data including storage stability, method efficiency field recovery for each collection matrix,
and the limit of quantification provided.
8. Reporting of dermal (by patch location, body part and whole body) and/or inhalation exposure,
corrected for field recovery (less than 100%).
C- 157
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C- 158
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SUBDIVISION L
141-1 Acute Toxicity Test for Honey Bees . 160
14 1-2 Residual Toxicity Test for Honey Bees 163
141-5 Field Testing for Pollinators
C-159
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Subdivision L
Guideline Ref. No. 141-1
December 24, 1989
141-1 Acute Toxiaty Test for Honey Bees
ACCEPTANCE CRiTERIA
Does mur study meet the ft)llowing a ptance a iteria?
GENERAL INFORMATION
1. _____ Date of test initiation and termination reported
MATERIALS/METHODS
2. Preparation of test solution (control/treatment) described
- Any vehicle used to dissolve test material
_____ If used, amount of vehicle added to control
*_____ Method used to get test material into solution was descnbecl
•_____ Water solubility of test matenal reported in ppm
* Total amount of test material used
3. Test Insects
Testing was conducted on the honey bee, Apis mellifera
•_____ History of test organisms described (strain, diseases and treatment)
•_____ Health described (sickness; Injuries; abnormalities; name of medication, if used.
pretest diet)
4. Test cages described
_____ Size and composition of test cages
5. Test system described
_____ Procedures used to prepare toxicant stock solution
_____ Criteria used to determine effects
6. Test design followed
•_____ Method used in assigning test bees to test and control groups and number of
replicates used
_____ Number of bees per treatment level and control
____ Number of treatment levels
*_____ Method used to determine treatment levels
— Number of bees per cage
____ Number and type of controls
Environmental conditions (ambient temperature, humidity)
Source and availability of food and water
Length of total observation period
Frequency of each observation
Cntena marked with a are supplemental and may not be required for every study.
C- 160
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Subdivision L
Guideline Ref. No, 141.1
December 24, 1989
DISCUSSION AND RESULTS
7. ____ LD5O value reported in micrograms per bee with 95% confidence limits. (Graphs, pnntouts,
and other calculations are optional). Raw mortality data provided.
8. ____ Detailed description of any unusual symptoms which occur.
9. ____ Statistical method referenced
10 The relationship between the results of the study and the test chemical described, including
_____ Obvious problems that occurred during the Study as well as the effect, if any, these
problems had on the outcome of the study
_____ Rationale for minor deviations from the protocol
Criteria marked with a are supplemental and may not be required for every study.
C.161
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Subdivision L
Guideline Ref. No. 141-I
December 24, 1989
141-1 Acute Toxaty Tess for Honey Bees
GUIDANCE FOR SUMMARIZING S11JDIES
Items in summary should include the items d cmsed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test Initiation and termination.
MATERIALS/METHODS
2. Description of the test solution (control/treatment) preparation, including
- Any vehicle used to dissolve test matenal
- Amount of vehicle added to control, if used
3. Description of the test insects, including
- Common and scientific names
4. Description of the test cages, including
5 Descnption of the test system, including
- Criteria used to determine effects
6 Description of the test design, including
- - Number of bees per treatment level and control
• Number of treatment levels
- Number of bees per cage
- Number and type of controls
DISCUSSION AND RESULTS
7. LD5O value reported in micrograms per bee with 95% confidence limits. (Graphs, printouts, and
other calculations should be attached to report). Raw mortality data provided.
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Subdwision L
Guideline Ret. No. 141-2
December 24, 1989
141-2 Residual Toxidty Test for Honey Bees
ACCEPTANCE CRITERIA
Does ur study meet the following a ptanee afteria?
GENERAL INFORMATION
1. _____ Dates of test initiation and termination reported
MATERIALS/METhODS
2. Preparation of test solution (control/treatment) described
Any vehicle used to dissolve test material
_____ If used, any vehicle added to control
_____ Amount of test matenal used in pounds active ingredient per acre
3 Test Insects
— Testing was conducted on the honey bee, mellifera
S History of test organisms described (strain, diseases and treatment)
_ Health described (sickness; injunes; abnormalities; name of medication. if used:
pretest diet)
4. Test cages descnbed
— Size and composition of test cages
5 Test system described
— Procedures used to prepare toxicant solution
— Criteria used to determine effects
6. Test design followed
‘ Method used in assigning test bees to test and control groups and number of
replicates used
— Number of bees per treatment level and control
— Number of treatment levels
Basis for selection of treatment levels
— Number of bees per cage
— Number and type of controls
— Method of application to crop
— Name of crop treated
Critena marked with a * are supplemental and may not be required for every study.
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Subdivision L
Guideline Ref. No. 141-2
December 24, 1989
Method used to harvest treated foliage
Amount of treated foliage per cage
Environmental conditions
— Source and availability of food and water
Length of total observation penod
Frequency of observations
DISCUSSION AND RESULTS
7. _... ..Percent mortality of test bees reported at each treatment level and time interval. (Graphs,
printouts, and other calculations are optional). Raw mortality data provided.
8. ... ... . . . . . . .Detailed description of any unusual Symptoms which occur.
9. The relationship between the results of the study and the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the Study
Rationale for minor deviations from the protocol
Criteria marked with a are supplemental and may not be required for every Study.
C-164
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Subdivision L
Guideline Ref. No. 141-2
December 24, 1989
141-2 Residual Tox caty Test (or Honey Bees
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should indude the items diccussed in Chapter 2 of this package and the specific items
listed below.
GENERAL INFORMATION
1. Conclusions summarized, including date of test Initiation and termination.
MATERIALS/METHODS
2. Description of the preparation of the test solution (control/treatment), including
• Test material used in pounds active ingredient per acre
3. Description of the test insects, including
- Common and scientific names
4. Description of the test cages, including
• Size and composition of test cages
5. Description of the test system, including
- Criteria used to determine effects
6. Description of the test design, including
- Number of bees per treatment level and control
- Number of treatment levels
- Number of bees per cage
- Number and type of controls
- Method of application to crop
• Name of crop treated
DISCUSSION AND RESULTS
7. Percent mortality of test bees reported at each treatment level and time interval. (Graphs, printouts,
and other calculation should be attached to report). Raw mortality data provided.
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Subdivision L
Guideline Ref. No. 141-5
December 24, 1989
141-5 Field Testhig for Pollinators
ACCEPTANCE CRiTERIA
Does your study meet the Ibilowing a ptance aiteria?
GENERAL INFORMATION
_____ Dates of test initiation and termination reponed
MATERIALS/METHODS
2. Preparation of test solution (control/treatment) described
Any vehicle used to dissolve or dilute the test substance
Description of preparation methods
3. Test Insects
Identify test insect to species
— Identify source of supply of test bees
History of test organisms described (strain, diseases and treatment)
_ Health described (sickness; injuries, abnormalities; name of medication, if used;
pretest diet)
4. Dosing method described
— Amount of test material (per acre, per colony, etc.
— Method of administration
— Rationale for selection of method, route, or frequency
5. Test design followed (Note: Actual criteria would be determined through pre-test consultation with
the Agency.)
— Number of treatment levels
— Number of bees per treatment level
‘_ Method of assigning bees to treatment and control groups
Method used to determine treatment levels
— Size of eages or colonies, if appropnate
Number of bees per cage or colony
Number of controls
Type of controls
— Environmental conditions, such as ambient temperature, humidity, weather conditions
— Source and availability of food and water
— Length of total observation period
— Frequency and duration of each observation
Define criteria for determining effects
Criteria marked with a are supplemental and may not be required for every study.
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Subdivision L
Guideline Ref. No. 141-5
December 24, 1989
DISCUSSION AND RESULTS
6._ Observed adverse effects on bees/bee colonies due to treatments. (Graphs, printouts, and
other calculations are optional).
7. — Detailed description of the nature, incidence, time of occurrence, severity and duration of all
observed toxic effects, including death and any other abnormal or unusual signs.
8. — Indication of test procedures which deviated from those agreed upon during pre-test
consultation with the Agency. Provide a rationale for the changes.
9 — Statistical methods referenced
10. The relationship between the results of the study arid the test chemical described, including
— Obvious problems that occurred during the study as well as the effect, if any, these
problems had on the outcome of the study
— Rationale for minor deviations from the protocol
Criteria marked with a * are supplemental and may not be required for every study.
C-167
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Subdivision L
Guideline Ref No. 141-5
December 24, 1989
141-5 Field Testing (or Pollinators
GUIDANCE FOR SUMMARIZING STtJDEES
Items in snnim ry should üidude the items d a sed in Chapter 2 of th package and the specific items
listed below.
GENERAL ENFORMATION
1. Conclusions summarized 1 including date of test initiation and termination.
MATERIALS/METHODS
2. Description of the test insects, including
• Common and scientific names
3. Description of the dosing method, including
- Amount of test matenal (per acre, per colony, etc.)
Method of administrations
4 Description of the test design, including
- Number of treatment levels
- Number of bees per treatment level
- Size of cages or colonies, if appropnate
- Number of bees per cage or colony
- Number of controls
- Type of controls
- Define cricefia for determining effects
DISCUSSION AND RESULTS
5 Observed adverse effects on bees/bee colonies due to treatments. Optional graphs, pnntouls, and
other calculation may be attached to summaty. Raw mortality data provided.
6. Detailed description of the nature, incidence, time of occurrence, severity, and duration of all
observed toxic effects, including death and any other abnormal or unusual signs.
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SUBDIVISION M
MICROBIALs
151A.10 Product Identity . 172
1S1A-11 Manufacturing 174
151A-12 Discussion of Formation of Unintended Ingredients 175
1SIA-13 Analysis of Samples 176
151A-15 Certification of Limits 177
151A-16(a-j) Physical and Chemical Properties 178
152A-1O Acute Oral Toxicity/Pathogenicity iso
152A-11 Acute Dermal Toxicity 181
152A.12 Acute Pulmonaiy Toxicity/Pathogenicity 182
152A-13 Acute Intravenous ToxicitylPathogenicity 183
152A.14 Primary Eye Irritation Study 184
152A.1S Hypersensitivity Incidents 185
152A-16 Cell Culture 186
152A-20 Acute Toxicity 187
152A-21 Subchronic Toxicity/Pathogenicity . . 188
152A-30 Reproductive/Fertility Effects 189
152A-31 Oncogenicity [ 90
152A-32 Immunodeficiency . . i i
152A-33 Primate InfectwityfPathogenicity 192
153A-4 through 153A-14 Residue Chemistry 193
154A-16 Acute Avian Oral Test 194
154A-17 Avian Inhalation Test 195
154A-18 Wild Mammal Test 1%
154A-19 Freshwater Fish Test 197
154A-20 Freshwater Aquatic Invertebrate Test 198
154A-21 Estuarine and Manne Animal Test (Estuanne Fish) . . 199
154A-21 Estuanne and Marine Animal Test (Estuanne Shrimp) . 200
154A-22 Noncarget Plant Studies . 201
154A-23 Nontarger Insect Test 202
154A-24 Honey Bee Test 203
154A-25 Terrestrial Wildlife & Aquatic Organism Testing 204
154A.26 Avian Chronic Pathogenicity and Reproduction Test 205
154A-27 Aquatic Invertebrate Range Test . 206
154A-2.8 Fish Life Cycle Studies 207
154A-29 Aquatic Ecosystem Test 208
154A-31 Nontarget Plant Studies 209
154A-33 Simulated and Actual Field Tests (Birds, Mammals) 210
154A-34 Simulated and Actual Field Tests (Aquatic Organisms) . 211
154A-35 Simulated and Actual Field Tests (Insect Predators, Parasites) . 212
154A.36 Simulated and Actual Field Tests (Insect Pollinators) 213
155A .1O through 155A-12 Environmental Expression 214
C-169
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BIOCHEMICALS
1518-10 Product Identity . 215
1513-11 Manufacturing Process 216
151342 Discussion of Formation of Unintended Ingredients 217
1518-13 Analysis of Samples 218
151B-15 Certification of Limits 219
151B-17 Physical and Chemical Properties 220
152B-10 Acute Oral Toxicity 223
152B-11 Acute Dermal Toxicity 224
152B-12 Acute Inhalation Toxicity 225
152B-13 Primary Eye Irritation 226
152B-14 Primary Dermal Irritation 227
152B-15 Hypersensitivity 228
152B-17 Genotoxicity 229
152B .18 lmmunotoxicity 230
152B-19 Mutagenicity 231
152B-20 Subchromc Feeding 232
ISZB.21. Repeated Dermal Dose Toxicity (90-day) 234
152B.22 Subchronic Inhalation Toxicity (90-day) 236
152B.23 Teratology 23.8
1528-24 Immunotoxicity -- Tier 11 239
152B-26 Chronic Feeding 240
152B -29 Oncogenicity 242
1538.3(a) Chemical Identity 243
1538.3(b) Directions for Use 244
1533.3(c) Nature of the Residue-Plants 246
153B-3(d) Nature of the Residue-Animals 247
153B.3(e and t) Residue Analytical Method 248
153B-3(g) Storage Stability 249
153B.3(h) Magnitude of the Residue -- Potable Water 250
1538-3(i) Magnitude of the Residue - - Fish 251
153B-3(j) Magnitude of the Residue -- Errigated Crops 252
153B-3(k) Magnitude of the Residue-- Food Handling 253
1538-3(1) Magnitude of the Residue -- Meat, Milk, Poultry, Eggs 254
153B-3(m) Reduction of Residues 255
153B-3(n) Magnitude of the Residue - - Crop Field Trials 256
153B-3(o) Magnitude of the Residue -- Processed Food 257
154B-6 Acute Avian Oral Test 258
1548-7 Avian Dietary Test 259
154B-8 Freshwater Fish LC Test - 260
154B-9 Freshwater Aquatic Invertebrate LC Test 261
154B-10 Nontarget Plant Studies (Seed Germination/Seedling Emergence) 262
1548-10 Noniarget Plant Studies (Vegetative Vigor) 263
154B-11 Nontarget Insect Testing 264
154B42 Terrestrial Wildlife Testing 265
154B-13 Aquatic Animal Testing 266
154B-14 Nontarget Plant Studies 267
154B-15 Nontarget Insect Testing 268
155B-4 Volatility Study (Lab) 269
155B-4 Volatility Study (Field) 270
C-170
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155B.5 & 155B-6 LeaChing/AdsorptionjDesorption
155B-8 Ultraviolet Absorption Study
155B-9 Hydrolysis Study
155B-1O Aerobic Soil Metabolism Study
155B-1i Aerobic Aquatic Metabolism Study
155B-12 Photolysis in Soil Study
155B-13 Photolysis in Water Study
271
273
274
275
276
277
278
C- 171
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Subdivision M
Guideline Ref. No. 151A-10
December 24, 1989
151A-1O Product Identity
ACCEPTANCE CRITERIA
Does your study meet the tbllowing a ptan aiteria?
1. — Product name and trade name (if different)
2. — Name, nominal concentration (for microbial ingredients, the amount present in the product
in recognized units of potency, percentage of weight, units of microbial pesticide per unit
weight or volume of product, or other appropriate expression of biological activity, PFU,
CFU, etc.), and certified limits (upper and lower) for each active ingredient and each
intentionally-added inert ingredient
3. — Name and upper certified limit for each impurity or each group of impurities present at >
0.1% by weight and for certain toxicologically significant impurities (e.g., microbial toxins)
present at < 0.1%
4. Identity and purpose of each active ingredient and each intentionally-added inert
5. Taxonomic description of all living microorganisms in the product, methodologies and results
of all tests performed in support of that classification, and (optional) any company assigned
experimental or internal code numbers for each active microbial ingredient.
The common, alternative, and superseded names
The natural occurrence of the organism, its relationship to other species (particularly
those that are pathogenic), and its history
The biological properties of the active agent with respect to target species, pest host
range, life cycle, and mode of action. With respect to the properties of the microbial
agent, any known or potential hazard (such as infectivity) to mammals (including
humans), the environment, and nontarget species should be discussed.
— A description of the morphological types of the microbial pesticide and any unusual
morphological, biochemical, or resistance characteristic(s) of the organism if such
characteristic(s) are different from the classic description of the organism.
— If the microbial pesticide contains plasmids or other extrachromosomal genetic
elements involved in pesticidal activity, pathogenicity, toxicity, etc., these must be
identified as well as any known phenotypic characters coded by such elements and
their stability.
6. Chemical name front Chemical Abstracts Index of Nomenclature and Chemical Abstracts
Service (CAS), Registry Number for each chemical active ingredient and, if available, for
each intentionally-added inert
7 Molecular, structural, and empirical formulas, molecular weight or weight range, and any
company assigned experimental or internal code numbers for each active chemical ingredient
8 — Desalpilon of each beginning material in the manufacturing process for chemical ingredients
— EPA Registrauon Number if registered; for other beginning materials, the following:
Name and address of manufacturer or supplier
— Brand name, trade name or commercial designation
Technical specifications or data sheets by which manufacturer or supplier describes
composition, properties or toxicity
Criteria marked with a are supplemental and may not be required for every study.
C-172
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Subdivision M
Guideline Ref. No. 15 lA-tO
December 24, 1989
9. — Culture allection from which the microbial active ingredient may be obtained.
10. — Similarity of currently-utilized strains to those originally registered or used as test strains for
hazard testing, i.e. are they identical and, if not, provide strain designations and any known
differences.
11. — An updated Confidential Statement of Formula (EPA Form 8570-4, rev. 9i 7)
Criteria marked with a • are supplemental and may not be required for every study.
C-173
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Subdivision M
Guideline Ref. No. 15 lA-Il
December 24, 1989
151A-I1 Manufacturing Process
ACCEPTANCE CR ERlA
Does )VU( study meet the following a planee crneria?
1. — Description of manufacturing/fermentation process
2. — Statement of whether batch or continuous process
3. — Relative amounts of beginning and Intermediate materials, including EPA Registration
numbers, if any, source and punty of materials, and order in which they are added
4 — Description of equipment
5. — Description of physical conditions (temperature, pH, pressure, humidity) controlled in each
step and the parameters that are maintained
6. Description of steps taken to limit extraneous contamination both chemical and biological
7. Description of the procedures used to establish the identity and purity of the culture from
which the unformulated microbial pesticide is produced
8. Duration of each step of process
9 — Description of punfication procedures
10. Description of the techniques used to ensure a uniform or standardized product.
11. — A clear presentation of the stage at which inerts are intentionally added, if and when any
concentration is effected, the material to be used as the manufacturing use product (MP),
whether MP registration is sought, and wheiher a TGAI/MP is sold and/or shipped
Criteria marked with a are supplemental and may not be required for every study.
C-174
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Subdivision M
Guideline Ref. No. 151A-12
December 24, 1989
151A-12 Discussion of Formation of Unintended ngredients
ACCEPTANCE CRfl’ERIA
Does your study m 1 the ibliowing a ptan criteria?
Discussion of formation of impurities based on established chemical and microbial theory
addressing (1) each impurity which may be present at O.1% or was found at O.1% by
product analyses and (2) any extraneous material that might reasonably be present in the
pesticide product, e.g. allergens, microbial toxins and other metabolic products, mutant
strains, microbial contaminants with particular reference to potentially infective or
antagonistic forms, side products from chemical reactions employed in the manufacturing
process, fermentation residues from the growth of bacteria or fungi, extraneous host residues
from viruses produced in cell cultures, whole animals, or other living forms, residues of
contaminants that remain following the purification or extraction process, and impurities in
chemicals used in the manufactunng process.
Critena marked with a • are supplemental and may not be required for every study.
C-175
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Subdivision M
Guideline Ref. No 151A-13
December 24, 1989
151A-13 Analysis of Samples
ACCEPTANCE CRITERIA
Does vur study meet the following a planee aiteria?
1. — Five or more representative samples (batches in case of batch process) analyzed for each
active ingredient and all impurities or microbial contaminants present at O.1%
2. — Degree of accountability or closure 98%
3. — Analyses conducted for certain trace toxic impurities at lower than 0.1% (examples, certain
microbial toxins or nitrosamines in the case of products containing dinitroanilines or
containing secondary or tertiary alnines/alkanolamines plus nitrites; polyhalogenated
dibenzodioxjns and dibenzofurans) [ Note that in the case of nitrosammes both fresh and
stored samples should be analyzed.]
4. — Complete and detailed description of each step in analytical method used to analyze above
samples. A quantitative serological or other appropriate test for the microbial pesticide may
be substituted for chemical analysis.
5. — Statement of precision and accuracy of analytical method used to analyze above samples
6. — Identities and quantities (including mean and standard deviation) provided for each analyzed
ingredient
7. — The test material is to be the purest pesticidal grade commercially produced prior to
intentional addition of inerts. Generally, this test material is the same as that used for
certain nontarget and human hazard testing and is identical to, or equivalent to the technical
grade. Any differences from the test substance used for hazard testing should be noted.
Criteria marked with a • are supplemental and may not be required for every study.
C-176
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Subdivision M
Guideline Ref. No. 151A-15
December 24, 1989
15LA-15 Certification of Limits
ACCEPTANCE CRiTERIA
Does vur study meet the following a ptanee aiteria?
1. — Upper and lower certified limits proposed for each active ingredient and intentionally added
inert along with explanation of how the limits were determined. The limits for microbial
agents should be expressed as: (i) MPCA units per unit weight or volume, (ii) international
units of potency per unit weight, and (iii) weight percent of product. Items (i) and (ii) may
be determined using biological, genetic, biochemical, serological, or other appropnate tests.
2. — Upper certified limit proposed for each impurity or microbial contaminant present at
0.1% and for certain toxicologically significant impurities at < 0.1% along with explanation
of how limit determined
3 Analytical methods to ven1 ’ certified limits of each active ingredient and impurities (latter
not required if exempt from requirement of tolerance or if generally recognized as safe by
FDA) are fully described
4. Analytical methods to verify certified limits validated as to their precision and accuracy
Cnteria marked with a are supplemental and may not be required for every study.
C.177
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Subdivision M
Guideline Ref. No. l 5 IA-16(a.j)
December 24, 1989
151A-16(a-j) Physical and ( emicaJ Properties
ACCEFrM CE CRITERIA
Does your study meet the tbUowing a pIance criteria?
A. Color
— Verbal description of coloration (or lack of it)
Any intentional coloration also reported in terms of Munsell color system
B. Physical State
— Verbal description of physical state provided using terms such as ‘solid, granular,
volatile liquid’
Based on visual inspection at about 20-25°C
C. Odor
Verbal description of odor (or lack of it) using terms such as “moldy, or “vinegar-
like’, etc.
— Observed at room temperature
D. Density or Specific Gravity
Measured at about 20-25°C
Density/bulk density reported in g/mI the specific gravity of liquids reported with
reference to water at 20°C [ NOTE: For a solid in particulate form a measurement
of bulk density may be substituted for measurement of density, or for microbial
suspensions, optical density may be used if related to colony forming units per mlJ
E.pH
— Measured at about 20-25°C
If matenal is a solid, measured after dilution or dispersion in distilled water
F. Stability
Sensitivity to metal ions and metal determined
Stability at normal and elevated temperatures
Sensitivity to sunlight determined
Determination of genetic stability as related to any toxic/pathogenic properties
apressed by the MPCA
G. Storage Stability
Product stored in its commercial package or smaller one of same construction and
materials
Amount of active ingredient determined in product at beginning and end of test
period (duration of at least one year for a product which degrades sufficient
duration to support expiiation date)
Critena marked with a * are supplemental and may not be required for every study.
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Subdivision M
Guideline Ref. No. 151A-16(a-j)
December 24, 1989
— Any deterioration or degradation products determined
— Product examined for physical or biological changes, or for growth and/or production
of toxins at end of test
— Product stored at about 20-25°C (and 50% relative humidity if permeable packaging)
under warehouse conditions reflecting expected storage
Report includes duration and conditions of storage, quantitative analyses of active
ingredient, and identification of any deterioration, degradation products, or physical
changes (and consequences of latter on safe handling and use of product)
H. Viscosity
— Determined at about 20-25°C
— Reported in poises, stokes, or other conventional units
I. Miscibility
— Determined at about 20-25°C
— Product mixed with petroleum solvents whose composition reflects those on label and
at rate on label
— Mixture examined for separation after 30 minutes
J. Corrosion Characteristics
Data on corrosion characteristics provided (experimental method described)
reasonable explanation given for lack of corrosiveness based on nature of product
(e.g., lack of extreme pH; unreactive)
Criteria marked with a * are supplemental and may not be required for every study.
C-179
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Subdivision M
Guideline Ref. No. 152A-1O
December 24, 1989
152A-1O Acute Oral Toxicity/Pathogeniaty
ACCEPTANCE CRrFERIA
Does vur study meet the following a ptau criteria?
General Information
I — Raw data in support of Final Report are available
2. Date of test initiation and ternunacion reported
Materials and Methods
3. Identification of test substance
— Appropriate t.axonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Cnteria marked with a are supplemental and may not be required for evety study.
C-180
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Subdivision M
Guideline Ref. No. 152A-I1
December 24, 1989
152k .11 Acute Dermal Toxicity
ACCEPTANCE CRITERIA
Does vur study meet the following aa pIanee aiteria?
I. — Technical form of the active ingredient tested (for reregistration only).
2._ At least 5 animals, exJgroup
Rats 200-300 gm, rabbits 2.0-3.0 kg, or guinea pigs 350-450 gm
4. — Dosing, single dermaL
5. — Dosing duration at least 24 hours.
6. • Vehicle control, only if toxicity of vehicle is unknown.
7. — Doses tested, sufficient to determine a toxicity category or a limit dose (2000 mg’lcg).
8. Application site clipped or shaved at least 24 hours before dosing
9 — Application site at least 10% of body surface area.
10. Application site covered with a proous nonirritating cover to retain test material and to
prevent ingestion.
11. — Individual observations (at least once a day).
12. Observation period to last at least 14 days, or until all test animals appear normal, whichever
is longer.
13 — Individual daily observations
14 Individual body weights
15._ Gross necropsy on all animals.
Criteria marked with a are supplemental and may not be required for every study.
C-181
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Subdivision M
Guideline Ref. No. 152A- 12
December 24, 1989
152A-12 Acute Pulmonaiy Toxzdty/Pathogeiuaty
A(XEPTANCE CRITERIA
Do your study mccl the fbllowing a plancc aitena?
General Information
1. Raw data in support of Final Report are available
2. — Date of t t initiation and termination reported
Materials and Methods
3. Identification of test substance
— Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Criteria marked with a • arc supplemental and may not be required for every study.
C-182
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Subdjvisjo M
Guideline Ref. No. 152A-13
December 24, 1989
152A-13 Acute IntrawnoiLs Toxicity /P hogeniaty
A PTANCE CRflERIA
Does )VUZ study meet the foL owing a ptancc aiteria?
General Information
1. — Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Materials and Methods
3. Identification of test substance
— Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Criteria marked with a are supplemental and may not be required for every study.
C-183
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Subdivision M
Guideline Ref. No. 152A-14
December 24, 1989
152A-14 Primaiy Eye Irritation Study
ACCEPTANCE CRflERIA
Do ,vur study meet the following aa ptance aiteria?
1. Technical form of the active ingredient tested (for reregistration only).
2. — Study not required if material is corrosive, causes severe dermal irritation or has a pH of 2
or 11.5.
3 6 adult rabbits
4. Dosing, instillation into the conjunctival sac of one eye per animal.
5. . ..... ... . .. Dose, 0.1 ml if a liquid; 0.1 ml, or not more than 100mg if a solid, paste, or particulate
substance.
6. Solid or granular test material ground to a line dust.
7. Eyes not washed for at least 24 hours.
8. — Eyes examined and graded for irritation before dosing and at 1, 24, 48, and 72 hr, then daily
until eyes are normal or 21 days (whichever is shorter).
9. Individual daily observations.
Cnteria marked with a • are supplemental and may not be required for every study.
C- 184
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Subdivtsion M
Guideline Ref. No. 152A-15
December 24, 1989
152.A-15 Hypei ensiüvity Incidents
A(XEPTANCE CRiTERIA
Does vuz study meet the following a pIance criteria?
General Information
1. — Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Materials and Methods
3. Identification of test substance
— Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Cnteria marked with a are supplemental and may not be required for every study.
C-185
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Subdivision W
Guideline Ref. No. 152A46
December 24, 1989
152A-16 Cell Culture
ACCEPTANCE CRITERIA
Do your study meet the blowing ptance aiteria?
General Information
1. Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Materials and Methods
3. Identification of test substance
— Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Criteria marked with a * are supplemental and may not be required for every study.
C- 186
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Subdivision M
Guideline Ref. No. 152A.20
December 24, 1989
152A-20 Acute Toxicity
ACCEPTANCE CRITERIA.
Does ur study m t the following a psan aitena?
General Information
1. — Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Materials and Methods
3. Identification of test substance
Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Cnteria marked with a are supplemental and may not be required for every study.
C-187
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Subdivision M
Guideline Ref. No. 152A-21
December 24, 1989
152A-21 Subclironic Tozicity/Pathogeniaty
AQEFrANCE CRiTERIA
Does your study meet the foLlowing a ptanee aitnria?
General information
1. — Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Materials and Methods
3. Identification of test substance
— Appropriate taxononuc identification of test microorganism and relationship to
registered active ingredient as currently produced.
Critena marked with a * are supplemental and may not be required for every study.
C-188
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Subdivision M
Guideline Ref. No. 152A-30
December 24, 1989
152A-30 ReproductivelFenuuty Effects
ACCEFFANCE CRfltRIA
Does vur study meet the following a ptanee aiteria?
General Information
1. — Raw data in support of Final Report are available
2. — Date of test Initiation and termination reported
Materials and Methods
3. Identification of test substance
— Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Criteria marked with a ‘are supplemental and may not be required for every study.
C-189
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Subdivision M
Guideline Ref. No. 152A-31
December 24, 1989
152A-31 Onugenicity
ACCEPTANCE CRITERIA
Does )VU! study meet the following a p1ance criteria?
General Information
1. — Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Materials and Methods
3. Identification of test substance
Appropnate caxonomic identification óf test microorganism and relationship to
registered active ingredient as currently produced.
Critena marked with a are supplemental and may not be required for every study.
C-190
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Subdivision M
Guideline Ref. No. 152A-32
December 24, 1989
152A-32 Iinrnunodeficiency
ACCEPTANCE CRITERIA
Does )VUI Study m t the following a eptance criteria?
General Information
1. — Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Matenals and Methods
3. — Identification of t t substance
Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Criteria marked with a * are supplemental and may not be required for every study.
C-191
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Subdivision M
Guideline Ref. No. 152A.33
December 24, 1989
152A-33 Primate Infectivity/Pathogeniaty
ACCEPTA1 JCE CRiTERIA
Do pour study meet the following a ptan aiteria?
General Enformation
1. — Raw data in support of Final Report are available
2. — Date of test initiation and termination reported
Materials and Methods
3. — Identifi tjon of test substance
— Appropriate taxonomic identification of test microorganism and relationship to
registered active ingredient as currently produced.
Criteria marked with a * are supplemental and may not be required for evety study.
C-In
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Subdivision M
Guideline Ref. No. 153A .4 through 153A44
December 24, 1989
153A-4 through 153A-14 Residue aiem uy
A( TANCE CRITERIA
Does your study meet the following au ptaace criteria?
All currently registered microbial pesticides with food uses have been granted exemptions from
requirements for a tolerance. Therefore, no residue chemistry studies have been submitted in support of
registration. Since techniques for conducting residue chemistry studies are specific to the unique
characteristics of the microbial pesticide, it is not possible to set forth generic acceptance criteria at this
time. However, all studies should include basic identification data:
I — Product name and trade name (if different)
2. EPA Registration Number if registered
3 Taxononuc description of the active ingredient
Criteria marked with a * are supplemental and may not be required for every study.
C- 193
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Subdivision M
Guideline Ref. No. 154A-16
December 24, 1989
154A-16 Acute Avian Oral Test
ACCEPTANCE CRrFERJA
Does ur study meet the following a ptanee aiteria?
1. — Raw data in support of final report are available
2. — Identity of test MPCA genus, species and variety, strain history, identification criteria
3’ Type of toxins produced by strain are well defined
4. Technical grade active ingredient (TGAJ) used in test (see Subdivision M for TGAJ
definition)
5. Test bird used was mallard duck or bobwhite quail
6. The MPCA dose as recommended in Subdivision M
7. Number of birds/treatment level(s) and controls (‘10 each)
8. Type of controls listed, including solvent, carrier, or whether negative or positive
Diet and any antibioti , vitamins or food additives described
10. Observation period (‘30 days)
Result reporting:
11. Include LD value in mg/kg and the no-observed-effect level
12. Raw mortality data provided
13. Description of observed toxic effects, including death and any other abnormal signs or
behavior
14. Pathological changes as noted by gross necropsy examination described
15. Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a are supplemental and may not be required for every study.
C-194
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Subdivision M
Guideline Ref. No. 154A-17
December 24, 1989
154A-17 Avian Inhah tjo Test
ACCEPTANCE CRITERIA
Does your study meet the following a pIance aiteria?
1. — Raw data in support of final report are available
2. — Identity of test MPCA genus, species and variety, strain history, identification criteria
3.’__ Type of toxins produced by strain are well defined
4. Technical grade active ingredient (TOM) used in test (see Subdivision M for TOM
definition)
5. — Test bird used was mallard duck or bobwhite quail
6. The MPCA dose as recommended in Subdivision M
7 — Route of administration as recommended in Subdivision M
8. — Number of birds/treatment level(s) and conirols (10 each)
9. T ’pe of controls listed, including solvent, carrier, or whether negative or positive
1O._ Diet and any antibiotica, vuanuns or food additives described
11. Observation period (30 da )
Result reporting:
12. Include LD 50 value in mg/kg and the no-observed-effect level
13. Raw mortality data provided
14 — Description of observed toxic effects, including death and any other abnormal signs or
behavior
15. — Pathological changes as noted by gross necropsy examination described
16. — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a * are supplemental and may not be required for every study.
C-195
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Subdivision M
Guideline Ref. No. 154-18
December 24, 1989
154A-1S Wild Mamm2I Test
ACCEPTANCE CRITERIA
Does vur study meet the following a ptan aiteria?
1. — Raw data in support of final report are available
2. Identity of test MPCA genus, species and variety, strain history, identification criteria
3 * Type of toxins produced by strain are well defined
4 Technical grade active ingredient (TGAI) used in test (see Subdivision M for TGAJ
definition)
5 Test species common and scientific name provided
6. The MPCA dose as recommended in Subdivision M
7. Number of animals/treatment level(s) and controls (10 each)
8. — l’ype of controls listed, including solvent, carrier, or whether negative or positive
9.’__ Diet and any antibiotica, vitamins or food additives described
[ 0. — Observation period (30 days)
Result reporting:
11. — Include LD value in mg/kg and the no-observed-effect level
12. — Raw mortality data provided
13. Description of observed toxic effects, including death, weight loss and any other abnormal
signs or behavior
14. — Pathological changes as noted by gross necropsy examination described
15. — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a * are supplemental and may not be required for every study.
C-196
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Subdivision M
Guideline Ref. No. 154A-19
December 24, 1989
154A-19 Freshwater Fish Test
ACCEPTANCE CRITERIA
Does vur study meet the following a ptance criteria?
1. — Raw data in support of final report are available
2. — Identity of test MPCA genus, species and variety, strain history, identification criteria
3 ‘ Type of toxins produced by strain are well defined
4. — Technical grade active ingredient (TGAJ) used in test (see Subdivision M for TOAI
definition)
5 Test fish used was rainbow trout
6. The MPCA dose as recommended in Subdivision M
7. Number of rish/treatment level(s) and controls (10 each)
8. Number of treatment groups used (5)
9. — Type of controls listed, including solvent, carrier, or whether negative or positive
1O. Diet and any antibiotica, vitamins or food additives described
11. Observation period (minimum 30 days)
Result reporting:
12. — LC and/or EC value and the no-observed-effect level reported in ppm
13. Raw mortality data provided
14. — Description of observed toxic effects, including death and any other abnormal signs or
behavior
15.__ Pathological changes as noted by gross necropsy examination described
16. — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a are supplemental and may not be required for every study.
C. 197
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Subdivision M
Guideline Ref. No. 154A-20
December 24, 1989
154A- ) Freshwater Aquatic Invertebrate Test
ACCEPTANCE CRiTERIA
Does ur study meet the Ibilowing a ptance aiteria?
1. Raw data in support of final report are available
2. Identity of test MPCA. genus, species and variety, strain history, identification criteria
3. Type of toxins produced by strain are well defined
4. Technical grade active ingredient (TGAI) used in test (see Subdivision M for TGAI
definition)
5. — Test invertebrate used was _______ (preferrably Daphnia )
6. The MPCA dose as recommended in Subdivision M
7. — Number of invertebtates/treatment level(s) and controls (20 each)
8. Number of treatment groups used (*5)
9. Type of controls listed, including solvent, carrier, or whether negative or positive
10.__ Diet and any antibiotia, vitamins or food additives described
11. — Observation period (*30 days or until 20% mortality observed in controls; three broods for
Daphnia: 14 to 21 days)
Result reporting:
12. — Include LC 50 value in ppm and the no-observed-effect level
13. Raw mortality data provided
14. — Descnption of observed toxic effects, including death, immobilization or other abnormal signs
or behavior
15. — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a * are supplemental and may not be required for every study.
C-198
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SuiI’.bI n \1
Guideline Ref \() 1 iA 2U
D .em er -
154A-21 Estuarine and Marine Axunial Test (Estuartne fish)
ACCEPTANCE CRITERIA
Does your study meet the foLlowing acceptance ciiteria’
— Raw data in support of final report are available
2 — Identity of test MPCA. genus. species and variety, strain history, identification criteria
3 Type of toxins produced by strain are well defined
4 — Technical grade active ingredient (TOAI) used in test (see SubdivisiOn M for TCrAI
definition)
5 — Test fish used was estuarine/marine in origin (silverside or sheepshead minnow preferred
species)
6 — The MPCA dose as recommended in Subdivision M
7 — Number of fish/treatment level(s) and controls (‘10 each)
8 Number of treatment groups used (‘5)
9 — Type of controls listed, including solvent, carrier, or whether negative or positive
10 ‘_ Diet and any antibioties. vitamins or food additives described
ii — Observation period (‘minimum 30 days)
Result reporting.
12 — LC and/or EC, 0 value and the no observed -effect level reported in ppm
13 — Raw mortality data provided
14 Description of observed to c effects. including death and any other abnormal signs or
behavior
15 ‘_ Pathological changes as noted by gross necropsy examination described
16 — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a ° are supplemental and may not be required for every study.
C-199
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Subdi\1 Ion .t
Guideline Scf \‘ .f- i-l
Dccembv r
154A.21 Estuanne and Marine Anurn I T t (Estuanne Shrimp)
ACCEPTANCE CRITERIA
Does your sn y meet the foLlowing aaxpunce aitena?
I Raw data in support of final report are available
2 — Identity of test MPCA genus. species and ‘.arietv. strain history. identification criteria
3 Type of toxins produced by strain are well defined
4 — Technical grade active ingredient (TGAI) used in test (see Subdivision M for TOM
definition)
5 — One of the following organisms was tested White. brown, pink, grass or inysid shrimp
6 — The MPCA dose as recommended in Subdivision M
7 — Number of shrimp/treatment level(s) and controls (10 for static, 20 for flow-through)
8 — Number of treatment groups used (5)
9 — Type of controls listed, including solvent, carrier, or whether negative or positive
[ 0 — Diet and any anubioti , vitamins or food additives described
El — Observation period (min lmum 30 days)
Result reporting
12. — LC and/or EC, 0 value and the no-observed-effect level reported in ppm
13 — Raw mortality data provided
14 Description of observed toxic effects, including death and any other abnormal signs or
behavior
15 _ Pathological changes as noted by gross necropsy examination described
16 Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a are supplemental and may not be required for e iy study.
C-200
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Sub ’-1I”isI fl \l
Guideline Ref ‘ )
DeLember 24
154A-22 NooLargct Plant Studies
ACCEFTANCE CRITERIA
Does your study meet the following aaxpunce criteria’
1 Raw data in support of final report are available
— Identity of test MPCA genus. species and arietv, strain history, identification criteria
3 Type of toxins produced by strain are well defined
4 — Technical grade active ingredient (TGAI) used in test (see Subdivision M for TOM
definition)
5 — The MPCA dose was .. the maximum label application rate
6 — Terrestrial use test species six species of Dicor ledoneae of at least four families and four
species of Monocotvledoneae of at least two families
7 — For herbicidal use, test species that include plants of economic
importance and those necessary for maintanace of ecosystem
8 — For aquatic use, test species should include Selenosinim
capncornutum (freshwater green alga), Lemna £! (duckweed), Skeletonema costatum
(marine diatom), Anabaena flos-aguae (blue-green bacterium), and a freshwater diatom
9 — Negative and positive controls included
10 — Observation period
Result reporting
ii — Description of observed phytotoxic/pathogenic effects and any other abnormal signs
12 — Rationale for deviations from protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a • are supplemental and may not be required for ev ry study.
C-201
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G ci c Re: \ f4A 2:
D eemher 4
154A-23 Montarget Insect Test
ACCEPTANCE CRITERIA
Does your study meet the following acceptance cncria?
I — Raw data in support of final report are available
2 — Identity of test MPCA. genus. species and vanety, strain history. identification criteria
3 ‘ Type of toxins produced by strain are well defined
4 — Technical grade active ingredient (TGAI) used in test (see Subdivision M for TOM
definition)
5 — The MPCA dose as recommended in Subdivision M
6 — Test substance incorporated in the diet
7 ‘_ Diet and any antibiotics, vitamins or food additives described
S — Testing was performed on three species of insect larvae representing at least two of the
following groups:
‘parasitic dipterans predaceous mites
predaceous hemipterans predaceous neuropterans
predaceous coleopterans parasitic hymenopterans
9 — Number of insects/treatment level(s) and controls [ (‘10 each) five replicates
10 — Type of controls listed, including solvent. camer. or whether negative or positi e
11 — Observation penod 30 days or 20% mortality in normal controls
Result reporting:
[ 2. — Include LD value reported in micrograms/insect larva and the no-observed-effect le el
13. — Raw mortality data provided
14 — Description of observed toxic effects, including death and any other abnormal signs or
behavior
15 Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Cnteria marked with a • are supplemental and may not be required for every study.
C.202
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uC’UY bL (l \t
Guidehfle Ret ‘s’) .LA•4
Decerit’er 24 1 q g
154A24 Honey Bee Test
ACCEPTANCE CRITERIA
Does ur study meet the following aarptance aiteria?
— Raw data in support of final report are avai1ab e
— Identity of test MPCA genus. species and anerv. strain history. identification criteria
3 ‘_ Type of toxins produced by strain are well defined
- I — Technical grade active ingredient (TO?. .I) used in test (see SubdivisiOn M for TGAI
definition)
5. — The MPCA dose as recommended in Subdivision M
6 — Test substance incorporated in the diet
7 Diet and any antibioii . vitamins or food addiuves described
8 — Testing was conducted on the honey bee, mellifera
9 — Number of bees/treatment level(s) arid controls [ (‘10 each) five replicate5
10. — Type of controls listed, including solvent. carner. or whether negative or positive
11 — Observation period 30 days or 20% mortality in normal controls
Result reporting.
12. — Include LD value reported in micrograms/bee and the no.observed-effeCt level
13 Raw mortality data provided
14 — Description of observed toxic effects, including death and any other abnormal signs or
behavior
15 — Rationale for deviations from the protocol and the effect. if any, these deviations had on the
ouicome of the study
Criteria marked with a are supplemental and may not be required for e ry study.
C.203
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Suhdi .i Ofl \j
Guideline Ref \o l 4A-25
Dccember 24 1989
154A.25 Terrestrial Wildlife & Aquatic Organism Testing
ACCEPTANCE CRITERIA
Does iu study ni t the following a ptanoe critena?
TIER Ill
— Have submitted or are submitting with this Phase 3 Response the protocol for this study
2 — Have submitted or are submitting with Phase 3 response the results of this study
Criteria marked with a * are supplemental and may not be required for ev ry study.
C-204
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SUb i i L Jfl \1
Guideline Ref \c 154A-26
December i
154A-26 Avian Chronic Pathogewcicy and Reproduction Test
ACCEPTANCE CRITERIA
Does ur study meet the following aeeeptanx criteria’
— Raw data in Support of final report are available
2 Identity of test MPCA. genus, species and variety. strain history, identilication criteria
3 _ Type of toxins produced by strain are well defined
4 — Technical grade active ingredient (TOAI) used in test (see Subdivision M for TGAI
definition)
5 — Test bird used was mallard duck or bobwhite quail
6 — The MPCA dose as recommended in Subdivision M
7 For mallards 2 males and 5 females/pen, replicated by 5 pens/treatment group. or
pens/treatment if individual pairs are used (For bobwhite quail one male and 2 females pen
on individual pairs)
8 — Number of days on which test material given
9 — Type of controls listed, including solvent. carrier, or whether negative or positive
10 No observed effect level determined
11 — Days egg candled for eggshell cracks. embryo viability, and embtyo survival reported
Result reporting.
12. — A summary of reproductive data and reproductive success data for controls and treatment
levels, including 14 day hatchling survivors
13 — Raw mortality data provided
14 — Description of observed toxic effects, including death, weight loss and any other abnormal
signs or behavior
15. — Pathological changes in abnormal birds as noted by gross necropsy examination described
16. — Abnormal organs add lesions cultured for presence of MPCA
17 — Rationale for deviations front the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a • are supplemental and may not be required for cycry study.
C-205
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Sur di’isun \
Guideline Ref \o 15 A.2
December -t l 9
154A-21 Aquatic 1n rtebrate Range T t
ACCEPTANCE CRITERIA
Do your study meet the rouo g aaxptance criteria?
I Have submitted or are submitting with this Phase 3 Response the protocol for this study
2 Have submitted or are submitung with Phase 3 response the results of thiS study
Criteria marked with a e arc supplemental and may not be required for every study.
C-206
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Gutueline Ret NQ I 4A-S
DeL mber ,.i I9 Q
154A-22 Fish Life Cycle Studies
ACCEPTANCE CRITERIA
E ies ur study m t the following aaxpcance aitena?
— Raw data in support of final report are a aiIable
2 — Identity of test MPCA. genus. species and ‘.arietv. strain history. identification criteria
3 Type of toxins produced by strain are well defined
4 — Technical grade active ingredient (TGAJ) used in test see Subdivision M for TOM
definition)
5 — Test fish used was one of the following species
silverside. sheepshead minnow, brook trout. fathead minnow, bluegill sunfish. channel
catfish
6 — The MPCA dose as recommended in Subdivision M
7 — Number of fishitreatmenc level(s) and controls (10 each)
8. — Number of treatment groups used (‘5)
9 — Type of controls listed, including solvent, carrier, or whether negative or positive
10 ‘ Diet and any anubiotica. vitamins or food addici es described
11 — Observation period one complete reproductive cycle
Result reporting:
12 — Weight and condition of all surviving hatchlings
13 — LC 10 andlor EC , value and the no-observed-effect level reported in ppm
14 — Stage of life cycle itt which effects occurred
IS — Description of observed toxic effects, including death and any other abnormal signs or
behavior
16 ‘_ Pathological changes as noted by gross riecropsv examination described
[ 7 — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a • are supplemental and may not be required for every study.
C-207
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Subdt’i ion \1
OuiieIir e Re \ 15.l.A.9
DeLember - I.Q 9
154A-29 Aquatic ELxs tcm Test
ACCEPTANCE CRITERIA
Does your study meet [ be following a ptaooe aiiena 1
— Have submitted or are submming with this Phase 3 Response the protocol for this study
2 — Have submitted or are submitting with Phase 3 response the results of this study
Criteria marked with a are supplemental and may not be required for every study.
C.208
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Guidetifle Re \o 4-\ : i
1 9
154A-31 NontarZet Plant Suidies
ACCEPTANCE CRITERIA
1)xs )vur study meet the foLloiiing auepIan aitena?
I — Have submitted or are submitung with this Phase 3 Response the protocol for this tudv
Have submitted or am submitting with Phase 3 response the results of this study
Criteria marked with a • are suppLemental and may not be required for cveiy study.
C209
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Sur .i ion \
GuideiLne Ref \o 154A-33
December 24 t9 9
154A-33 Sui u1ated and Actual Field Tests (Birds, M mals)
ACCEPTANCE CRITERIA
Do your study m t the !oLlo ng aazptanz cxitena?
TIER IV
— Have submitted or are submitting wtth this Phase 3 Response the protocol for this study
2 — Have submitted or are submitting wuh Phase 3 response the results of thLS study
Criteria marked with a ‘ are supplemental and may noi be required for e c study.
C-210
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.‘ fl ‘ t
Guidet1t C Ret
DcLernt’er 2-1 H ’
154A-34 Simulated and Actual Field Tests (Aquauc Orgaxusms)
ACCEPTANCE CRITERIA
Does ui study tueet the foLlowing aaeptance aiteria?
— Have submitted or are submitting with thiS Phase 3 Response the protocol for this siuth
Have submitted or are submitting with Phase 3 response the results of this studs
Cnteria marked with a • are supplemental and may not be required for every study.
C-211
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urUi. ,r \
Guideline Ref o 4A-35
DeLember 24 l’ S
154A-35 Simulated and Actual cld Tests (Insect Predators, PaI Ites)
ACCEPTANCE CRITERIA
Does your study meet the following a xcpIance criteria?
— Have submitted or are submitting v ith this Phase 3 Response the protocol for this studs
2 — Have submitted or are submitting with Phase 3 response the results of this study
Critena marked with a • are supplemental and may not be required for evei ’ study.
C-212
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Guideline Ref \o I5- A4i5
DeLembe; : yS9
154A-36 S&IDILJated aod Actual Field Tests (1ns Pollinators)
ACCEPTANCE CRITERIA
Does your study me i the following pcan z criteria?
— Have submitted or are submilting ‘ - ith this Phase 3 Response the protocol for this studs
— Have submitted or are submittLng with Phase 3 response the results of this Study
Criteria marked with a • are supplemental and may not be required for every study.
C-2 13
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SuhUi i iOn
Cuidehne Ref \Q l , A-tO lhrou2h 155 .\ 12
Decem er 24 V $9
155A-tO through 155A-12 Enviroamental Expression
ACCEPTANCE CRITER [ A
Does your study t the followuig accepLance criteria’
Since no currently registered microbial pesticides have ethibited significant adverse effects in Tier I
Ecological Effects Studies, no Tier II Environmental Expression Studies have been required n support
ol registration Since techniques (or conducting environmental expression studies are specific to the
unique charactenstics of the microbial pesticide, it is not possible to set forth generic acceptance criteria
at this time. However. all studies should include basic identification data
Product name and trade name (if different)
— EPA Registration Number if registered
3 Taxonomic descnption of the active ingredient
4 — Description of analyttcal method for isoLation, enumeration, and identification of the
microbial pesticide, including a full analysis of the limits of detection of the analytical
method.
Criteria marked with a • are supplemental and may not be required for ev.ery study.
C-214
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Guideline Ret “. ) l ’ LB t )
December ..l 1989
1S1B-lO Product Identity
ACCEflANCE CRITERIA
Does your study meet the following acceptance criteria’
Product name and trade name (if different)
— Name. nominal concentration. and certified limits (upper and lower) for each acti e
ingredient and each intenuonall -added inert ingredient
3 — Name and upper certified limit (or each impurity or each group of impurities present at
0 l c by weight and for certain to cologicatlv significant impurities (e g, microbial toxins.
dioxins. nltrosamines) present at <0.1%
4 — Purpose of each active ingredient and each intentionally-added inert
5 Chemical name from Chemical Abstracts Index of Nomenclature and Chemical Abstracts
Service (CAS) Registry Number for each active ingredient and, if available, for each
intentionally-added inert
6 Product name, trade name, and common name (if established) for each active ingredient
7 — Molecular, structural, and empirical formulas, molecular weight or weight range. and any
company assigned experimental or internal code numbers (or each active ingredient
S — Description of each beginning material in the manufacturing process
— EPA Registration Number if registered; for other beginning materials, the following:
— Name and address of manufacturer or supplier
Brand name, trade name or commercial designation
— Technical specifications or data sheets by which manufacturer or supplier describes
composition, properties or toxicity
9 — Genus and species (and strain, subspecies. isolate. etc., LI applicable) from which the
biochemical was isolated or with which it is commonly associated
10 — Specificity of biochemical activit , the mode of action, and field rates at which the
biochemical is active/proposed (units at/A, etc.)
11 — Similarnv to the naturally-occurring biochemical. if not derived from a biological entity.
12 — Au updated Confidential Statement of Formula must be provided (EPA Form 8570.4 rev.
9i 87)
[ 3 Mv known or suspected hazards of the biochemical to man, the environment, or nontarget
species.
Criteria marked with a * are supplemental and may not be required for every StUdy.
C-215
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urW .Is )r \t
Guideline Ref \o 151 B-i 1
Dt mher -
1518-il Manufacturing Proc s
ACCEPTANCE CRITERLAI,
Does your study meet the following auxplance cntena’
— Descnpuon of manufacturing process or exiractionitsolation steps if obtained from a
biological entity
2._ Statement of whether batch or continuous process. if applicable
3 — Relative amount of beginning materials and order in which they are added
4 — Description of equipment
5 — Description of physical conditions (temperature, pH, pressure, humidity) controlled in each
step and the parameters that are maintained
6 — Statement of whether process involves intended chemical reactions
7 — Flow chart with chemical equations for each intended chemical reaction
8. — Duration of each step of process
9. — Description of purification procedures
10 — Description of measures taken to assure quality of final product including identity of the
biological source, if applicable
11. — A clear presentation of the stage at which inerts are intentionally added, if and when any
concentration is effected, the material to be used as the manufacturing use product (MP),
whether MP registration is sought. and whether a TGAI/MP is sold andlor shipped.
Critena marked with a • are supplemental and may not be required for evqry study.
C-216
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-
GuLdeLine Ret No 1 lB. 12
DcLemher -t iQ 9
151B-12 D cussioa of Formation of Unintended Ingredients
ACCEPTANCE CRITERIA
Does your study m t the fouowing azcpcance criteria?
1 — Discussion of formation of impurities based on established chemical theory addressing (1)
each impurity which may be present at 2 0 1% or was found at 0 1% by product analyses
and (2) certain toxicologically significant impurities present at < 01% by weight
Criteria marked with a • are supplemental and may not be required for every study.
C-217
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Subdi’.isR fl \t
Guideline Ref \t) if 1B-13
December 24 1Y
1518 -13 Analysis of Samples
ACCEPTANCE CRITERIA
Does ur study meet the following ac ptance cnteria?
— Five or more representative samples (batches in case of batch process) analyzed for each
active ingredient and all impurities present at 0.1%
2 — Degree of accountability or closure > 98%
3 — Analyses conducted for certain trace toxic impurities at lower than 0 1% (examples.
nicrosamines in the case of products containing dinitroanhlines or containing secondar or
tertiary amines/alkanolamines plus nitrites, polyhalogenated dibenzodiOXlflS and
dibenzofurans) [ Note that in the case of nitrosammes both fresh and stored samples should
be analyzed.)
4 — Complete and detailed description of each step itt analytical method used to analyze abo e
samples
5 — Statement of precision and accuracy of analytical method used to analyze above samples
6 Identities and quantities (including mean and standard deviation) provided for each analyzed
ingredient
7 — The test material is to be the purest pesticidal grade commeroally produced prior to
intentional addition of inerts. Generally, this test material is the same as that used for
certain nontarget and human hazard testing and is identical to, or equivalent to the technical
grade. Any differences from the test substance used for hazard testing should be noted.
Criteria marked with a • are supplemental and may not be required for eve y study.
C.218
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V
Guideline Ret \o 151B-I
Decemi er -t QSY
151B-15 Cerufi3tioci of Lumis
ACCEYFANCE CRITERIA
Does your study meet the following aa ptance aiteria’
I — Upper and lower certified limits proposed for each active ingredient and intentionally added
inert along with explanation of how the limits were determined
— Upper certified limit proposed for each impurity present at > 0.1% and for certain
toxicologically significant impurities at < 0 1% along with explanation of how each limit is
determined
3 — Analytical methods to verify certified limits of each active ingredient and impurities (latter
not required if exempt from requirement of tolerance or if generally recognized as safe by
FDA) are fully descnbed
4 — Analytical methods to verify certified limits validated as to their precision and accuracy
Criteria marked with a are supplemental and may not be requLred for every study.
C.219
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Suhdi’ .lsiOfl 1
Guidehne Rd o l5lB•
December t
151B-17 Physical and Chemical Properues
ACCEPTANCE CRITERIA
Does your study m t the following a ptance criteria’
A. Color
— Verbal descnption of coloration (or lack of it)
— Any intentional coloration also reported in terms of Munsell color system
B Physical State
— Verbal description of physical state provided using terms such as solid. granular.
volatile liquids
Based on visual inspection at about 20-25°C
C. Odor
— Verbal description of odor (or lack of it) using terms such as garlic-hke.
characteristic of aromatic compounds°
Observed at room temperature
D Melting Point
— Reported in
— Any observed decomposition reported
E. Boing Point
— Reported in °C
— Pressure under which B.?. measured reported
— Any observed decomposition reported
F Density, Bulk Density, Specific Gravity
— Measured at about 20-25°C
— Density/bulk density reported in g.’ml the specific gravity of liquids reported with
reference to water at 20°C [ NOTE: For a solid in particulate form a measurement
of bulk density may be substituted for measurement of densily.j
G. Solubility
— Determined in distilled water, n-octanol and representative polar and non-polar
soMnLs, including those used in formulations and analytical methods for the
p dde
— Measured at about 20-25°C
— Reported in g/lOOinl (other units like ppm acceptable if sparingly soluble)
H. Vapor Pressure
— Measured at =25°C (or calculated by extrapolation from measurements made at
Criteria marked with a • are supplemental and may not be required for every study.
C-flU
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SuN .IL . bI )fl ‘ 1
Guideline Ref \‘ 15 B-l
DeLenther .i l’ 9
higher temperature if pressure too lo to measure at :5’C)
— Expenmental procedure described
Reported in mm Hg (torn or ocher con encional units
I pH
— Measured at about O-5 C
— Measured foflo ing dilution or dispersion in distilled ater
J Stabtliv
— Sensitivity to metal ions and metal determined
Stability at normal and elevated temperatures
— Sensitivity to sunlight determined
K. Flammability
— Flash point reponed in F or °C
flame extension or flame projection reported to nearest centimeter or nearest inch
L. Storage Stability
— Product stored in u.s commercial package or smaller one of same construction and
materials
— Amount of active ingredient determined in product at beginning and end of test
period (duration of at least one year for a product which degrades sufficient
duration to support expiration date)
— Any deterioration or degradation products determined
— Product examined for physical changes at end of test
— Product stored at aoouc 20 2S*C (and 50 relative humidity if permeable packaging)
under warehouse conditions reflecting expected storage
— Report includes duration and conditions of storage, quantitative analyses of active
ingredient. and identification of any deterioration, degradation products. or physical
changes (and consequences of latter on safe handling and use of product)
\4 \iscOSitv
— Determined at about 20-25°C
— Reported in poises, stokes, or other conventional units
N Miscibility
— Determined at about 20-25°C
Product mixed with petroleum solvents whose composition reflects those on label and
at rate on label
— M ture examined for separation after 30 minutes
Criteria marked with a • are supplemental and may not be required for evciy study.
C-221
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SucUi’ i iOfl t
GUIdeline Ref \o l5lBJ
Dec mher 4
0 CurrostOri CharacteriStic
Data on corrosion charactenStI provided (experimental method described !
reasonable explanation given for lack of corrosiveness based on nature of product
(e g, lack of extreme pH. unreacti e)
P 0ctanol 1 watet Partition Coefficient
Measured at about 2O-25 C
— Experimentally determined and description of procedure pvided (preferred method-
45 Fed Regi.ster 77350)
— Data supporting reported value provided
Criteria marked with a • are supplemental and may not be required for evciy study.
C-222
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Subdi isiofl t
Guideline Ref \o 15B-U)
Decembc r - 1’ 9
152B-LO Acute Oral Toxicity
ACCEPTANCE CRITERIA
Does your study m t the following cepUnce critena’
Technical form of the active ingredient tested (for reregistration only)
2 Al least 5 young adult ratsdSeX/grOup
3 — Dosing. single oral dose or in fractions over 24 hours
4 Vehicle control if other than water
5 Doses tested. sufficient to determine a toxicity cata i or a limit dose (5000 mg/kg)
6 — Individual observations at least once a day.
7 — Observation period to last at least 14 days. or until all test animals appear normal whichever
is longer.
8 — Individual daily observations.
9 individual body weights.
10 Gross necropsy on all animals
Criteria marked with a • are supplemental and may not be required for every study.
C-223
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U Ul’.i jr \t
Guideline Ref ‘ o 152B-l 1
December 24. 19S9
152B-lI Acute Dermal Toxcuy
ACCEYTANCE CRITERIA
Do your study meet the following a ptan critena?
Technical form of the acti e ingredient tested (for reregistratton only)
2 ‘__. At least 5 animals/sexigroup
3 ‘.... Rats 200-300 gm. rabbits 20-30 kg or guinea pigs 350-450 gm
4 Dosing, single dermal.
S — Dosing duration at least 24 hours
6 ‘_ Vehicle control. only if toxicity of vehicle is unknown
7 — Doses tested. sufficient to determine a toxicoty cacagory or a limit dose (2000 mg/kg)
S — Application site clipped or shaved at least 24 hours before dosing
9 — Application site at least 10% of body surface area.
10. Application site covered with a porous norurrnaung cover to retain test matenal and to
prevent ingestion
11 — Individual observations at least once a day
12. — Observation penod to last at least 14 days, or until all test animals appear normal whichever
is longer.
13 — Individual daily observatIcr
14 Individual body weights.
15 ‘ Gross necropsy on all animals
Criteria marked with a ‘ are supplemental and may not be required for ever - idy.
C-224
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J’ .i ‘ n
Guideline Re( \ ) 152B4
DeLember ..i i 89
1528-12 Acute Enbalauoci ToEcity
ACCEPTANCE CRITERIA
Dixs your study meet the following acceptance aitena’
Technical form of the active ingredient tested. (for reregistratiofl only)
2. — Product is a gas. a solid which may produce a significant vapor hazard based on toxicir and
expected use or contains particles of inhalable size for man (aerodynamic diameter 15 urn or
less).
3 At least 5 young adult rats/sex/group
4 ‘ Dosing, at least 4 hours by inhalation.
5 Chamber air flow dynamic, at least 10 air changes/hour. at least 19% oxygen content
6 — Chamber temperature. 22° C ( 2°), relative humidity 40-60%.
7 — Monitor rate of air flow.
8 — Monitor actual concentrations of test material in breathing zone.
9 Motor aerodynamic particle size for aerosols.
10 — Doses tested, sufficient to determine a toxicity category or a limit dose (5 mg/I.. actual
concentration of respirable substance).
11 — Individual observations at least once a day
12 — Observation penod to last at least 14 days. or until all test animals appear normal whichever
is longer.
13 — Individual daily observations.
14 Individual body weights.
15 ‘ Gross necropsy on all animals.
Criteria marked with a ‘ are supplemental and may not be required for ev;ry study.
C•225
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SubdL’1s on \
Guideline Ref \o 15B.13
December . Q 9
1523-13 Primary Eye Erntauon
ACCEPTANCE CRiTERiA
Does your study meet the following atxeptan criteria’
— Technical form of the active ingredient tested (for reregistration only)
2 — Study not required if material is corrosive, causes severe dermal imtation or has a pH of 2
or? 115
3 ‘ 6 adult rabbits
4 — Dosing, instillation into the conjunctival sac of one eve per animal.
5 Dose. 0.1 ml if a liquid. 0 1 ml or not more than 100 mg if a solid, paste or particulate
substance.
6 — Solid or granular test material ground to a fine dust.
7 — Eyes not washed for at least 24 hours.
8 — Eyes examined and graded for irritation before dosing and at 1, 24, 48 and 72 hr. then daily
until eyes are normal or 21 days (whichever is shorter).
9 — Individual daily observations
Criteria marked with a are supplemental and may not be required for ev,ry study.
C.226
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L\1l ‘r \t
Guideline Rec \o I 2B-t4
DeL mh r .- l 8
1528-14 Primary Dermal Irritation
ACCEPTANCE CRITERIA
Does your Study meet the following aaeptance critena’
1 — Technical form of the active ingredient tested (for reregistration only)
2 Study not required if material is corrosi e or has a pH of s 2 or 115
3 • 6 adult animals
4 — Dosing. single dermal
5 Dosing duration 4 hours
6 — Application site shaved or clipped at least 24 hour prior to dosing.
7 — Application site approximately 6 cr&
8 — Application site covered with a gauze patch held in place with nonirIltauflg tape
9 — Material removed, washed with water, without trauma to application site
10 — Application site examined and graded for irritation at 1, 24, 48 and 72 hr, then daily until
normal or 14 dat ’s (whichever is shorter).
Li _ Individual observations for the entire day of dosing.
12._ Individual daily observations
Criteria marked with a • are supplemental and may not be required for every study.
C221
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S ’ . mn \j
Guideline Ref \o 15B- 15
Deu niher 24 1989
152.B-15 Uyper cnsitrvity
ACCEFT’ANCE CRITERIA
Does your study m t the following accepIan aitena?
1. Technical form of the active ingredient tested (for reregistration only)
2 — Study not required if material is corrosive or has a pH of < 2 or 11 5
3 — One of the following methods is utilized,
— Freunds complete adjuvant test
— Guinea pig ma omization test
— Split adjuvant technique
Buehler test
— Open epicutaneous test
Mauer optimization test
— Foolpad technique in guinea pig
— Other test accepted by OECD (specify)
4 — Complete description of test
5._ Reference for test.
6. — Test followed essentially as described in reference document.
7 Positive control included
Cntena marked with a • are supplemental and may not be required for every study.
C-228
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Suhdi ision \1
Guideline Ret \c, 15B-1
Decemher 4 lY
152B-17 Genotoxicity
ACCEPTANCE CRITEREA
Does your study meet the blowing aa piance aitena?
General Requirements
— Technical form of the active ingredient tested
2. — Negative, solvent and/or vehicle control(s) for the test system.
3 Positive control(s) for the test system.
4 — Fully identified test system. species. strain, source etc.
5 — Fully described method for maintaining test system.
6 — Fully described method for preparing test environment and administering test compound
7 — Fully described metabolic activation system. if required..
8 — Determination of maximum and range of concentrations/doses used under test conditions
9’ Criteria for determination of a positive effect
Test Specific Requirements
Salmonella reverse mutation assay
1 — Minimum of (our strains, TA98, TAIOO, TA1535 and TA1536 (alternatives need rationale).
2 — Strain specific positive controls.
3 — Highest concentration limited by toxicity, solubility or 5000 ug/plate.
.4 ‘ At least 5 different concentrations of test material at adequate intervals.
5.’_ A single positive response confirmed by testing over a narrow range of concentrations
6.’ At least three plates experimental point.
Criteria marked with a ‘are supplemental and may not be required for every study.
C-229
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Suodi’ i iori \l
Guideline Ref \o 152B-IS
D Lember 4 1q89
152B-18 Immunotonoty
ACCEPTANCE CRITERIA
Does your study meet the following a ept .anee aitena?
Technical form of the acti e ingredient tested
2 AL least 10 yourid adult rat.s or mice (either sex) per dose group per immunological
parameter measured.
3 — Dosing duration daily for at least 30 das.
4 — Doses tested include signs of to city at high dose.
5 Doses tested include a NOEL for immunotoxlcity.
6 ‘_ Analysis for test matenal stability, homogenicuy, and concentratiOn in dosing medium.
7 _ Positive control group wuh a known immunosuppressant if required to venfy assay sensitivity
8. Individual daily observations.
9 Individual body weights
10 Food and water consumption.
ii Hematolo ’
— hematocnt — erythrocyte count
— hemoglobin — total and differential leucocytC count
a..—, platelet count
12 — Spleen weights.
13 — Thvtnus weights.
14 — ainical biochemistry
— glucose
serum glutanc pyruvic Transaminase
‘ urea nitrogen
albumin
— total serum protein
15 — Spleen and thymus cellulanty and cell viability.
16 — Bone marrow cellulanty
17 — Histopatholo ’ of all lesions.
18. — Antibody plaque forming cell assay Immunoglobuhn (IgO, 1gM) quantificaton.
19 — One way mixed lymphocyte reaction assay effect of test substance on induced alluyted type
hypersensitivity reaction cytoto c T.hymprophoryte assay.
20 — Natural killer cell activity.
21 — Mi ophage numbers and function.
Critena marked with a • are supplemental and may not be required for e y study.
C-230
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c’Ui b fl
Guideline Ref “,o 15 :3-19
Decemrer 4 1989
152B-19 Mutagezucny
ACCEFTANCE CRITERIA
Does your study m t the following xeptance altena?
Gene mutation in somatic cells in culture
Highest concentration limited by toxicity (10.20% relative survival), solubilitv or 5000 ug/mI
2 s At least 4 different concentrations of test material to yield a concentration related toxic
effect.
3 — Determination of the number of cell cultures used.
In vitro mammalian cvto2eneti
— Highest concentration limited by toxicity (e g reduced mitotic activity; alteration of cell ccle.
cvtotoxiclty), solubility or 5000 ug/mI.
2 Multiple concenirtions used to define the response.
3 e At least two independent cultures for each experimental point.
5 — Determination of culture harvest time.
In ‘.ivo mammalian cvtogeneti . bone marrow
— At least 5 male and 5 female animals per experimental group.
2. — Highest dose limited by to city or 5000 mgixg.
3 — Determination of sampling times.
Aberrations, a) one treatment . 3 times in range of 6-48 hours after treatment
adequately spaced wuh central sample at 24 hour (may be altered based
on cell cycle time). b) repeated treatments - samples taken 6 and 24
hours after last treatment (may be altered based on cell cycle time .
Micronucleus; Samples taken 3 times, starting not earlier than 12 hours after the last
treatment and at appropriate intervals following the first sample. hut
not beyond 72 hours
4 — Micronucleus assay, at least 1000 polychromattC erythrocytes/animal scored. Ratio of poly to
rtorniochromatic determined by counting 200-1000 er throctes (1000 OECD).
Mv mutagenicity test with suggestive or greater positive results/activity shall be submitted reqardless of
missing essential items.
Criteria marked with a • are supplemental and may not be requited for every study.
C-231
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152B-20 Subchrocuc Feeding
ACCEPTANCE CRITERIA
Does your study met the following aczxptance criteria?
SubdL\i ion ‘.1
Guideline Ret \o 15B-0
De mher -t 1Q89
I Technical form of the acuve ingredient tested
2. Al least 10 rodents or 4 nonrodentslsexlgroup (3 test groups and control group)
3 — Dosing duration daily for 90-days or 5 days/week for 13 weeks
4 — Doses tested include signs of toxicity at high dose but rio lethality in nonrodents or a limit
dose if nontoxic (1000 m kg).
S — Doses tested include a NOEL
6 Analysis for test matenal stability, homogeneity and concentratiOn in dosing medium
7 — Individual daily observations.
8 — Individual body weights
9 Individual or cage food consumption.
10 Opthalmoscopic e minauon (at least pretest and at term) control and high dose.
11 Clinical patholo ’ data of 12 & 13 at termination for rodents, and for non.rodents at the
beginning, then either monthly or midway, and at termination.
12. — Hernatolo ’.
Ervthrocvte count
— Hemoglobin
Hematocnt
13 — Clinical chemistry.
Alkaline phosphatase
— Alanine aminotransferase
— Aspartate aminotransferase
Creatinine kinase
‘ Lactic dehydrogenase
Glucose
Bilirubin
• — Cholesterol
• Creannine
— Leucocyte count
Differential count
— Platelet count (or clotting measure)
Total Protein
Albumin
— Urea nitrogen
— Inorganic phosphate
Calcium
• Potassium
Sodium
* Chloride
14’_ Urinalysis, only when indicated by expected or observed activity. As scheduled in 11.
Blood Total bilirubiri
— Protein * Urobilirubin
— Ketonc bodies — Sediment
— Appearance — Specific gravny (osmolality)
Glucose Volume
15. — Individual necropsy of all animals.
Critena marked with a ‘ are supplemental and may not be required for ev ry study.
C.232
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: Lr ’
152B-21)
4 l9 9
16 — Histopatholo of the follov ing tissues performed on aU nonrodeflls and cm ro&len s iii
control and high dose animals. all animals that died or were killed on study, all gross sionS
on all animals. target organs on all animals and lungs. liver and kidneys on all other anima’s
aorta
eves
caecum
colon
duodenum
— brainf
skin
heart
— testest
— pituitary
ileum
trachea
— gall bladder
— jejunum
bone marrow
— livert
— lung
— lymph nodes
stomach
— mammary gland
— spleent
musculature
— epididymis
— adrenaist
urinary bladder
peripheral nerve
— kidnevst
— esophagus
— ovariest
oviduct
— pancreas
rectum
spinal cord (3x)
thyroid / parathylOlds
— salivary glands
thvmus
— accessory sex organs;UterUS
t organs weighed
Criteria marked with a ‘ are supplemental and may not be required for every study.
Guideline Ref \o
December
C-233
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152B-2 1 Repeated Dermal Dose Toxicity (90-day)
ACCEPTANCE CRITERIA
Does your study meet the following ptance criteria’
Subdi isiOfl \t
GuldeiU 1e Ref .o 152B-N
Decemt er : 98
I — Technical form of the acti e ingredient tested
2 — At least 10 animals/sex/group (3 test groups and control group).
3 — Dosing duration at least 6 hour/day daily for 90 days or S days/week for 13 weeks
4 — Application site at least 10% of body surface area.
5 — Doses tested include signs of toxicity at high dose, no or minimal dermal imlanon. minimal
lethality or a limit dose (I000mg’lg) if nontoxic.
Doses tested include a NOEL
Individual daily observations
Individual body weights.
Individual or cage food consumption
Opthalmoscopic examination (at least pretest and at term) control and high dose
Clinical patholo ’ data of 12 & 13 in all animals at termination.
12 — Hematolo ’.
— Erythrocvte count
— Hemoglobin
Hematocrit
13 — Clinical chemistry.
_ Alkaline phosphatase
— Alanine aminotransferase
— Aspartate aminotransferase
‘ Creatinine kinase
a_ Lactic dehydrogenase
Glucose
Bilirubin
• Cholesterol
$ Creatuune
14.’_ Urinalysis, only when
Blood
Protein
Ketone bodies
— Appearance
Glucose
15. — Individual necropsy of all animals.
Leucocvte count
• Differential count
— Platelet count (or clotting measure)
Total Protein
Albumin
— Urea nitrogen
— Inorganic phosphate
Calcium
* Potassium
Sodium
• Chloride
Criteria marked with a ‘ are supplemental and may not be required for ev ry study.
6
7
8
9.
10
11
indicated by expected or observed activity. As scheduled in ii.
Total bthrubin
• Urobilirubin
Sediment
— Specific gravity (osmolality)
• Volume
C.234
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aorta
eves
caecum
colon
duodenum
— braint
skin
— heartf
— esest
— pituitary
ileum
trachea
— gall bladder
jejunum
bone marrow
— Irvert
lungf
— lymph nodes
stomach
— mammary gland
— spleent
muscula iure
— epididymis
— adrenaist
— urmary bladder
peripheral nerve
kidrtevst
esophagus
ovariest
— oviduct
— pancreas
— rectum
— spinal cord (3x)
thyroid / parathyroids
salivary glands
— thymus
— accessory sex organs;uterus
t organs weighed.
Critena marked with a • are supplemental arid may not be requsred for cvcry study.
Suhdm n \
Guideline Ref \o l B-l
December 4 1989
— l-{tscopatholo r of the following tissues performed on aLl nonrodeflis and roden1s all controt
and high dose animals, all animals that died or were kjUe i on study. alt gross lesions on sit
animals. target organs on all animals and lungs. liver and kidneYs on all other animals
C-235
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“I
Guideline Ref No l B-
Decerni er l - 89
1523-22 Subchronic Lnhalation Tox]city (90-day)
ACCEPTANCE CRITERIA
Does ur study mccl the foLlowing pIaD criteria?
I Technical form of the acti e ingredient tested (for reregistration only)
2 — Product is a gas, a solid which ma produce a significant vapor hazard based on to ]cit and
expected use or contains particles of inhalable size for man (aerodynamic diameter 15 urn or
less).
3 — At least 10 young adult rats/sexigroup
4 — Dosing, 6 hours per day, 5 days per week for 13 weeks.
5 — Food and water should be withheld dunng dosing.
6 °_ Chamber air flow dynamic. at least 10 air changesi1 our. at least 19% oxygen content
7 — Chamber temperature. 220 C (±2°). relative humidity 40-60%.
8 °........ Alternatively, oro-nasal or head only exposures may be used.
9 — Monitor rate of air flow
10 — Monitor actual concentrations of test material in breathing zone.
II. — Monitor aerodynamic particle size for aerosols
12. — individual daily observations.
[ 3 — Individual body weights.
14 — Individual or cage food consumption
15 Opthalmoscopic examination (at least pretest and at term) control and high dose
16 — Clinical patholo ’ data of 17 & 18 in all animals at termination.
[ 7 Hematolo .
— Ervthrocvte count — Leucocvte count
— Hemoglobin ‘ Differential count
— Nematocnt — Platelet count (or clotting measure)
18 — Clinical chemistry.
° Alkaline phosphatase — Total Protein
— Alanine aminotransferase — Albumin
— Aspartate aminotransferase — Urea nitrogen
‘ Creatinine kinase — Inorganic phosphate
‘ Lactic dehydrogenase — Calcium
— Glucose Potassium
— Bilirubin — Sodium
S Cholesterol ‘ Chloride
• Creatinine
Critena marked with a ‘ are supplemental and may not be required for every study.
C-236
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SUC’dI’.bI@fl ‘ .t
Gu det1fle Ref o l52B-
December 4 i9 9
— Unnalsis. only when indicated by expected or obser.ed acti it’ As scheduled in 16
— Blood — Total bilirubin
— Protein Urobilirubin
— Ketorie bodies — Sediment
Appearance Specific gravity (osmolality)
— Glucose ‘ Volume
20 — Individual necropsy of all animals
21 — Histopatholo ’ of the following tissues performed on all nonrodeniS and on rodents all
control and high dose animals. all animals that died or were killed on study, all gross lesions
on all animals. target organs on all animals and lungs. liver and kidneys on all other animals
aorta
eves
caecum
colon
duodenum
— braint
skin
— heartt
— testest
— pituitary
ileum
trachea
— gall bladder
— jejunum
bone marrow
— livert
— lungt
— lymph nodes
— stomach
— mammary gland
— spleent
— musculature
— epididymis
— adrenalst
— urinary bladder
— peripheral nerve
— kidneyst
— esophagus
— ovan j
oviduct
— pancreas
— rectum
— spinal cord (3x)
thyroid I parathyroids
— salivary glands
— thymus
a cessoty sex orgafls.utefllS
f organs weighed.
Criteria marked with a • are supplemental and may not be required for every study.
l
C-237
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Sub .1i’.
Guideline Ref No 15:9-23
DeLember 2- 10S9
1523-23 Teratology
ACCEPTANCE CRITERIA
Does your study m t the foLlowing a piance criteria?
I — Technical form of the active ingredient tested
— At least 20 pregnant animals/dose group for mice. rats or hamsters are available At least 12
pregnant animals/dose group for rabbits are available ( 3 test groups and control group)
3 — At the high dose, overt maternal effects such as slight weight loss are reported as significant
(or a limit dose is given. 1,000 mg,kg).
4 At the low dose, no developmental toxicity is reported.
5 — Dosing duration is at least during the period of major organogenesis. but may extend up to
one day prior to term.
6 Analysis for test material stability, homogeneity and concentration in dosLng medium
7 — Individual daily observations
8. — Individual bodY weights.
9. — Individual food consumption.
LU — Necropsy on all animals
11. — Individual uterine examination including number of fetal deaths, early and late resorptions
and numbers of viable fetuses per sex.
12. — All ovaries examined to determine number of corpora lutea.
13 — Indivtdual litter weights and/or individual fetal weights per sex/litter.
14 — Individual fetus external examination
15 Individual fetus skeletal examination for 113 to 1/2 of each litter for rodents and all for all
rabbits.
16 — Individual fetus soft tissue examination.
Cnzeria marked with a • are supplemental and may not be required for every study.
C .238
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—-“- , V
Guideline Ref \o I 2B-2-
December 4 l9t 9
152B-24 EmmunOtOxlCUy — Tier [ 1
ACCEPTANCE CRITERIA
Checklists are inappropriate at this time because consultation with the Agency is recommended prior to
determining which specific tests are to be performed
Does your study m t thc fotlowi.ng aurptan a teria’
See Special Instructions.
Criteria marked with a • are supplemental and may not be required for every study.
C-239
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Subdi - s ori \1
Gudehne R f “.o 1523 -6
December 2-4 89
1528-26 Chronic Feeding
ACCEPTANCE CRITERIA
Does your study meet the foUowing a xepcance criteria?
Technical form of the active inaredienc tested
2 At least 20 rodents or 4 nonrodents/sexigroup (3 test groups and control group)
3 Dosing duration in rodents minimum 12 month nonfood use. 24 months food use, in
nonrodents minimum 12 months’
4 — Doses tested include signs of toxicity at high dose but no lethality in r iOnrodents or a limit
dose if nontoxic (1,000 mg/kg).
Doses tesled include a NOEL
Analysis for test material stability, homogeneuv and concentrauon in dosing medium
Individual daily observations.
Individual body weights.
Individual or cage food consumption.
Opihalmoscopic exanunation (at least per test and at term) control and high dose.
ainical pathology data (or all nonrodents and at least 10 rodents/group consisting of 12, 13
& 14.
12 — Hematology at 6 month intervals consisting of at least:
Ervthrocvte count — Leucocvte count
— Hemoglobin e Differential count
Hernatocnt Platelet count (or clotting measure)
13 — Clini..al chemistry at 6 month intervals consisting of at least:
• Alkaline phosphatase Total Protein
Alanine arninotransferase
Aspariate aminotransferase
Creannine kinase
Lactic dehydrogenase
Glucose
Bilirubin
Cho l terol
Creannine
14 — Urinalysis at 6 month intervaLs consisting of at least:
— Blood — Total bilirubin
— Protein S..-..— Urobiltrubtn
— Ketone bodies Sediment
— Appearance SpecifIc gravity (osmolahty)
— Glucose * Volume
15. — Individual necropsy of alt animals.
Criteria marked with a • are supplemental and may not be required for evçry study.
6
7
8
9
10 ’
11.
S
a
I
Albumin
Urea
— Inorganic phosphate
Calcium
• Potassium
Sodium
* Chionde
C.240
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Guideline Ret N ; i53-5
December 4 YS9
16 — }-iistopathOlO i of the follo iflg tissues performed on all rionrodefliS and On rodents all
control and high dose animals all animals that died or v ere killed Ofl stud’. all gross lesiocis
on all animals. target organs on all animals arid lungs, li er and kidneys on all other snimals
— peripheral nerve
— kidneyst
— esophagus
ovaries
oviduct
— pancreas
rectum
— spinal cord (3x)
— thyroid / parathyrOldS
— salivary glands
t organs weighed
Six month dog studies may be acceptable.
Cnteria marked with a are supplemental and may not be required for every study.
aorta —
eves
caecum
colon
duodenum
— braint
skin
heart
— testesf
— pituitary
ileum
trachea
— gall bladder
jejunutfl
bone marrow
— livert
— lung
— lymph nodes
stomach
mammary gland
— spleen
— musculature
— epididymis
adrenats
urinary bladder
— thymus
— accessory sex organS;UterUS
C.241
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GuideLine Ref \ i53B-29
D Lember 24 1°89
152B-29 Onoogenictty
ACCEPTANCE CRITERIA
Does your study meet the following aczzpiance criteria’
I — Technical form of the active ingredient tested
2 — At least 50 animals/sex/grOup ( 3 test groups and control group)
3 — Dosing duration is at Least 18 months for mice and 24 months for rats
4 — Number of survivors in any group does not fall below 50% at 15 months for mice, 18
months for rats or 25% at 18 months for mice, 24 months for rats.
5 — Doses tested include an MTD or limit dose if nontoxic (1.000 mgilcg).
6 DOSeS tested include a NOEL
7 ..... ..... .. Analysis for test matenal stability, homogeneity and concentratiOn in dosing medium
8 — Individual daily observations.
9 — Individual body weights.
10 — Individual or cage food consumption.
II. — Individual necropsy of all animaLs.
12. — Blood smear from 10 animals/sex/dose at 12 and 18 months and termination. Differential
count high dose and controls. all other doses if high dose shows patholo ’.
13 — Histopatholo ’ of the following tissues performed on all interim sacrifice animals. all con ol
and high dose animals, all animals that died or were killed on study, all gross lesions on
animals. target organs on all animals and lungs, liver and kidneys on all other animals.
— aorta — jejunum — peripheral nerve
— eves — bone marrow — kidnevsj ’
— caecum — liverf — esophagus
colon lung — ovaries
— duodenum — lymph nodes — oviduct
— braint — stomach — pancreas
— skin — mammary gland — rectum
— heart — spleent — spinal cord (3x)
— testest — musculature — thyroid / parathyroids
— pituitary — epididymis — salivary glands
— ileum adrenals — thymus
— trachea — urinary bladder — accessory sex organs;uterUS
— gall bladder
organs weighed
Cnteria marked with a • are supplemental and may not be required for every study.
C.242
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UiL . i’2fl \
Guideline Ref ‘ o 1 3B- ai
DecemNer . .t 1989
1536-3(a) Chemical Identity
ACCEPTANCE CRITERIA
Does your study meet the following a epunce aitena’
Product name and trade name (if different)
2 — Name, nominal concentration, and certified limits (upper and lower) for each act l%e
ingredient and each intentionally-added inert ingredient
3 — Name and upper certified limit for each impurity or each group of impurities present at
0.1% by weight and for certain toxicologically significant impuntles (e.g., dioxins,
nitrosamines) present at < 0.1%
4 — Purpose of each active ingredient and each intentionally-added inert
5 — Chemical name from Chemical Abstracts Index of Nomenclature and Chemical Abstracts
Service (CAS) Registry Number for each active ingredient and, if available, for each
intentionally-added inert
6. — Product name, trade name, and common name (if established) for each active ingredient
7 — Molecular, structural, and empirical formulas, molecular weight or weight range, and any
company assigned experimental or internal code numbers for each active ingredient
8. — Description of each beginning material in the manufacturing process
— EPA Registration Number if registered for other beginning materials, the following:
— Name and address of manufacturer or supplier
— Brand name, trade name or commercial designation
— Technical specifications or data sheets by which manufacturer or supplier descnbes
composition, properties or toxicity
Criteria marked with a • are supplemental and may not be required for evciy study.
C.243
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Subdr.blon \t
Guideline Ref \o 153B-3ftt
D c mbcr 1989
153B-.3(b) Directions for Use
ACCEPTANCE CRITERIA
Does your study meet the following acceptance criteria’
These criteria are intended to ser’.e as guides for registrants to determine whether adequate directions
for use are available with respect to residue chemistry considerations for each acti’ve ingredient
Registrants should check each line which has been addressed properly by product labels. A written
rauonale. explaining why that particular item is not pertinent, should be provided for each item that has
not been addressed. Since items 6-12 on this list apply to specific types of uses. the registrant may
simply write NA (not applicable) on the line before those types of uses which are not included on any
label.
1. — Each formulation to be used identified with the concentration of active ingredient indicated
(% by weight for solid formulations. pounds active ingredient per gallon for liquids)
2 — All crops which are to be treated with each formulation clearly identified
3. — Tolerance or exemption from tolerance proposed or established for each crop on label
Impractical or unrealistic use restrictions not included (eg., resincting feed use of corn forage
not practical due to high economic value and common practice of feeding to livestock)
5 — Names and quantities of stickers, spreaders, or other adjuvants to be added to spray solution
6 — Field and t rchard crop directions
— Application rate in quantity of formulation and pounds active ingredient per acre
(For row or band treatments indicate if the rate refers to the area treated or to the
entire field)
— Spray volumes to be used per acre
— Maximum number applications per year or growing season
— Minimum interval between successive applications
— Minimum interval between last application and harvest (preharvest interval= PHI)
— For orchard crops additional information for full coverage sprays (quantity of
formulation and pounds active ingredient per 100 gallons spray) and concentrated
sprays (amount active per acre should be related to tree size)
7 — Aerial, ultra low volume (ULV) and mist spray directions include spray concentration.
amount of active ingredient per acre, and spray volume per acre
8 — Postharvest fumigation directions
— Dosage expressed in weight fumigant per volume of storage space or weight fumigant
per unit weight of commodity treated
— Temperature. pressure and duralion of exposure specified
— Geomcuy and airtighiness of containers described
— Aeration procedures and time of aeration specified
— Minimum interval between successive applications
Criteria marked with a are supplemental and may not be required for evejy study.
C-244
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Lt fl \1
Ciuidel fle Ref \o 153B-3b1
Decemt er 4 l9 9
9 — Animal treatment directions
— Concentration of pesticide in treatment solution
— Quantity of spray, pour-on solutiOn. dust. etc to be applied per animal
— Amount of time animal to be held in dip tank
— Frequency of treatment
— Maximum number of treatments
— Preslaughter interval (interval between last treatment and slaughter) specified
(interval longer than 3 days usually not considered practiCal)
10 — Aquatic use directions
— Dosage expressed in quantity of formulation and pounds active ingredient per surface
acre or parts per million pesticide in the water (in latter case the amount product
per surface area should be related to average pond depth)
Detailed description provided for specialized equipment
— Minimum distance specified from treated area to potable water or imgatlon intake
pipe
If oxygen depletion problems. proportion of pond to be treated and required interval
between treatments
— Maximum number of applications per year
— Minimum interval between applications
ii — Food handling establishment use directions
— Type of establishment that may be treated
Dilution instruCtionS and spray concentration
— Type of application equipment and mode of application (space spray, directed spray
to crevices, spot treatment. etc)
— Dosage limitations including cubic and square foot limitations
— Frequency of treatment
— Time of treatment (eg.. after-hours in restaurants)
— lnstruCtiOflS concerning removal and covering of food, dishes and utensils
— Cleanup procedures before food processing. preparation or serving resumes
12. — Agncultural premises treatments
— Description of areas to be treated (eg.. feed lot, milking room. animal barn)
— Dosage specified (pounds per unit volume treated for fogging applications;
concentration of active ingredient in spray solutions; weight active ingredient per unit
area of feed Lot)
— Frequency of treatment
— Directions as to whether animals should be removed during treatment
Criteria marked with a • are supplemental and may not be required for cvçiy study.
C.245
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Suodi’ ’)fl \t
Guideline Ref \o 53B-3(C)
Detem er 4 i 9
153B-3(C) Mature of the Residue-Plants
ACCEPTANCE CRITERIA
Does your study meet the following acceptance
— Pesticide radiolabeled in non-labile portion of molecule (tritium label strongly discouraged)
2 — SpecifIc activity sufficient to permit detection of low residue levels (001-01 ppm range)
3 — Radiochemically pure grade of active ingredient
4 — Pesticide applied to plant in manner simulating expected use
5 Total radioactivity measured in plant parts used for human food or animal feed
6 — Extractability of residue into solvents (preferably those used in analytiCal enforcement
method) determined
7 Most of radioactivity extracted or exhaustive attempts (acid. base, enzyme) made to do so
8 Major components or portion of the terminal residue identified (preferably by at least two
techniques-e.g. TLC, HPLC, MS)
Criteria marked wtth a * are supplemental and may not be required (or evçry study.
C-246
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SupU ’- fl \t
GuldeiLne Ref \o 153B -3(di
December 4 1q89
1538-3(d) Nature of the Residue-Animais
ACCEflANCE CRITERIA
Does your study meet the following acceptance cntena’
— Pesticide radiolabeled in non-labile portion of molecule (tritlum label stronglY discouraged’)
2. — Specific activity sufficient to permit detection of low residue levels (001-0 1 ppm range
3. — Radiochemically pure grade of active ingredient used for dosing
4 — Pesticide administered orally to animals for at least three consecutive days (or applied
externally according to label directions for dermal use)
5 — Animals not preconditioned by dosing with unlabeled material
6 — Animals sacnficed within 24 hours of final dose or application
7 — Total radioactivity measured in edible tissues (muscle, fat. liver, ruminant kidney) and milk
or eggs
8. — Extractability of residue into solvents (preferably those used in analytical enforcement
method) determined
9. — Most of radioactivity extracted or exhaustive attempts (acid, base, enzyme) made to do so
10. — Major components or portion of the terminal residue identified (preferably by at least two
techniques-e-g, TLC, KPLC, MS) in edible tissues and milk or eggs
Criteria marked with a • are supplemental and may not be required for eveiy study
C-247
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Suhdi ision \.l
Guideline Ref \ l 3B.3ie and f
Decemt er 4
153B-3(e and I) Residue nalytical Method
ACCEPTANCE CRITERIA
Does OUf study meet the fouowing a eptance criteria’
I List of equipment. reagents and standards provided along with U S sources/suppliers of same
2 — Instrumentation and operating conditions described
3 — Detailed descnpnon of each sep in procedure to enable use by competent analyst unfamiliar
with method
4 — Discrete response for analyte
5 — Control values reasonably low compared to tolerance
6 — Adequate recoveries (generally 70%) obtained for fortifications at the tolerance level
7 — Recoveries don’t vary significantly from sample to sample
8. — Evidence that aged residues extracted by procedure (may reference work in metabolism
studies)
9. — Data showing that method releases and recovers bound residues (if latter of toxicological
concern)
10 — Requirements for regulatory methods (as opposed to those used only for collecting residue
data)
— Does not require untreated commodity as blank
— Does not require internal or procedural sandard to correct for recoveries (Addition
of internal standard in final step just prior to injection is acceptable for calibration
of retention times. However, use of internal standard throughout entire procedure
to correct for recoveries is not acceptable unless data are available on numerous
samples of each matrix to show analvie and internal standard behave identically in
each step.)
Does not require exotic equipment or reagents
— Reasonably rapid in execution
— Specific to measure residue in presence of other reasonably expected pesticides
— Sensitive in relation to tolerance
Confinnaroiy method available
— Method not claimed to be Confidential Business Information
— Does not use hazardous reagents (justification needed for use of benzene as solvent
or diazomethane as inethylacmg agent)
Criteria marked with a ‘ are supplemental and may not be required for every study.
C.248
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Subdivision M
Guideline Ref. No. 153B-3(g)
December 24, 1989
153B-3(g) Storage Stability
ACCEPTANCE CRITERIA
Does ur study m t the following a xeptance aiteria?
1 — Sample preparation and fortification described (or in cases where samples with weathered
residues are used, history of crop and pesticide treatment provided) -
2. — Storage conditions specified (temperature, containers, form of r.a.c., lighting, etc.)
3 — Dates of fortification (or harvest in case of field treated samples), placement into storage,
sampling, and residue analysis provided
4. All components of total toxic residue fortified into commodity (or present from field
treatment with pesticide) and measured by validated analytical method(s) (description of
latter provided or referenced)
Criteria marked with a are supplemental and may not be required for every study.
C-249
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Subdivision M
Guideline Ref. No. 153B.3(h)
December 24, 1989
153B-3(h) Magnitude of the Residue - Potable Water
ACCEPTANCE CRiTERIA
Does your study meet the following a ptancc aiteria?
1. — Bodies of water treated according to label directions leading to maximum residues:
— Maximum application rate and number of applications
— Minimum retreatment intervals
2. — Data reflect modes of application on label: e.g., aerial, specialized equipment, treatment of
pond margins only.
3 — Representative formulations examined
4 — Water sampled in center of treated area and at various points to show effect of distance on
residue level
5. — Water sampled on day of treatment and later periods to show effect of time on residue level
6. Sufficient number of trials conducted to determine an appropriate tolerance or maximum
residue level (factors to consider include “geographic representation”, seasonal and yearly
variations, sizes and types of bodies of water treated, the toxicity of the pesticide)
7 — Total toxic residue measured in water by validated analytical method (description provided or
referenced)
8. — Description of sample treatment, collection, handling and storage provided (covering periods
from treatment to sampling and from sampling through residue analysis)
9. — Dates of treatment, sample collection, entry into storage, and residue analysis
10. — Storage stability data available reflecting storage of water samples (may be referenced to
separate study)
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-250
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Subdivision M
Guideline Ref. No. 153B-3(i)
December 24, 1989
153B-3(i) Magnitude of the Residue - Fish
ACCEPTANCE CRITERIA
Does your study meet the ftfflowing a ptanee criteria?
1. — Fish maintained in water containing maximum pesticide residues permitted by product labels
2. — Fish sampled on day of treatment and later periods to show effect of time on residue level
and the time required for residues to plateau
3. — Residue data obtained for bottom feeders (e.g., catfish) and predators (e.g., bass) in the case
of fish and for molluses (e.g., clams, oysters) and crustaceans (e.g., shrimp, crabs) in the case
of shellfish
4. — Total toxic residue measured in fish by validated analytical method (description provided or
referenced)
5. — Analyses reflect fish commodity definitions in the Pesticide Analytical Manual, Volume I
6. — If pesticide used in estuanne areas, data provided for whole fish protein concentrate and
smoked, canned or other processed fish products
7. — Description of pesticidal treatment of water and collection, handling and storage of fish
samples provided
8. — Dates of treatment, sample collection, entry into storage, and residue analysis
9. Storage stability data available reflecting storage of fish samples between sample collection
(i.e., sacnfice) and residue analysis (may be referenced to separate study)
Criteria marked with a * are supplemental and may not be required for every study.
0.251
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Subdivision M
Guideline Ref. No. 153B.3(j)
December 24, 1989
1538 -3 (j) MagnItude of the Residue - Irrigated Crops
ACCEPTANCE CRITERIA
Does your study meet the following a ptanee criteria?
1. — Crops irrigated with waler containing maximum pesticide residues permitted by product labels
2. Representative crops from each crop grouping in 40 CFR 180.34(1) examined for residues
resulting from treated irrigation water
3. Crops sampled for residues within one day of irrigation
4. Ar least two field trials conducted for each representative crop
5. — Total toxic residue measured in crops by validated analytical method (description provided or
referenced)
6. Description of pesticidal treatment of irrigation water, irrigation technique, and collection,
handling and storage of crop samples provided
7. Dates of pesticidal treatment of water, irrigation of crops, harvest of crops, entry of crop
samples into storage, and residue analysis
8 — Storage stability data available reflecting storage of crop samples (may be referenced to
separate study)
Criteria marked with a * are supplemental and may not be required for every study.
C-252
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Subdivision M
Guideline Ref. No. 153B.3(k)
December 24, 1989
153B-3(k) Magnitude of the Residue - Food Handling
ACCEPTANCE CRITERIA
Does your study meet the following a eptanee aiteria?
1. — Pesticide applied in manner leading to highest residues in food that could result from actual
use (factors to consider include spray concentration; type of spray [ space, spot, crack and
crevice]; interval between sprays if multiple applications permitted; food covered or
uncovered)
2. — Foods representative of those to be present in treated facilities analyzed for pesticide
residues (e.g., in case of cafeteria include meat, baked goods, oily food [ butter, cheese], fresh
fruits and vegetables)
3. — Foods sampled for residue analysis as soon as permitted after application and at various later
times
4 — Pesticide residue measured in representative foods by validated analytical method (description
provided or referenced)
5. — Description of sample treatment, collection, handling, and storage conditions/duration
provided
6. Storage stability data available reflecting storage of food samples from collection until residue
analysis (may be referenced to separate study)
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-253
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Subdivision M
Guideline Ref. No. 153B-3(l)
December 24, 1989
153B -3(I) Magnitude of the Residue - Meat, Milk, Poultry, Eggs
ACCEFFANCE CRITERIA
Does your study meet the following a ptance criteria?
1. Compound(s) fed corresponds to residues expected on feed items
2. Several dosages examined with one approximating intake expected from treated feed items,
one or more representing exaggerated intake, and one control group
3. — Multiple animals included in each dosage group (preferably at least 3 for cattle and 10 for
poultry)
4. — Duration of dosing adequate to ensure residues plateau in milk or eggs (preferably at least 4
weeks)
5. — Animals sacrificed within 24 hours of final dose
6. — Total toxic residue measured in edible tissues (muscle, fat, liver, cattle kidney) and milk or
eggs of both control and treated animals using validated analytical method (description of
latter given or referenced)
7. — Analytical method sufficiently sensitive (0.01-0.05 ppm)
8. — Descnption of handling and dosing of animals, feed consumption, and sample collection,
handling and storage provided
9. — Storage stability data available reflecting storage of tissue, milk and egg samples prior to
residue analysis (may be referenced to separate study)
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-254
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Subdivision M
Guideline Ref. No. 153B-3(m)
December 24, 1989
153B-3(m) Reduction of Residues
ACCEFI’ANCE CRITERIA
Does your study meet the following aa ptanee aiteria?
1. — Description of sample history provided for the raw agricultural commodity (r.a.c.). This
could include items such as the pesticidal treatment (application rate, preharvest interval,
where grown, etc.), date and location of purchase, and storage conditions from harvest or
purchase until processing/washing/cooking.
2. Detailed descnption of residue reduction procedure used on r.a.c. Such procedures may
include trimming, washing, peeling, cooking, or storage of the commodity under simulated
commercial or home food preparation conditions.
3. — Description of handling and storage of samples from time of residue reduction procedure
until analysis for residues
4 — Total toxic residue measured with a validated analytical method (descnption provided or
referenced) in the r.a.c. both before and after the residue reduction procedure
5. — Storage stability data available reflecting storage of r.a.c. and aprocessedR samples from time
of residue reduction procedure until analysis for residues
Criteria marked with a * are supplemental and may not be required for every study.
C-255
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Subdivision M
Guideline Ref. No. 153B -3(n)
December 24, 1989
153B-3(n) Magnitude of the Residue - Crop Field Trials
ACCEPTANCE CRiTERIA
Does ur study m t the following aa ptanoe aiteria?
1. Crop treated according to label directions leading to maximum residues:
— Maximum application rate and number of applications
Minimum retreatment intervals
— Minimum preharvest interval
— Minimum spray volume
2. — Data reflects modes of application on label: ground, aerial, oil diluents, use of adjuvanis such
as surfactants and spreader/stickers
3. — Representative formulations examined
4. — Trial locations represent all principal growing regions (adequate “geographic representation”)
OR minor crop for which regional registration is accepted
5. Sufficient number of trials conducted to determine an appropriate tolerance or maximum
residue level (factors to consider include seasonal and yearly variations, differences in
varieties and cultural practices, the importance of the crop, the toxicity of the pesticide and
its ability to translocate, and the type of use-e.g., early seasonlpre -emergence versus late
season foliar)
6 — Total toxic residue measured by validated analytical method (description provided or
referenced) on all parts of crop used for food or feed (except for feed items which are
restricted by product labels)
7. — Description of sample treatment, collection, handling (including any washing or trimming)
and storage provided (covering periods from planting to harvest and from harvest through
analysis)
8. Dates of treatment, harvest, entiy into storage, and residue analysis
9 — Storage stability data available reflecting storage of crop samples (may be referenced to
separate study)
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-256
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Subdivision M
Guideline Ref. No. 153B-3(o)
December 24, 1989
153B -3(o) Magnitude of the Residue - Proccssed Food
ACCEPTANCE CRiTERIA
Does your study meet the following a p1ance criteria?
1. — The raw agricultural commodity (r.a C.) samples that were processed contained field-treated
detectable residues (preferably at or above the tolerance) Q r.a.c. was treated in the field
at exaggerated rates in an attempt to get detectable residues Q spiked r.a.c.’s were used
with data available to show that r.a.c. residues exist solely on the surface
2. — The r a.c. was processed using procedures simulating commercial practices
3. The total toxic residue was measured with a validated analytical method (description provided
or referenced) in the r.a.c. and all its byproducts listed in Table 2 of the Residue Chemistry
Guidelines
4 — Descnption of pesticidal treatment and processing of r.a.c. and collection, handling and
storage of samples provided
5. — Storage stability data available reflecting storage of r.a.c. and byproduct samples (may be
referenced to separate study)
Criteria marked with a * are supplemental and may not be required for every study.
C-257
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Subdivision M
Guideline Ref. No. 154B-6
December 24, 1989
1548-6 Acute Avian Oral Test
ACCEPTANCE CRITERIA
Does your study meet the following ac ptance aiteria?
TIER I
1. — Raw data in support of final report are available
2. — Chemical name of test substance
3. — Technical grade active ingredient (TGAI) used in test
4. — Percent active ingredient
5. — Name and percent of related compounds and impurities
6. — The pesticide dose as recommended in Subdivision M
7. — Test bird used was mallard duck or bobwhite quail
8. — Number of birds/treatment level(s) and controls (10 each)
9. — Type of controls listed, including solvent, carrier, or whether negative or positive
10. — Diet and any antibiotica, vitamins or food additives described
11. — Observation period (>14 days)
Result reporting:
12. — Include LD value in mg/kg and the no-observed-effect level
13. — Raw mortality data provided
14 — Description of observed toxic effects, including death and any other abnormal signs or
behavior
15. — Pathological changes as noted by gross necropsy examination described
16 Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a * are supplemental and may not be required for every study.
C-258
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Subdivision M
Guideline Ref. No. 1543-7
December 24, 1989
1548-7 Avian Dietaiy Test
ACCEPTANCE CR1ThRIA
Does your study meet the following acceptan aiteria?
1 — Raw data in support of final report are available
2. — Chemical name of test substance
3 Technical grade active ingredient (TOAI) used in test
4. — Percent active ingredient
5. — Name and percent of related compounds and impurities
6. — Test bird used was mallard duck or bobwhite quail
7. — Age at the beginning of the test is 5 to 10 days for mallard ducks and 10 to 14 days for
bobwhite quail
8. — The pesticide dose as recommended in Subdivision M
9. — Number of days on test material given (5)
10. Analysis of the pesticide concentration in the diet was performed
11. — Number of birds/treatment level(s) and controls (10 each)
12. — Type of controls listed, including solvent, carrier, or whether negative or posftive
13. — Observation period (>8 days)
Result reporting:
14. — Include LD value in mg/kg and the no-observed-effect level
15 Raw mortality data provided
16 — Description of observed toxic effects, including death, weight loss and any other abnormal
signs or behavior
17. — Pathological changes in abnormal birds as noted by gross necropsy examination described
18 — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-259
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Subdivision M
Guideline Ref. No. 154B-8
December 24, 1989
154B-8 Freshwater Fish LC Test
ACCEPTANCE CRITERIA
Does your study meet the following aeeeptance aiteria?
1. — Raw data in support of final report are available
2. — Chemical name of test substance
3. — Technical grade active ingredient (TGAI) used in test
4. — Percent active ingredient
5. — Name and percent of related compounds and impurities
6. — Test fish used was rainbow trout
7. — Number of fish/treatment level(s) and controls (10 each)
8 — S treatment groups used
9 — The pesticide dose as recommended in Subdivision M
10 — Type of controls listed, including solvent, carrier, or whether negative or positive
11. Observation period (minimum 96 hours)
Result reporting:
12. — LC and/or EC value and the no-observed-effect level reported in ppm
13 — Raw mortality data provided
14. Description of observed toxic effects, including death and any other abnormal signs or
behavior
15 — Pathological changes as noted by gross necropsy examination descnbed
16. — Rationale for deviatioas from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a * are supplemental and may not be required for every study.
C-260
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Subdivision M
Guideline Ref. No. 154B-9
December 24, 1989
154B-9 Freshwater Aquatic Invertebrate LC,, Test
ACCEPTANCE CRITERIA
Does your study mccl the following act ptan aiteria?
1, — Raw data in support of final report are available
2. — Chemical name of test substance
3. Technical grade active ingredient (TGAI) used in test
4. — Percent active ingredient
5. — Name and percent of related compounds and impurities
6. Test invertebrate used was _________ (preferrably Daphnia )
7. Pesticide dose as recommended in Subdivision M
8 — Number of invertebtates/treatment level(s) and controls (20 each)
9. 5 treatment groups used
10. — Type of controls listed, including solvent, carrier, or whether negative or positive
11. — Diet and any antibioti , vitamins or food additives described
12. Observation period (4Shr)
Result reporting:
13 — Include LCç 0 value in ppm and the no-observed-effect level
14 — Raw mortality data provided
15. — Description of observed toxic effects, including death. immobilization or other abnormal signs
or behavior
16. — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a * are supplemental and may not be required for every study.
C-261
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Subdivision M
Guideline Ref. No. 154B-1O
December 24, 1989
154B-1O Nontarget Plant Studies (Seed GcrminationlSeedling Emergence)
ACCEPTANCE CRITERIA
Does your study meet the following a ptance aiteria?
I. — Raw data in support of final report are available
2. — Chemical name of test substance
3. — Technical grade active ingredient (TGAJ) used in test
4. — Percent active ingredient
5. — Name and percent of related compounds and impurities
6. — Plants tested were six dictyledoneae species from at least four families and four
monocotyledoneac species from at least two families (corn, soybean, and a root crop must be
included)
7. — Dose rates in ppm and lb al/a at the maximum label rate
8. l0 seeds planted/container
9. — 3 treatment groups used
10 — Type of controls listed, including solvent, carrier, or whether negative or positive
11 — Test duration and observation periods were 5 days for seed germination and two weeks for
seedling emergence
Result reporting:
12. — Percent germination and percent emergence recorded
13. — Description of observed phytotoxic effects
14. — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a * are supplemental and may not be required for every study.
C.262
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Subdivision M
Guideline Ref. No. 154B-10
December 24, 1989
154B-1O Noniarget Plant Studies (Vegetative Vigor)
ACCEPTANCE CRITERIA
Does ur study meet the following acceptance aiteria?
1. — Raw data in support of final report are available
2. — Chemical name of test substance
3. — Technical grade active ingredient (TGAI) used in test
4. — Percent active ingredient
5. — Name and percent of related compounds and impurities
6. — Plants tested were six dictyledoneae species from at least four families and four
monocotyledoneae species from at least two families (corn, soybean, and a root crop must be
included)
7. — Dose rates in ppm and pounds of active ingredient per acre (lb aila) at the maximum label
rate
8. — 10 seeds planted/container
9. — 3 treatment groups used
10. — Type of controls listed, including solvent, carrier, or whether negative or positive
11. — Test duration and observation periods were 2 weeks
Result reporting:
12. — Growth measurements recorded (height and dry weight)
13. — Description of observed phytotoxic effects
14 — Percent effect as compared to control
15. Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a are supplemental and may not be required for every study.
C-263
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Subdivision M
Guideline Ref. No. 154B-11
December 24, 1989
154B-1 Nontarget Insect Testing
ACCEPTANCE CRiTERIA
Does your study mect the killowing a ptanoe criteria?
1. — Raw data in support of final report are available
2. — Chemical name of test substance
3. Technical grade active ingredient (TGAI) used in test
4. Percent active ingredient
5. Name and percent of related compounds and impurities
6. The pesticide dose as recommended in Subdivision M
7. — Contract toxicity study, or:
8. Test substance incorporated in the diet
9. Diet and any antibioties, vitamins or food additives descnbed
10. Testing was performed on three species of insect/larvae representing at least two of the
following groups:
parasitic dipterans, predaceous hemipterans, predaceous coleopterans, predaceous
mites, predaceous neuropterans, parasitic hymenopterans
11. Number of insects/treatment level(s) and controls (10 each) five replicates
12. Type of controls listed, including solvent, carrier, or whether negative or positive
13. Observation penod 48 hr
Result reporting:
14. — Include LD 50 value reported in micrograms/insect larva and the no-observed-effect level
15. — Raw mortality data provided
16. — Description of observed toxic effects, including death and any other abnormal signs or
behavior
17. — Rationale for deviations from the protocol and the effect, if any, these deviations had on the
outcome of the study
Criteria marked with a • are supplemental and may not be required for every study.
C-264
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Subdivision M
Guideline Ref. No. 154B-12
December 24, 1989
154B-12 Terrestrial Wildlife Testing
ACCEPTANCE CRITERIA
Does your study meet the following a xeptanee criteria?
TIER HI
1. — Have submitted or are submitting with this Phase 3 Response the protocol for this study.
2. — Have submitted or are submitting with Phase 3 response the results of this study.
Criteria marked with a are supplemental and may not be required for every study.
C-265
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Subdivision M
Guideline Ref. No. 154B-13
December 24, 1989
154B-L3 Aquatic Anim2I T ung
ACCEPTANCE CRITERIA
Does your study meet the Ibilowing a ptance aiteria?
1. — Have submitted or are submitting with this Phase 3 Response the protocol for this study.
2. — Have submitted or are submitting with Phase 3 response the results of this study.
Criteria marked with a are supplemental and may not be required for every study.
C-266
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Subdivision M
Guideline Ref. No. 154B-14
December 24, 1989
154B-14 Nontarget Plant Studies
ACCEPTANCE CRITERIA
Does vur study meet the following aeeeptance criteria?
1. — Have submitted or are submitting with this Phase 3 Response the protocol for this study.
2. — Have submitted or are submitting with Phase 3 response the results of this study.
Criteria marked with a * are supplemental and may not be required for every study.
C-267
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Subdivision M
Guideline Ref. No. 154B-15
December 24, 1989
154B-15 Nontarget Insect Testing
ACCEPTANCE CRflERLA
Does ur study meet the following a pIanee aiteria?
1. Have submitted or are submitting with this Phase 3 Response the protocol for this study.
2. — Have submitted or are submitting with Phase 3 response the results of this study.
Criteria marked with a * are supplemental and may not be required for every study.
C-268
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Subdivision M
Guideline Ref. No. 1553-4
December 24, 1989
1553-4 Volatility Study (Lab)
ACCEPTANCE CRITERIA
Does your study meet the following aeeeptanee aiteria?
1 *_ A greenhouse or field volatility study was provided as a substitute for this data requirement.
2. A typical End Use Product (EP) was used, or an adequate explanation provided to justify the
alternative chosen.
3. — Radiopurity and specific activity of Active Ingredient (AI)(if radiolabelled).
4. — The positions of the Al radiotabeling were appropriate (if radiolabelled).
5. — Experiments were conducted with each respectively labelled ring (for compounds containing
more than one ring structure).
6. — Soils were completely characterized, using the USDA classification system.
7 — Foreign soils (if any) were adequately compared with domestic (USA) soils.
8. — Test substance was added to the soil at the highest recommended label rate for a single
application.
9. Sampling intervals were adequate.
10. The decline of the A! was clearly established.
ii. — Material balance was reasonable (>90% - <110%)
12. — Trapping efficiencies were reasonable.
13. — Reported experimental temperature was held constant (±1°C) between 18 and 30°C.
14.*_ VolatLltty was reported in the correct untts (ug!cm 2 ilir).
15. — The concentration of the A! in air was measured.
16 ‘_ The concentration of Al in air was reported in the correct units (nglm 3 or ugim ).
17 _ The vapor pressure was reported in the correct units (TorT, or mm Hg), including the
temperature at which it was determined.
18. — Appropnaie analytical methods were provided.
19.*_ Detection limits were reported.
20. Raw data in support of the Final Report are available.
Criteria marked with a • are supplemental and may not be required for eveiy study.
C-269
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Subdivision M
Guideline Ref. No. 155B-4
December 24, 1989
1558-4 Volatility Study (Field)
ACCEPTANCE CRfl R1A
Does your study meet the Ibilowing a ptanee aiteria?
1. The study was conducted domestically (USA).
2. — A typical End Use Product (EP) was used, or an adequate explanation provided to justi1 r the
alternative chosen.
3. — Test substance was added to the soil at the highest recommended label rate for a single
application.
4. — The test site (including soil type) was typical of actual use; soil was adequately charactenzed
using the USDA classification system.
5. — The site used for this study was clearly shown to have no previous use history involving this
(or closely related) compound (Al).
6. — Environmental conditions were typical, and adequately described.
7. — Sampling intervals were adequate.
8. — The decline of the Al was clearly established.
9 *_ Volatility was reported in the correct units (uglcmZ/llr).
10 — The concentration of the A! in air was measured.
11 _ The concentration of Al in air was reported in the correct units (ng/m 3 or ug/m 3 ).
12.*_ The vapor pressure was reported in the correct units (Torr, or mm Hg), including the
temperature at which it was determined.
13. — Appropnate analytical methods were provided.
14.’_ Detection limits were reported.
15. — A storage stability study was conducted using either spiked field or spiked laboratory samples
to determine the stability of samples under typical lab storage conditions.
Criteria marked with a * are supplemental and may not be required for every study.
C-270
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Subdivision M
Guideline Ref. No. 155B-5
December 24, 1989
155B-5 & 155B-6 Leaching!Msorption/Desorptjon
ACCEPTANCE CRITERIA
Does your study meet the following a ptanee aiteria?
1. — Testing method was Batch Equilibrium or Soil Column.
2. — Analyte was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabelled
(PAIRA).
3. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabelled).
4 — The positions of the Al radiolabeling were appropriate (if radiolabelled).
5. — Experiments were conducted with each respectively labelled ring (for compounds containing
more than one ring structure.
6. — At least 4 soils were used, which had pH’s between 4 and 8.
7. — One soil had a %OM < 1%.
8. — One of the soils was the same type of soil used in the aerobic metabolism study.
9. — Soils were completely characterized using the USDA classification system.
10. — Soils were not sieved using a screen smaller than 2mm.
11. ”_ Soil moisture was maintained at 75% of 1/3 bar during the aging process.
12. — Foreign soils (if any) were adequately compared with domestic (USA) soils.
13. — Test substance was added to the soil at the highest recommended label rate for a single
application.
14’ Matenal balance was reasonable (>90% <110%).
15. — Mobility of parent and all major degradates identified in the laboratory metabolism studies
were determined during each column leaching study.
16 — Raw data in support of the Final Report are available.
For SOIL TLC . (no longer recommended)
1. Reference standards for degradates identified in the laboratory metabolism studies were used
2. R values were reported for parent and degradates.
For SOIL COLUMN...
1. Elution volume was equivalent to 20” (50.8 cm times the cross-sectional area of the column).
2. — Column length was adequate.
3. — Distribution (identity and quantity) of major degradates/metabolites within the column
(including eluale) was determined.
For AGED LEACHING_
1. — Test substance was aged for 30 days or one half-life (whichever is shorter).
2. — A reasonable attempt made to identify the parent and degradates.
Criteria marked with a are supplemental and may not be required for every study.
C-271
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Subdivision M
Guideline Ref. No. 155B-6
December 24, 1989
For BATCH RQUILffiRIUM...
1. — Test substance was equilibrated at a minimum of 4 concentrations, with adequate time
allowed for complete equilibration.
2. — O.OIN Ca solution was used.
3. — Appropriate analytical methods were provided
4. — Detection limits were reported.
5. — K and were reported.
Criteria marked with a * are supplemental and may not be required for every study.
C-272
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Subdivision M
Guideline Ref. No.155B-8
December 24, 1989
1553-8 Ultraviolet Absorption Study
ACCEPTANCE CRITERIA
Does your study m t the following a ptanoe criteria?
1. — Analyte was the Technical Grade (TGAJ) or a Pure Active Ingredient Radiolabelled
(PAIRA).
2. — Radiopunty and specific activity of Active Ingredient (Al) (if radiolabelled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabelled).
4 — Experiments were conducted with each respectively labeled ring (for compounds containing
more than one nng structure and if radiolabeled).
5 — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
6. — If natural sunlight used, record specifics regarding time of year, meteorological conditions
during exposure, etc.
7. — If artiflacial light source used, compare light source to natural light including nature,
intensity, spectrum and length of exposure
8. — Wavelengths <290 nm were filtered out (artificial light source).
9 — Dark controls maintained at same temperature as expenmental chamber.
10. — Volatiles (if any) were trapped.
11 — Sampling intervals were adequate.
12 — Material balance was reasonable (>90% - <110%)
13. — A reasonable attempt was made to identify all photoproducts which exceeded 10% (yield) of
the initial concentration of the Al.
14. — Appropriate analytical methods were provided.
l5. _. Detection limits were reported.
16. — Half-lives were calculated.
Criteria marked with a * are supplemental and may not be required for every study.
C-273
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Subdivision M
Guideline Ref. No.155B-9
December 24, 1989
155B-9 Hydrolysis Study
ACCEPTANCE CRiTERIA
Does your study meet the following aa’eptanoe criteria?
1. — Analyte was the Technical Grade (TGAJ) or a Pure Active Ingredient Radiolabelled
(PAIRA).
2 — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabelled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabdlled).
4. — Solubility of the Al at each pH tested.
5. Concentration of Al (in solution) within the known solubility range of the Al and below 250
ppm
6. — Study was conducted in darkness.
7. — Temperature held at 25±1°C.
8 — Sterility of glassware, reagents, chemicals, etc. was assured.
9. — Volatiles (if any) were trapped.
10. — Solutions were buffered.
11. — Cosolverit (if any) did not exceed 1%.
12. pH of each test solution was reported and was correct (nominally 5,7 and 9) and constant.
13 Sampling intervals were adequate.
14 *_ Study was conducted for one half-life or 30 days.
15. A reasonable attempt was made to identify all degradates/hydrolysates which exceeded 10%
(yield) of the initial concentration of the Al.
16. — Material balance was reasonable (>90% -<110%)
17 — Appropriate analytical methods provided.
18,*_ Detection limits were reported.
19 — Half-lives were calculated.
Critena marked with a S are supplemental and may not be required for every study.
C-274
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Subdivision M
Guideline Ref. No. 155B-10
December 24, 1989
155B-1O Aerobic Soil Metabolism Study
ACCEPTANCE CRITERIA
Does your study meet the following a ptance a iteña?
1. — Analyte was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabelled
(PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabelled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabelled).
4. — Experiments were conducted with each respectively labeled ring (for compounds containing
more than one ring structure and if radiolabeled).
S. — Soil moisture was maintained at 75% of 1/3 bar during the aging process.
6. — Foreign soils (if any) were adequately compared with domestic (USA) soils.
7. Soil was a sandy loam, silt loam, or representative of intended use area.
8 — Soils were completely characterized, using the USDA classification system.
9. — Soils were not sieved using a screen smaller than 2mm.
10. Reported experimental temperature was held between 18 and 30°C (±1°C.
11 Study was conducted in darkness.
12. Sampling intervals were adequate.
13. — Material balance was reasonable (>90% - <110%)
14 — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
15. — Study was conducted until patterns of decline of parent and patterns of formation/decline of
degradates were established or for no more than one year.
16 *_ Detection limits were reported.
17 — A reasonable attempt was made -- perhaps with multiple solvent systems •- to extract
metabolites/degradates).
18. — A reasonable attempt was made to identify the parent and all degradates [ which exceeded
10% (yield) of the initial concentration of the Al or 0.01 ppm (whichever is smaller)
19. — Half-life of parent was calculated.
20. Appropriate analytical methods were provided.
Criteria marked with a ‘ are supplemental and may not be required for evety study.
C-275
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Subdivision M
Guideline Ref. No. 155B-11
December 24, 1989
155B-11 Aerobic Aquatic Metabolism Study
ACCEPTANCE CRfl’ERIA
Ekes vur study m t the following aecepsan aiteria?
1. — Analyte was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabelled
(PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (AI)(if radiolabelled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabelled).
4. Experiments were conducted with each respectively labelled ring (for compounds containing
more than one ring structure).
5* Water was adequately characterized.
6. — Foreign soils/sediment (if any) were adequately compared with domestic (USA)
soils/sediments.
7 — Sediment was representative of the intended use site.
8. — Both aerobic and aquatic conditions were assured and maintained.
9. — Application rate was consistent with the sensitivityof the analytical method, so that
degradates could be identified.
10. — Reported experimental temperature was held constant (±1°C) between 18 and 30°C).
11. — Sampling intervals were adequate over the 30 days of the study.
12. — A reasonable attempt was made to identif , the parent and all degradates fwhich exceeded
10% of application or 0.01 ppm (whichever is smaller).
13. — A reasonable attempt was made -- perhaps with multiple solvent systems -- to extract
metabolites/degradates.
14 — Half-life of parent was calculated.
15. — Patterns of formation and decline of major degradates were reported.
16. — Material balance was reasonable (>90% - <110%)
17.*_ Detection limits were reported.
18. Appropriate analytical methods were provided.
Criteria marked with a * are supplemental and may not be required for every study.
C-276
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Subdivision M
Guideline Ref. No. 155B-12
December 24, 1989
155B-12 Photolysis in Soil Study
ACCEPTANCE CRITERIA
Does your study meet the following acceptance aiteria?
1 — Analyte was the Technical Grade (TGAJ) or a Pure Activelngredient Radiolabelled (PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabelled).
3. The positions of the Al radiolabeling were appropriate (if radiolabelled).
4. — Experiments were conducted with each respectively labeled ring (for compounds containing
more than one ring structure and if radiolabeled).
5. — Soils were completely characterized, using the USDA classification system.
6. — Soils were not sieved using a screen smaller than 2 mm.
7 *_ Soil moisture was maintained at 75% of 1/3 bar during the aging process.
8. — Foreign soils (if any) were adequately compared with domestic (USA) soils.
9 — At least one of the soils was the same type as was used in the aerobic metabolism study.
10. — Soil was vital (not stenlized).
11. — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
12. — Volatiles (if any) were trapped.
13. — If natural sunlight used, redord specifics regarding time of year, meteorological conditions
during exposure, etc.
14. — If artificial light source used, compare light source to natural light including nature, intensity,
spectrum and length of exposure.
15 — Wavelengths <290 nm were filtered out (artificial light source).
16. — Dark controls maintained at same temperature as exposed soils.
17. Reported experimental temperature was held between 20 and 30°C.
18. — Sampling intervals were adequate.
19. — Material balance was reasonable (>90 - <110%).
20. — A reasonable attempt was made at each sample time to identify all photoproducts which
exceeded 10% (yield) of the initial concentration of the Al.
21 — Appropnate analytical methods were provided.
22. _ Detection limits were reported.
23. — Half-lives were calculated and reported.
Criteria marked with a • are supplemental and may not be required for every study.
C-277
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Subdivision M
Guideline Ref. No. 155B-13
December 24, 1989
155B-13 Photolysis in Water Study
ACCEPTANCE CRITERIA
Does your study meet the fbllowing a ptanee aiteria?
1. — Analyte was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabelled
(PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabelled).
3. — The positions of the Al radiolabeling were appropnate (if radiolabelled).
4. — Solubility of the A!.
5. — Concentration of the AL (in solution) within the known solubility range of the A!, and below
250 ppm.
6. — Volatiles (if any) were trapped.
7. Solutions buffered at the pH of minimal hydrolysis, a pH between 5 and 9.
8. — Cosolvent (if any) did not exceed 1% and was definitely not known photosensitizer.
9. If natural sunlight used, record specifics regarding time of year, meteorological conditions
during exposure, etc.
10. — If artificial light source used, compare light source to natural light including nature, intensity,
spectrum and length of exposure.
11 — Wavelengths <290 nm were filtered out (artificial lightsource).
12 — Dark controls maintained at same temperature as exposed solutions.
13. — Reported expenmental temperature was held at 25±1°C.
14. — Sterility of glassware, reagents, chemicals, etc. was assured.
15. — Sampling intervals were adequate.
16 — Material balance was reasonable (>90% - <110%)
17 — A reasonable attempt was made at all ampling times to identi ’ all degradates/hydrolysates
which exceed 10% (yield) of the initial concentration of the Al.
18. — Appropriate analytical methods were provided.
19 Detection limits were reported.
20. — Half-lives were calculated.
Criteria marked with a * are supplemental and may not be required for every study.
C-278
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SUBDIVISION N
161-1 Hydrolysis . .
161-2 Photodegradation in Water . 284
161-3 Photodegradation on Soil 288
161-4 Photodegradation in Air 292
162-1 Aerobic Soil Metabolism 295
162.2 Anaerobic Soil Metabolism 299
162.3 Anaerobic Aquatic Metabolism 303
162-4 Aerobic Aquatic Metabolism 307
163-1 Leaching/Adsorption/Desorption 311
163-2 Laboratory Volatility 316
163-3 Field Volatility 320
164-1 Terrestnal Field Dissipation 323
164-2 Aquatic Field Dissipation 327
164.3 Forest Field Dissipation 331
164-5 Long-Term Soil Dissipation 335
165.1 Confined Rotational Crops 339
165-2 Field Rotational Crops 342
165.3 Accumulation in Irrigated Crops 346
165-4 Bioaccumulation in Fish 350
165-5 Bioaccumulation in Aquatic Non-Target Organisms . . 354
C .279
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Subdivision N
Guideline Ref. No. 161-1
December 24, 1989
161-1 Hydrol s
ACCEPTANCE CRITERIA
Do your study meet the following aaxptance afteria?
1. Test substance was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2. Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabeled).
4 — Solubility of the Al at each pH tested.
5. Concentration of Al (in solution) within the known solubility range of the Al and below
250 ppm
6. — Study was conducted in darkness.
7. — Temperature held at 25±1°C.
8. — Sterility of glassware, reagents, chemicals, etc. was assured.
9. Volatiles (if any) were trapped.
10 Solutions were buffered.
11. — Cosolvent (if any) did not exceed 1%.
12. pH of each test solution was reported and was correct (nominally 5,7 and 9) and constant.
13. — Sampling intervals were adequate.
14.* Study was conducted for one half-life or 30 days.
15. — A reasonable attempt was made to identify all degradates/liydrolysates which exceeded 10%
(yield) of the initial concentration of the Al.
16. Material balance was reasonable (>90% -<110%).
17 — Appropriate analytical methods provided.
18.*_ Detection limits were reported.
Criteria marked with a * are supplemental and may not be required for every study.
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Guideline Ref. No. 161-1
December 24, 1989
19 — Half-lives were calculated.
Criteria marked with a * are supplemental and may not be required for every study.
C-281
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Subdivision N
Guideline Ref. No. 161-1
December 24, 1989
161-1 Hydrolysis
GUIDANCE FOR SUMMARIZING STUDIES
Items in summ iry should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Conclusions
B. Test Material
1. The chemical structure: (for radiolabeled substances include ‘ ‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3 Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package
C Test Methodology
1. Level of fortification
2. Light conditions during study
3. Temperature range during experiment
4 Sterility measures during experiment (if used)
5 Description of water source and characteristies (including pH, conductivity, hardness, etc)
6 Description of pH’s and buffer solutions
7. Description of cosolvent and concentration (if any)
8. Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
9. Description of volatility trap and analysis procedures (if for radiolabeled substances included)
10. Description of extraction method for parent material and individual degradates
11. Description of extraction efficiency as percent of applied individually for parent and degradates
12. Description of analytical method including standards used and percent recovery
13. Description of storage conditions
C-282
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14 Description of storage stability and results
E. Reported Results
1. Description of matenal balance at sampling intervals as percent of applied
2. Descnption of half-life calculation for parent and degradates
3. Description or graph of formation and decline of parent and degradates
F. Discussion of Results
C.283
Subdivision N
Guideline Ref. No 161.1
December 24, 1989
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Subdivision N
Guideline Ref. No. 161.2
December 24, 1989
161-2 Photodegradation in Water
ACcEJ’IANCE CRITERIA
Does your study meet the following a eptanee criteria?
1. — Test substance was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabeled).
4 — Solubility of the Al.
5. — Concentration of the Al (in solution) within the known solubility range of the Al, and below
250 ppm.
6. Volatiles (if any) were trapped.
7. — Solutions buffered at the pH of minimal hydrolysis, a pH between 5 and 9.
8. — Cosolveni (if any) did not exceed 1% and was definitely not known photosensitizer.
9 — If natural sunlight used, record specifics regarding time of year, meteorological conditions
during exposure, etc.
10 11 artificiaL light source used, compare light source to natural light including nature, intensity,
spectrum and length of exposure.
11. Wavelengths <290 nm were filtered out (artificial light source).
12. — Dark controls maintained at same temperature as exposed solutions.
13 — Reported experimental temperature was held at 25±1°C.
14. — Sterility of glassware, reagents, chemicals, etc. was assured.
15. Sampling intervals were adequate.
16. Material balance was reasonable (>90% - <110%)
17. — A reasonable attempt was made at all sampling times to identil all degradatesibydrolysates
which exceed 10% (yield) of the initial concentration of the Al.
Critena marked with a are supplemental and may not be required for every study.
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Guideline Ref. No. 161-2
December 24, 1989
18. — Appropnate analytical methods were provided.
19._ Detection limits were reported.
20. — Half-lives were calculated.
Criteria marked with a * are supplemental and may not be required for every study.
C-285
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Subdivision N
Guideline Ref. No. 161-2
December 24, 1989
161-2 Photodegradation in Water
GUIDANCE FOR SUMMARIZING STUDIES
items in summary should indude the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Conclusions
B Test Material
1 The chemical structure: (for radiolabeled substances include ‘‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include rathopurity and specific activity)
3. Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package
C. Test Methodolo
I Level of fortification
2. Description of light source
a If natural, describe time of year, meteorological conditions, actual photoperiod and other
variables afi Ling light intensity (include latitude and longitude)
b if artificial, indicate light source, intensity, spectrum, dark/light photoperiod as compared to
natural sunlight and whether or not wavelengths <290 nm filtered out
3 Temperature range during experiment
4. Description of dark controls
5. Description of water source and characteristics (including pH, conductivity, hardness, etc)
6 Description of pH’s and buffer solutions
7. Description of cosolvent and concentration (if any)
8. Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
9, Description of volatility trap and analysis procedures (if used)
10. Description of extraction method for parent material and individual degradates
C-286
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Guideline Ref. No. 161-2
December 24, 1989
11. Description of extraction efficiency as percent of applied individually for parent and degradates
12. Description of analytical method including standards used and percent recovery
13. Description of storage conditions
14. Descnption of storage stability and results
E. Reported Results
1 Description of material balance at sampling intervals as percent of applied
2. Description of half-life calculation for parent and degradates
3. Description or graph of formation and decline of parent and degradates
F. Discussion of Results
C-2S7
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Subdivision N
Guideline Ref. No. 161-3
December 24, 1989
161-3 Photodegradation on Soil
ACCEPTANCE CRITERIA
Does your study meet the following a ptanee aiteria?
1. — Test substance was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2. — Radiopunty and specific activity of Active Ingredient (Al) (if radiolabeled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabeled).
4. — Experiments were conducted with each respectively labeled ring (for compounds containing
more than one ring structure and if radiolabeled).
5. — Soils were completely characterized, using the USDA classification system.
6 Soils were not sieved using a screen smaller than 2 mm.
7 _ Soil moisture was maintained at 75% of 1i3 bar during the aging process.
8. Foreign soils (if any) were adequately compared with domestic (USA) soils.
9. — At least one of the soils was the same type as was used in the aerobic metabolism study.
10 Soil was vital (not sterilized).
11. — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
12. — Volatiles (if any) were trapped.
13. — If natural sunlight used, record specifies regarding time of year, meteorological conditions
during exposure, etc.
14. — Artificial light.
— if artificial light source used, compare light source to natural light including nature,
intensity, spectrum and length of exposure.
If artificial light source used, light/dark photoperiod approximated natural conditions
(12 hr: 12 hi, 16 hr: 8 hr, etc.).
15. — Wavelengths <290 nm were filtered out (artificial light source).
16. — Dark controls maintained at same temperature as exposed soils.
Criteria marked with a * are supplemental and may not be required for every study.
C-288
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Subdivision N
Guideline Ref. No. 161-3
December 24, 1989
17. Reported experimental temperature was held constant (± 1°C) between 18 and 30°C.
18. Sampling intervals were adequate.
19. — Matenal balance was reasonable (>90 - <110%).
20 — A reasonable attempt was made at each sample time to identify all photoproducis which
exceeded 10% (yield) of the initial concentration of the Al.
21 Appropriate analytical methods were provided.
22 . Detection limits were reported.
23. Half-lives were calculated and reported.
Criteria marked with a * are supplemental and may not be required for every study.
C-2S9
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Subdivision N
Guideline Ref. No. 161-3
December 24, 1989
161-3 Photodegradation on Soil
GUIDANCE FOR SUMMARIZING STIJDEES
Items in summaiy should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Conclusious
B. Test Material
I. The chemical structure: (for radiolabeled substances include ‘‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3. Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package
C. Soil description
1. Description of test soil source (including country, state of origin, etc) for each soil used
2 Description test soil type (USDA classification)
3 Description of soil characteristics
a. % sand, silt, clay
b. % organic material
c. pH
d. Cation exchange capacity (meqIlOO grams)
4. Comparison of foreign soil (if used), including soil characteristics (noted above) and microbial
activity, to U.S. soils
5. Description of soil preparation including sieve size and sterilization technique (if used)
6. IndLcate other studies supporting this registration which use this soil type
C-290
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Subdivision N
Guideline Ref. No. 161-3
December 24, 1989
D. Test Methodology
1. Level of fortification
2. Description of light source
a. If natural, descnbe time of year, meteorological conditions, actual photopenod and other variables
affecting light intensity (include latitude and longitude)
b. If artificial, indicate light source, intensity, spectrum, dark/’ight photoperiod as compared to
natural sunlight and whether or not wavelengths <290 nm filtered Out
3 Temperature range dunng expenment
4. Description of dark controls
5. Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
6. Description of volatility trap and analysis procedures (if used)
7. Description of extraction method for parent material and individual degradates
8. Description of extraction efficiency as percent of applied individually for parent and degradates
9. Description of analytical method including standards used and percent recovery
10. Description of storage conditions
11. Description of storage stability and results
E. Reported Results
1 Description of material balance at sampling intervals as percent of applied
2. Description of half-life calculation for parent and degradates
3. Description or graph of formation and decline of parent and degradates
F. Discussion of R ulis
C-291
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Subdivision N
Guideline Ref. No. 161-4
December 24, 1989
161-4 Photodegradation in Air
ACCEPTANCE CRITERIA
Does your study meet the following a ptancc aiteria?
1. — Test substance was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeled).
3. The positions of the AL radiolabeling were appropnate (if radiolabeled).
4. — Experiments were conducted with each respectively labeled ring (for compounds containing
more than one ring structure and if radiolabeled).
5 — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
6 If natural sunlight used, record specifics regarding time of year, meteorological conditions
dunng exposure, etc.
7. — If artificial light source used, compare light source to natural light including nature, intensity,
spectrum and length of exposure
8. — Wavelengths <290 nm were filtered out (artificial light source).
9 — Dark controls maintained at same temperature as experimental chamber.
10. Volatiles (if any) were trapped.
11. Sampling intervals were adequate.
12. — Material balance was reasonable (>90% - <110%)
13. — A reasonable attempt was made to identify all photoproducts which exceeded 10% (yield) of
the initial concentranon of the Al.
14. — Appropriate analytical methods were provided which clearly distinguished between vapor-
phase and solution (if any) phase photoproducts
15. _ Detection limits were reported.
16. — Half-lives were calculated.
Criteria marked with a * are supplemental arid may not be required for every study.
C-292
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Subdivision N
Guideline Ref. No. 161-4
December 24, 1989
161-4 Photodegradation in Air
GUIDANCE FOR SUMMARIZING SThDIES
Items in summary should indude the items discussed in Chapter 2 of this package and the specific items
Listed below.
A. Summary of Condusious
B. Test Material
1. The chemical structure: (for radiolabeled substances include ‘*‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3. Solubility of the test material in water (include temperature and pH)
4 Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package
C. Test Methodology
1. Level of fortification
2. Description of light source
a. If natural, describe time of year, meteorological conditions, actual photoperiod and other variables
affecting light intensity (include latitude and longitude)
b. If artificial, indicate light source, intensity, spectrum, darkIlight photoperiod as compared to
natural sunlight and whether or not wavelengths <290 nm filtered out
3. Temperature range during experiment
4. Description of dark controls
5 Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
6 Description of volatility trap and analysis procedures (if used)
7. a. Description of extraction method for parent material and individual degradates
b. Descnption of method used to distinguish between vapor phase and solution phase degradates.
8 Description of extraction efficiency as percent of applied individually for parent and degradates
9. Description of analytical method including standards used and percent recovery
C-293
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10 Descnption of storage conditions
11. Descnption of storage stability and results
E. Reported Results
1. Description of material balance at sampling interval as percent of applied
2. Description of half-life calculation for parent and degradates
3. Description or graph of formation and decline of parent and degradates
F. Discussion of Results
C-294
Subdivision N
Guideline Ref. No. 161-4
December 24, 1989
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Subdivision N
Guideline Ref. No. 162.1
December 24, 1989
162-1 Aerobic Soil Metabolism
ACCEPTANCE CRITERIA
Does ur study meet the following at ptanee iteria?
1. — Test substance was the Technical Grade (TGAI) or a Pure Active Ingredieni Radiolabeled
(PAIRA).
2 Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeled).
3 — The positions of the A! radiolabeling were appropnate (if radiolabeled).
4. — Experiments were conducted with each respectively labeled ring (for compounds containing
more than one nng structure and if radiolabeled).
5. — Soil moisture was maintained at 75% of 113 bar during the aging process.
6. — Foreign soils (if any) were adequately compared with domestic (USA) soils.
7 — Soil was a sandy loam, silt loam, or representative of intended use area
8 — Soils were completely characterized, using the USDA classification system.
9 — Soils were not sieved using a screen smaller than 2mm.
10 Reported experimental temperature was held constant (± 1°C) between 18 and 30°C.
11 — Study was conducted in darkness.
12 — Sampling intervals were adequate.
13. — Material balance was reasonable (>90% . <110%)
14 — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
15. — Study was conducted until patterns of decline of parent and patterns of formation/decline of
degradates were established or for no more than one year.
16.s_ Detection limits were reported.
Criteria marked with a * are supplemental and may not be required for every study.
C-295
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Subdivision N
Guideline Ref. No. 162-1
December 24, 1989
17 A reasonable attempt was made -- perhaps with multiple solvent systems -- to extract
metabolites/degradates).
18. — A reasonable attempt was made to identi ’ the parent and all degradates [ which exceeded
10% (yield) of the initial concentration of the Al or 0.01 ppm (whichever is smaller)J.
19. Half-life of parent was calculated.
20. Appropriate analytical methods were provided.
Criteria marked with a * are supplemental and may not be required for every study.
C-296
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Subdivision N
Guideline Ref. No. 162.1
December 24, 1989
162-1 Aerobic Soil Metabolism
GUIDANCE FOR SUMMARIZING ST1JD ES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Conclusions
B. Test Material
1 The chemical structure: (for radiolabeled substances include ‘*‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3. Solubility of the test matenal in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package
C. Soil description
I. Description of test soil source (including country, state of origin, etc) for each soil used
2. Description test soil type (USDA classification)
3 Descripuon of soil characteristics
a. % sand, silt, clay
b. % organic material
c. pH
d. Cation exchange capacity (meq/lOO grams)
4. Comparison of foreign soil (if used), including soil characteristics (noted above) and microbial
activity, to U.S. soils
5. Description of soil preparation including sieve size
6. Indicate other studies supporting this registration which use this soil
C-297
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Subdivision N
Guideline Ref. No. 162.1
December 24, 1989
D. Test Methodolo ,
1. Level of fortification
2. Description of treatment technique
3. Description of soil moisture content throughout study
4. Temperature range during experiment
5 Description of method used to initiate and maintain aerobic conditions
6. Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
7. Description of volatility trap and analysis procedures (if used)
8. Description of extraction method for parent matenal and individual degradates
9 Description of extraction efficiency as percent of applied individually for parent and degradates
10 Description of analytical method including standards used and percent recovery
11. Description of storage conditions
12. Description of storage stability and results
E. Reported Results
1. Description of material balance at sampling intervals as percent of applied
2. Description of half-life calculation for parent and degradates
3 Description or graph of formation and decline of parent and degradates
4. Description of aerobicity measurement (Eh) and results if measured.
F. Discussion of Results
C-298
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Subdivision N
Guideline Ref. No. 162-2
December 24, 1989
162-2 Anaerobic Soil Metabolism
ACCEPTANCE CRIThRIA
Do your study meet the following aa ptanee aiteria?
1. — Test substance was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeled).
3. — The positions of the A! radiolabeling were appropriate (if radiolabeled).
4 Experiments were conducted with each respectively labelled ring (for compound containing
more than one ring structure).
Soil moisture was maintained at 75% of 1/3 bar during the aging process.
6 — Foreign soils (if any) were adequately compared with domestic (USA) soils.
7 Soil was a sandy loam, silt loam, or representative of intended use area.
8 — Soils were completely characterized, using the USDA classification system.
9 — Soils were not sieved using a screen smaller than 2mm.
10 — Reported experimental temperature was held constant (± 1°C) between 18 and 30°C
11 — Study was conducted in darkness.
12. — Sampling intervals were adequate.
13. — Material balance was reasonable (>90% - <110%)
14. — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
15 — Treated soil was aged for one half-life or 30 days (whichever is shorter) prior to initiation of
anaerobic conditions.
16. — Study was conducted until patterns of decline of parent and patterns of formation/decline of
degradates were established or for no more than 60 days.
17 *_ Detection limits were reported.
Criteria marked with a * are supplemental and may not be required for every study.
C-299
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Subdivision N
Guideline Ref. No. 162.2
December 24, 1989
18. A reasonable attempt was made to identif r the parent and all degradates [ which exceeded
10% of initial application or 0.01 ppm (whichever is smaller)J.
19. — The unextractable fraction was reasonable (i.e. a reasonable attempt was made -- perhaps
with multiple solvent systems -- to extract metabolites/degradates).
20. — Half-life of parent was provided.
21. — Appropriate analytical methods were provided.
Criteria marked with a * are supplemental and may not be required for every study.
C-300
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Subdivision N
Guideline Ref. No. 162-2
December 24, 1989
162-2 Anaerobic Soil Metabolism
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Condusions
B. Test Material
1. The chemical structure: (for radiolabeled substances include ‘ ‘ to indicate label position)
2 The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3 Solubility of the test matenal in water (include temperature and pH)
4. Reference additional studies that are presented to satis1 ’ this data requirement, that use alternate
radiolabeling and that are included in this summary package
C Soil description
1. Description of test soil source (including country, state of origin, etc) for each soil used
2. Description test soil type (USDA classification)
3. Description of soil characteristics
a. % sand, silt, clay
b. % organic matenal
c. pH
-d. Cation exchange capacity (meq/100 grams)
4. Comparison of foreign soil (if used), including soil charactenstics (noted above) and microbial
activity, to U.S. soils
5. Descnption of soil preparation including sieve size
6 Indicate other studies supporting this registration which use this soil
C-301
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Subdivision N
Guideline Ref. No. 162-2
December 24, 1989
D. Test Methodolo ,
1. Level of fortification
2. Description of treatment technique
3. Description of soil moisture content throughout study
4. Temperature range during experiment
5. Description of method used to initiate and maintain anaerobic conditions
6. Description of sampling intervals including time zero concentration. (indicate hr, day, weeks)
7. Description of volatility trap and analysis procedures (if used)
8. Description of extraction method for parent material and individual degradates
9 Description of extraction efficiency as percent of applied individually for parent and degradates
10. Description of analytical method including standards used and percent recovery
11. Description of storage conditions
12. Description of storage stability and results
E. Reported Results
1. Description of material balance at sampling intervals as percent of applied
2. Description of half-life estimation for parent
3. Description or graph of formation and decline of parent and degradates
4. Description of anaerobicity measurement (Eh) and results if measured.
F. Discussion of Results
C-302
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Subdivision N
Guideline Ref. No. 162-3
December 24, 1989
162-3 Anaerobic Aquatic Metabolism
ACCEPTANCE CRITERIA
Does your study meet the following a eptance criteria?
1. Test substance was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2 Radiopunty and specific activity of Active Ingredient (AL) (if radiolabeled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabeled).
4 — Experiments were conducted with each respectively labelled ring (for compounds containing
more than one nng structure).
5. Water, sediment or soil were completely characterized, using the USDA classification system.
6. — Foreign soils or sediment (if any) were adequately compared with domestic (USA)
soils/sediments.
7. — The maximum length of the study was one year.
8 — Anaerobic conditions were assured and maintained.
9 — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
10. — Reported experimental temperature was held constant (± 1°C) between 18 and 30° C.
ii — Sampling intervals for water and sediment were adequate.
12. A reasonable attempt was made to identify the parent and all degradates [ which exceeded
10% of application or 0.01 ppm (whichever is smaller)].
13. — The unextractable fraction was reasonable (i e. a reasonable attempt was made -. perhaps
with multiple solvent systems -- to extract metabolites/degradates).
14. Half-life of parent was calculated.
15. Patterns of formation and decline of major degradates were reported.
16. — Material balance was reasonable (>90% - <110%).
Criteria marked with a * are supplemental and may not be required for every study.
C-303
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Subdivision N
Guideline Ref. No 162.3
December 24, 1989
17.* Detection limits were reported.
18. — Appropriate analytical methods were provided.
Criteria marked with a * are supplemental and may not be required for every study.
C-304
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Subdivision N
Guideline Ref. No 162-3
December 24, 1989
162-3 Anaerobic Aquatic Metabolism
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Condusious
B Test Material
1. The chemical structure: (for radiolabeled substances include ‘ ‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3. Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this date requirement, that use alternate
radiolabeling and that are included in this summary package
C. Test media
1 Description of each test soil or sediment source (including country, state of origin, etc) for each soil
used
2. Description test soil or sediment type (USDA classification)
3 Description of soil or sediment characteristics
a. % sand, silt, clay
b % organic material
cpH
d. Cation exchange capacity (meqIlOO grams)
4. Comparison of foreign soil/sediment (if used), including soil/sediment characteristics (noted above)
and microbial activity, to U.S. soils/sediments
5. Description of soil/sediment preparation including sieve size
6. Description of water source and characteristics (including pH, conductivity, hardness, etc) if available
7. Indicate other studies supporting this registration which use this soil/sediment
D Test Methodology
C-305
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Subdivision N
Guideline Ref. No. 162-3
December 24, 1989
1. Level of fortification
2. Description of treatment technique
3. Temperature range during experiment
4. Description of method used to initiate and maintain anaerobic conditions
5. Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
6. Description of volatility trap and analysis procedures (if used)
7. Description of extraction method for parent material and individual degradates in soil/sediment and
water fractions
8. Description of extraction efficiency as percent of applied individually for parent and degradates
9. Description of analytical method including standards used and percent recovery
10. Descnption of storage conditions
ii. Description of storage stability and results
E. Reported Results
1. Description of material balance at sampling intervals as percent of applied
2. Description of half-life estimation for parent
3. Description or graph of formation and decline of parent and degradates
4. Description of anaerobicity measurement (Eh) and results if measured.
F. Discussion of Results
C-306
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Subdivision N
Guideline Ref. No. 162-4
December 24, 1989
162-4 Aerobic Aquatic Metabolism
ACCEPTANCE CRiTERIA
Does your study meet the following a eptan aiteria?
1. — Test substance was the Technical Grade (TGAJ) or a Pure Active Ingredient Radiolabeled
(PAIRA).
2. — Radiopurity and specific activity of Active Ingredient (AI)(if radiolabeled).
3. — The positions of the Al radiolabeling were appropriate (if radiolabeled).
4. — Experiments were conducted with each respectively labelled ring (for compounds containing
more than one ring structure)
5 _ Water was adequately characterized.
6. — Foreign soils/sediment (if any) were adequately compared with domestic (USA)
soils/sediments.
7. — Sediment as representative of the intended use site.
8 — Both aerobic and aquatic conditions were assured and maintained
9 — Application rate was consistent with the sensitivity of the analytical method, so that
degradates could be identified.
10 — Reported expenmental temperature was held constant (± 1°C) berween 18 and 30°C.
ii — Sampling intervals were adequate over the 30 days of the study.
12 — A reasonable attempt was made to identify the parent and all degradates [ which exceeded
10% of application or 0.01 ppm (whichever is smaller)).
13. — A reasonable attempt was made -- perhaps with multiple solvent systems •- to extract
metaboites/degradates.
14. — Half-life of parent was calculated.
15 — Patterns of formation and decline of major degradates were reported.
16. — Material balance was reasonable (>90% - <110%)
Criteria marked with a * are supplemental and may not be required for every study.
C-307
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Subdivision N
Guideline Ref. No. 162-4
December 24, 1989
17 * Detection limits were reported.
18. Appropnate analytical methods were provided.
Criteria marked with a * are supplemental and may not be required for every study.
C-308
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Subdivision N
Guideline Ref. No. 162-4
December 24, 1989
162-4 Aerobic Aquatic Metabolism
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Condusions
B. Test Material
1 The chemical structure: (for radiolabeled substances include “ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3 Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to saiisl this date requirement, that use alternate
radiolabeling and that are included in this summary package.
C Test Media
1 Description of test soil/sediment source (including country, state of origin, etc) for each soil used
2. Description test soil/sediment type (USDA classification)
3. Description of soil/sediment characteristics
a. % sand, silt, clay
b. % organic material
c.pH
d. Cation exchange capacity (meq/100 grams)
4. Comparison of foreign soil/sediment (if used), including soil/sediment characteristics (noted above)
and microbial activity, to U.S. soils/sediments
5. Description of soWsediment preparation including sieve size
6. Descnption of water source and characteristics (including pH, conductivity, hardness, etc) if available
7. Indicate other studies supporting this registration which use this soil/sediment
C-309
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Subdivision N
Guideline Ref. No. 162.4
December 24, 1989
D. Test Methodology
1. Level of fortification
2. Description of treatment technique
3. Temperature range during experiment
4. Description of method used to maintain aerobic conditions
5 Description of soil/sediment and water sampling intervals including time zero concentration:
(indicate hr, day, weeks)
6 Description of volatility traps and analytical procedures (if used)
7. Description of extraction method for parent material and individual degradates in soil/sediment and
water fractions
8. Description of extraction efficiency as percent of applied individually for parent and degradates
9. Description of analytical method including standards used and percent recovery
IC Description of storage conditions
Ii Description of storage stability and results
E. Reported Results
1 Description of material balance at sampling intervals as percent of applied
2. Description of half-life estimation for parent
3. Description or graph of formation and decline of parent and degradates
4. Description of aerobicity measurement (Eh) and results if measured.
F. Discussion of Results
C-310
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Subdivision N
Guideline Ref. No 163-1
December 24, 1989
163-1 Leaching/Adsorption/Desorption
ACCEPTANCE CRITERIA
Does vur study meet the fbllowing a pzan criteria?
1. Testing method was Batch Equilibrium or Soil Column.
2. Test substance was the Technical Grade (TGAI) or a Pure Active Ingredient Radiolabeled
(PAIRA).
3. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeled).
4 — The positions of the A! radiolabeling were appropnate (if radiolabeled).
5. — Experiments were conducted with each respectively labelled ring (for compounds containing
more than one ring structure.
6. — At least 4 soils were used, which had pH’s between 4 and 8.
7 — One soil had a %OM < 1%.
8 — One of the soils was the same type of soil used in the aerobic metabolism study.
9. — Soils were completely charactenzed using the USDA classification system.
10 — Soils were not sieved using a screen smaller than 2mm.
11 _ Soil moisture was maintained at 75% of 113 bar dunng the aging process.
12. — Foreign soils (if any) were adequately compared with domestic (USA) soils.
13. Test substance was added to the soil at the highest recommended label rate for a single
application.
14 * Material balance was reasonable (>90% - <110%).
15 — Mobility of parent and all major degradates identified in the laboratory metabolism studies
was determined during each column leaching study.
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-31 1
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Subdivision N
Guideline Ref. No. 163-1
December 24, 1989
For SOIL TLC... (no longer recommended)
1. — Reference standards for degradates identified in the laboratory metabolism studies were used.
2. — R values were reported for parent and degradates.
For SOIL COLUMN...
1. — Elution volume was equivalent to 20 (508 cm times the cross-sectional area of the column).
2. Column length was adequate.
3. — Distribution (identity and quantity) of major degradates/metabolites within the column
(including eluate) was determined.
For AGED LEACHING...
1. — Test substance was aged for 30 days or one half-life (whichever is shorter).
2. A reasonable attempt made to identify the parent and degradates.
For BATCH EQUILIBRIUM...
1. — Test substance was equilibrated at a minimum of 4 concentrations, with adequate time
allowed for complete equilibration.
2. — 0.OIN Ca solution was used.
3. — Appropnate analytical methods were provided.
4. — Detection limits were reported.
5. — K , K and K were reported.
Criteria marked with a * are supplemental and may not be required for every study.
C-312
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Subdivision N
Guideline Ref. No. 163-1
December 24, 1989
163-1 Leaching/Adsorplion/Desorpijon
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of Condusions
B. Test Material
1. The chemical structure: (for radiolabeled substances include ‘*‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3. Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisf i this data requirement, that use alternate
radiolabeling and that are included in tius summary package.
C. Descriptions of the Required Four Soils
1 Description of test soil source (including country, state of origin, etc) for each soil used
2. Description test soil type (USDA classification)
3. Description of soil characteristics
a. % sand, silt, clay
b. % organic material
c.pH
d. Cation exchange capacity (nieq/100 grams)
4 Comparison of foreign sod (if used), including soil charactenstics (noted above) and microbial
activity, to U.S. soils
5. Description of soil preparation including sieve size and sterilization technique (if used)
6. For aged residues, description of aerobic incubation conditions prior to leaching test
7. Indicate other studies supporting this registration which use this soil
C-313
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Subdivision N
Guideline Ref. No. 163-1
December 24, 1989
D. Test Methodology
1. Batch equilibrium method
a. Levels of fortification
b. Description of equilibrating solution (e.g. (lOIN Ca ion solution)
c. Description of equilibration time and technique
d. Description of cosolvent, if used
e. Description of sampling intervals including time zero concentration: (indicate hr, day)
f Description of analytical method for parent and degradates
2. Soil column
a. Description of column (dia. x length) and soil column preparation
b Description of column equilibration, if any
c. Description of and volume of soil column eluate
d. Description of extraction method for parent material and individual degradates
e. Description of analytical method for parent and degradates
3 Soil Thin Layer Chromatography (TLC)
a. Description of TLC plate preparation
b, Description of test substance application technique
E. Reported Results
1. Batch equilibrium results
a. Description of material balance at sampling intervals as percent of applied
b. Description of K (and K , if available) calculations and results
C-3 14
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Subdivision N
Guideline Ref. No. 163-1
December 24, 1989
2. Soil column
a. Description of material balance as percent of fortification levels
b. Description of distribution curve of parent and degTadates in 6 inch (15 cm) soil column segments
and in the eluate
3. Soil TLC
a. Description of material balance as percent of fortification levels
b Description of soil TLC R( calculations and results according to mobility class of the parent and
individual degradates
C-315
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Subdivision N
Guideline Ref. No. 163-2
December 24, 1989
163-2 Laboratoiy Volatility
ACCEPTANCE CRIThRIA
Does your study meet the following acceptance criteria?
1 _ A greenhouse or field volatility study was provided as a substitute for this data requirement.
2. — A typical End Use Product (EP) was used, or an adequate explanation provided to justify the
alternative chosen.
3. — Radiopurity and specific activity of Active Ingredient (Al) (if radiolabeted).
4. The positions of the Al radiolabeling were appropriate (if radiolabeled).
5. Experiments were conducted with each respectively labelled ring (for compounds containing
more than one nng structure).
6. Soils were completely charactenzed, using the USDA classification system.
7. Foreign soils (if any) were adequately compared with domestic (USA) soils.
8. Test substance was added to the soil at the highest recommended label rate for a singic
application.
9. — Sampling intervals were adequate.
10. — The decline of the AL was clearly established.
11. — Material balance was reasonable (>90% - <110%)
12. Trapping efficiencies were reasonable.
13. — Reported experimental temperature was held constant (±1°C) between 18 and 30°C.
14._ Volatility was reported in the correct units (j g/cm 2 /hr).
15 — The concentration of the Al in air was measured.
16.*_ The concentration of AL in air was reported in the correct units (ng/m 3 or
17._ The vapor pressure was reported in the correct units (Torr, or mm Hg), including the
temperature at which it was determined.
18. — Appropnate analytical methods were provided.
Criteria marked with a * are supplemental and may not be required for every study.
C-316
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Subdivision N
Guideline Ref. No. 163-2
December 24, 1989
19._ Detection limits were reported.
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-317
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Subdivision N
Guideline Ref. No. 163-2
December 24, 1989
163-2 Laboratory Volatility
GUIDANCE FOR SUMMARIZING STUDIES
Items in snmmary should include the items discussed in Chapter 2 of this package and the specific items
listed below.
A. Summary of CondusloiLs
B Test Material
1. The chemical structure: (for radiolabeled substances include ‘‘ to indicate radiolabel position, if
radioactive compound is used)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity,
if radioactive compound is used)
3. Vapor pressure at 25°C and water solubility of the test material (include temperature and pH)
4 Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package
C Soil Description
1. Description of test soil source (including country, state of origin, etc) for each soil used
2 Description test soil type (USDA classification)
3. Description of soil characteristics
a. % sand, silt, clay
b. % organic material
c.pH
d. Cation exchange capacity (meq/100 grams)
4. Comparison of foreign soil (if used), including soil characteristics (noted above) and microbial
activity, to U.S. soils
5. Description of soil preparation including sieve size and sterilization technique (if used)
6. Indicate other studies supporting this registration which use this soil
C-318
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Subdivision N
Guideline Ref. No 163.2
December 24, 1989
D Test Methodology
1. Level of fortification
2 Description of treatment technique and laboratory rest equipment
3. Description of soil moisture content throughout study
4. Temperature range during experiment
5. Description of method used to maintain aerobic soil conditions and collect air samples
6 Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
7. Description of volatility traps and analysis procedures (if used)
8. Description of analytical method including standards used and percent recovery
E. Reported Results
1. Description of calculations for volatility expressed as j g/cm 2 /hour, air concentration expressed as
g/m 3 of mg/rn 3 and vapor pressure expressed as Torr (or other equivalent units)
2 Description of results based on calculations
F Discussion of Results
C-319
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Subdivision N
Guideline Ref. No. 163.3
December 24, 1989
163-3 Field Volatility
ACCEPTANCE CRI’IER1A
Does your study m t the following aa ptance aiteria?
I — The study was conducted domestically (USA).
2. — A typical End Use Product (EP) was used, or an adequate explanation provided to Jusu& the
alternative chosen.
3. Test substance was added to the soil at the highest recommended label rate for a single
application.
4. — The test site (including soil type) was typical of actual use; soil was adequately characterized
using the USDA classification system.
5. The site used for this study was clearly shown to have no previous use history involving this
(or closely related) compound (Al).
6. — Environmental conditions were typical, and adequately described.
7 — Sampling intervals were adequate.
8 — The decline of the Al was clearly established.
9 s Volatility was reported in the correct units ( Lg/cm 2 /hr).
10. — The concentration of the Al in air was measured.
11 _ The concentration of Al in air was reported in the correct units (ng/m 3 or Lgfm 3 ).
12.*_ The vapor pressure was reported in the correct units (Torr, or mm Hg), including the
temperature range over which it was determined.
13 — Appropriate analytical methods were provided.
14.*_ Detection limits were reported.
15. — A storage stability study was conducted using either spiked field or spiked laboratory samples
to determine the stability of samples under typical storage conditions.
Criteria marked with a * are supplemental and may not be required for every study.
C -320
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Subdivision N
Guideline Ref. No. 163.3
December 24, 1989
163-3 Field Volatility
GUIDANCE FOR SUMMARiZING STUDIES
Items in summary should indude the items discussed in C apter 2 of this package and the specific items
listed below.
A. Summary of Conclusions
B. Test Material
1. The chemical structure: (for radiolabeled substances include ‘0’ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3 Volatility and water solubility of the test material (include temperature range and p1-I)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package.
C. Test Site
1. Location
2 Description of test soil type (USDA classification)
3. Descnption of soil charactensties
a. % sand, silt, clay
b. % organic material
c.pH
d. Cation exchange capacity (meq/100 grams)
4. Indicate other studies supporting this registration which use this soil
5. Descnption of environmental conditions during test
D. Test Methodology
1. Application method/rate
2. Descnption of method used to collect air samples
3 Description of sampling intervals including time zero concentration: (indicate hr, day, weeks)
C-321
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Subdivision N
Guideline Ref. o. 163-3
December 24, 1989
4. Description of volatility traps and sampling procedures
5. Description of analytical method including standards used and percent recovery
E. Reported Results
1. Description of calculations for volatility expressed as .LgJcm 2 ,bour, air concentration expressed as
Lg/m 3 of mg/rn 3 and vapor pressure expressed as torr (or other equivalent units)
2. Descnption of results based on calculations
3. Description of climatic conditions during study
F Discussion of Results
C-322
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Subdivision N
Guideline Ref. No 164-1
December 24, 1989
164-1 Terrestrial Field Dissipation
ACCEFFANCE CRITERIA
Does your study meet the following a ptance eriteria?
1 — The study was conducted domestically (USA).
2 — The site used for this study was clearly shown to have no previous use history invclving this
(or closely related) compound (Al) or was clearly shown to contain no background residues
or analytical interferences.
3. — The test site (including soil type) was typical of the proposed use pattern. If intended
application is to turf, vineyard, orchard or other similar use patterns, a bare-ground plot is
also required.
4. — Study was conducted under typical use conditions.
5. — At least 2 sites were used, either in 2 different areas representative of the intended usage, or
in 1-2 similar sites for limited use patterns.
6. — A typical End Use Product (EP) was used, or an adequate explanation provided to justify the
alternative chosen.
7 Test substance was added to the soil at the highest recommended label rate for a single
application the highest recommended rate for each application or multiple application.
8 _ Specifics were provided regarding time of year, meteorological conditions dunng exposure,
etc.
9 — Soil was adequately characterized using the USDA classification system.
10. — Samples were selected randomly.
11. — The soil cores were of sufficient number with minimal compositing for each sampling time to
define the heterogeneity of the soil and pesticide degradation. A minimum of 15 soil cores
with a minimum of three composites is recommended per plot
12. — Sampling was done to a sufficient depth to define leaching (90 cm) at all sampling intervals,
a rationale was given for not doing so.
13. — Monitoring adequately detected all major degradates identified in the aerobic and/or
anaerobic metabolism studies.
Criteria marked with a are supplemental and may not be required for every study.
C.323
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Subdivision N
Guideline Ref. No. 164-I
December 24, 1989
14 — Study was conducted until patterns of decline of parent and patterns of formation/decline of
degradates were established.
15._ The depth to the water table was reported.
16. — Sampling intervals were adequate to define t under field conditions.
l7. — Sampling intervals were adequate to track any vertical movement associated with rainfall.
18. — Appropriate analytical methods were provided.
19 — Half-life of parent and major degradates under field conditions was determined.
20. A storage stability study was conducted using either spiked field or spiked laboratory samples
to determine the stability of samples under typical lab storage conditions.
21. — The test duration following a single event was consistent with the following:
Field and vegetable crop uses: 18 months
Orchard crop and pastureland uses: 12 months
Domestic outdoor, park, ornamental, and turf uses: 4 months
Rights of way, shelter belts and related uses: 2 months
22 — Detection limits were reported
23 A reasonable attempt was made to identify the parent and all degradates [ which exceeded
10% (yield) of the initial concentration of the Al or 0.01 ppm (whichever is smaller)J.
Criteria marked with a * are supplemental and may not be required for every study.
C-324
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Subdivision N
Guideline Ref. No. 164-1
December 24, 1989
164-1 Terrestrial Field Dissipation
GUIDANCE FOR SUMMARIZING STUDIES
Items in summary should include the items discussed in (]iaptcr 2 of this package and the specific items
listed below.
A. Sununaiy of Conclusions
B. Test Material
1. The chemical structure: (for radiolabeled substances include “ to indicate label position)
2. The punty of the test substance (for radiolabeled substances include radiopunty and specific activity)
3. Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package.
C. Description of the required t test sites
1. Identify each test location (including state, county etc) for each test site used
2. Description test soils type (USDA classification)
3 Description of soil characteristics
a. % sand, silt, clay
b. % organic material
c.pH
d. Cation exchange capacity (meq/100 grams)
4. Indicate other studies supporting this registration which use this soil
5. Description of environmental conditions during test
6. Description of pesticide use history of site, if available
7. Topographical description of test sites.
C-325
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Subdivision N
Guideline Ref. No. 164-1
December 24, 1989
D Test Methodology
1. Description of application method/rate and other field test data
2. Description of sampling methodology and sampling intervals including pre-appltcauon, date of
application and immediate post-application (indicate hr, day, weeks)
3 Description of analytical method of soil cores including standards used and percent recovery
4. Description of soil profile and depth to water table, if available
5. Description of sample storage conditions
6. Description of storage stability and results
7. Description of climatic conditions, including amount of rainfall or imgation water, during test
period
E. Reported Results
1. Description of half-life calculation for parent and degradates
2. Description or graph of formation and decline of parent and degradates
3 Description of leaching movement, if observed
F Discussion of Rcsulls
C-326
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Subdivision N
Guideline Ref. No. 164.2
December 24, 1989
164-2 Aquatic Field Dissipation
ACCEPTANCE CRITERIA
Does vur study meet the following a ptanoe criteria?
1. — The study was conducted domestically (USA).
2. — A typical End Use Product (EP) was used, or an adequate explanation provided to justify the
alternative chosen.
3 — At least 2 sites were used, either in 2 different areas representative of the intended usage, or
in 1 to 2 similar sites for limited use patterns.
4. Test substance was added to the soil at the highest recommended label rate for a single
application or the highest recommended rate for multiple applications.
5. — The test site (including soil type) was typical of the proposed use pattern.
6. — The site used for this study was clearly demonstrated to have no previous use history
involving this (or closely related) compound (A!) or was clearly shown to contain no
background residues or analytical interferences.
7. — Study was conducted under typical use conditions.
8 *_ Specifics were provided regarding time of year, meteorological conditions during exposure,
etc.
9. — Soil and sediments (if required) were adequately characterized using the USDA classification
system.
10. — Soil, water and sediments (if required) were analyzed.
11 — Samples were selected randomly.
12. — The soil cores were of sufficient number with minimal compositing for each sampling time to
define the heterogeneity of the soil and pesticide degradation. A minimum of 15 soil cores
with a minimum of three composites is recommended per plot.
13. — Soil sampling was done to a depth of at least 15 cm.
14. — Sediment sampling was done to a depth of at least 5 cm.
15. — Sampling intervals were adequate to define t of parent under field conditions.
Criteria marked with a * are supplemental and may not be required for every study.
C..327
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16. The test duration following a single event was consistent with the following table:
water 1 month
sediments 6 months
9 months
6 months
9 months
(for Aquatic non-crop uses)
(for multiple application)
soil (for Aquatic non-crop uses)
(for multiple applications)
Appropriate analytical methods were provided.
Detection limits were reported.
— A reasonable attempt was made to identify the parent and all degradates [ which exceed 10%
(yield) of the initial concentration of the Al or 0.01 ppm (whichever is smaller)J.
Half-life of parent and major degradates under field conditions was determined.
A storage stability study was conducted using either spiked field or spiked laboratory samples
to determine the stability of samples under typical storage conditions.
Subdivision N
Guideline Ref. No. 164-2
December 24, 1989
17
18. $
19.
20.
21.
Criteria marked with a * are supplemental and may not be required for every study.
C-328
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Subdivision N
Guideline Ref. No. 164.2
December 24, 1989
164-2 Aquatic Field Dissipation
GUIDANCE FOR SUMMARIZING STUDIES
Items ui summary should include the items discussed in ( apter2 of this package and the specific items
listed below.
A. Summary of Conclusions
B. Test Material
1. The chemical structure: (for racliolabeled substances include ‘‘ to indicate label position)
2. The purity of the test substance (for radiolabeled substances include radiopurity and specific activity)
3 Solubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this summary package
C. Description of the Required The Test Sites
1. Identify each test location (including state, county etc) for each test site used
2. Description test soil and sediment type (USDA classification) and water characteristics (pH,
hardness, sediment load, etc)
3. Description of soil and sediment characteristics
a. % sand, silt, clay
b. % organic material
c. pH
d. Cation exchange capacity (meqIIOO grams)
4. Indicate other studies supporting this registration which use this soil and sediment
5. Description of environmental conditions during test
6. Descnption of pesticide use history of site, if available
7 Topographical description of test sites.
C-329
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Subdivision N
Guideline Ref. No. 164-2
December 24, 1989
D. Test Methodolo ,
1. Description of application method/rate and other field test data
2. Description of sampling methodology and sampling intervals including pre-application, date of
application and immediate post-application to maximum test duration (indicate hr, day, weeks)
3. Description of analytical method of soil/sediment and water samples including standards used and
percent recovery
4 Description of sample storage conditions
5. Description of storage stability and results
6. Description of climatic conditions, including amount of rainfall or flood water, during test period
7. Description of flow data in terms of volume or linear flow
E. Reported Results
1. Description of half-life calculation for parent and degradates
2 Description or graph of formation and decline of parent and degradates
F Discussion of Results
C-330
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Subdivision N
Guideline Ref. No. 164-3
December 24, 1989
164-3 Forest Field Dissipation
ACCEF ANCE CRITERIA
Does your study meet the killowing areptanee aiteria?
1. — The study was conducted domestically (USA).
2. — A typical End Use Product (EP) was used, or an adequate explanation provided to justify
the alternative chosen.
3. — Test substance was applied at the highest recommended label rate for a single application or
the highest recommended rate for multiple applications.
4 — Application rate was confirmed by immediate post-application sampling.
5 — The method of application was consistent with label requirements/recommendations.
6 — The test site (including soil type) was typical of the proposed use pattern.
7. The site used for this study was clearly demonstrated to have no previous use history
involving this (or closely related) compound (Al) or was clearly shown to contain no
background residues or analytical interferences.
8 — Soil and sediments were adequately characterized using the USDA classification system
9 — Samples were taken of each of the following components:
- foliage (if foliarly applied)
- leaf litter
- soil under leaf litter
- exposed soil
- standing (pond) water
- moving (stream) water
- sediments (ponds)
- sediments (stream)
10. — The correct sampling intervals were used for...
- foliage’
- leaf litter’
- soil under leaf litter’
- exposed soil’
- standing (pond) wate?’
- moving (stream) waterb
Criteria marked with a * are supplemental and may not be required for every study.
C-331
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Subdivision N
Guideline Ref. No. 164-3
December 24, 1989
- sediments (pond)b
- sediments (stream)b
11. — Samples were taken for an adequate duration for...
- foliage (12 moc)
- leaf litter (12 mcf)
- soil under leaf litter (12 moc)
- exposed soil (12 moc)
- standing (pond) water (1 moc)
- moving (stream) water (imoc)
- sediments (pond and stream) (6 moc)
12. — Monitoring adequately detected the major degradates identified in the aerobic and/or
anaerobic metabolism studies.
13 Half-life of parent and major degradates under field conditions was determined.
14 — Specifics were provided regarding time of year, meteorological conditions during exposure,
etc.
15. Appropriate analytical methods were provided
16.*............. Detection limits were reported.
17 — A storage stability study was conducted using either spiked field or spiked laboratory samples
to determine the stability of samples under typical lab storage conditions.
a pre-application, day 0, and at least 3 samples within the first week for each single or multiple
application.
pre-application, day 0, and at least 1 sample within the first week for each single or multiple
application.
or until establishment of patterns of decline of parent, and formation and decline of degradates.
Criteria marked with a * are supplemental and may not be required for every study.
C-332
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Subdivision N
Guideline Ref. No 164-3
December 24, 1989
164-3 Forest Field Dissipation
GUIDANCE FOR SUMMARIZING SIIJDLES
Items in summary should include the items discussed in Chapter 2 of this package and the specilic items
listed below.
A. Summavy of Conclusions
B. Test Material
1. The chemical structure of the test substance
2. The purity of the test substance and pounds Al per acre applied
3. Solubility of the test matenal in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement and that are included
in this summary package
C. Description of the Test Site
1 Identify test locaticn (including country, state of origin, etc)
2. Description test soil type (USDA classification)
3 Descriptton of soil characteristics for exposed soil areas
a. % sand, silt, clay
b. % organic matenal
c. pH
d. Cation exchange capacity (meq/100 grams)
4 Indicate other forest dissipation studies supporting this registration
5. Description of climatic conditions during test
6. Descnption of pesticide use history of site, if available
7. Topographical description of test site
C-333
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Subdivision N
Guideline Ref. No. 164-3
December 24, 1989
D. Test Methodology
1. Description of application method/rate and other field test data
2. Description of sampling methodology for the vanous environmental components (e.g. foliage, leaf
litter, covered and exposed soil, standing and moving water and sediments)
3. Description of sampling intervals including pre-application, date of application and immediate post-
application for the test duration (indicate hr, day, weeks)
4. Description of methods of analysis of the various environmental components of the study including
standards used and percent recovery
5 Description of sample storage conditions
6. Description of storage stability and results
7. Description of climatic conditions, including amount of rainfall or irrigation water, during test
period
E Reported Results
I Description of half-life calculation for parent and degradates for each substrate analyzed (e g. foliage,
leaf litter, etc)
2. Description or graph of formation and decline of parent and degradates
3. Description of other field test data
F. Discussion of Results
C-334
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Subdivision N
Guideline Ref. No. 164-5
December 24, 1989
164-5 Lrnig-Term Soil Dissipation
ACCEPTANCE CRITERIA
Does )Thir study meet the following aeeeptanee aiteria?
I. — The study was conducted domestically (USA).
2 — The site used for this study was clearly shown to have no previous use history involving this
(or closely related) compound (Al) or was clearly shown to contain no background residues
or analytical interferences.
3 — The test site (including soil type) was typical of the proposed use pattern. If intended
application is to turf’, vineyard, orchard or other similar use patterns, a bare-ground plot is
also required.
4. — Study was conducted under typical use conditions.
5 — At least 2 sites were used, either in 2 different areas representative of the intended usage, or
in 1-2 similar sites for limited use patterns.
6 — A typical End Use Product (EP) was used, or an adequate explanation provided to justify the
alternative chosen.
7. — Test substance was added to the soil at the highest recommended label rate for a single
application or the highest recommended rate for multiple applications.
8 _ Specifics were provided regarding time of year, meteorological conditions during exposure etc.
9. — Soil was adequately characterized using the USDA classification system.
10. Samples were selected randomly.
11. — The soil cores were of sufficient number with minimal compositing for each sampling time to
define the heterogeneity of the soil and pesticide degradation. A minimum of 15 soil cores
with a minimum of three composites is recommended per plot.
12. — Sampling was done to a sufficient depth to define leaching (90 cm) at all sampling intervals,
or a rationale was given for not doing so.
13. — Monitoring adequately detected all major degradates identified in the aerobic and/or
anaerobic metabolism studies.
Criteria marked with a ° are supplemental and may not be required for every study.
C-335
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Subdivision N
Guideline Ref. No. 164-5
December 24, 1989
14. — Study was conducted until patterns of decline of parent and patterns of formation/decline of
degradates were established.
15._ The depth to the water table was reported.
16. — Sampling intervals were adequate to define t under field conditions.
17. — Sampling intervals were adequate to track any vertical movement associated with rainfall.
18. — Appropriate analytical methods were provided.
19. — Half-life of parent and major degradates under field conditions was determined.
20 — A storage stability study was conducted using either spiked field or spiked laboratory samples
to determine the stability of samples under typical storage conditions.
Criteria marked with a * are supplemental and may not be required for every study.
C-336
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Subdivision N
Guideline Ref. No. 164-3
December 24, 1989
164-S Lo g-Tcrm Sod Dhe patâo
GUIDANCE FOR SUMMARIZING STUDIES
e thsm nary s uId I i the itc dir ’ Al iii C iqa 201 thie ps e and the s flc i1en
A. Snmm vy of Coadndo
B. Matertal
I The chemical structure of the test substance
2. The purity of the test substance and pounds Al per acre applied
3. Reference additional studies that are presented to satisfy this data requirement and that are included
in this summary package
C. ipuon of the Test &tes
I. Identify each test location (including state, county, etc.) for each test site used
2. Descr puon test soils type (USDA classification)
3 Description of soil characteristies
a. “ sand, sill, clay
b % organic material
c. pH
d. Cation exchange espaaly (meq/lOO grams)
4. Indicate otheT studies supporting thii registration which use this soil
5 Descript 01 vtroum nial nditions during test
6. Descrlpda ci pid 4e use history of site, if available
7. Topograph l atpdon of test sues.
C.337
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Subdivision
Guideline Ret, No. 164-.
December 24. 1989
D. Test 1kD4 1’
1. DescrIption of application method/rate and other geld test data
2. Description of sampling methodology and sampling intervals rot the test duration application, date
of application and immediate pc$t.applicmtion: (indicate hour, day, week)
3. Description of anal method of soil rm including standards ILsed and percent recover i
4. Descnption of soil pro le and depth to water table, if available
5. Description of sample storage conditions
6. Description of storage stability and results
7. Descnpnon of climatic conditions, including amount of rainfall or irrigation water, dunng test
penod
E. Reported Rails
1. Description of half-life calculation for parent and degradates
2. Description or graph of formation and decline of parent and degradates
3 Description of leaching movement. if observed
F ’ D Ion o( Results
C.338
-------
Subdivision N
Guideline Ref. No. 165-I
December 24. 1989
165-1. Co fln Rotadooal Czu
AcW’TANcE R ERIA
Dt ur study me the ti1Iou1ng Iaa a’ftetla?
1. — Test sub Unce was added to the soil at the highest recommended label rate for a single
appticeuon or the highest recommended rate (or multiple applications.
2. — Test substance was a Pure Active Ingredient Radiotabeled (PAIRA).
3 — Radiopunry and speci& activity of Active Ingredient (Al) (if radlolabeled).
4 — The positions of the Al radiolabeling were appropriate (if radiotabeled).
5 — Experiments were conducted with each respectively labeled ring (for compounds containing
more than one ring structure).
6. — A sandy loam soil was used, or another type was justified.
7 — Soil was adequately characterized using the USDA classification system.
8 — Rainfaairrtgation data were provided (it study was outdoor-small plot).
9 All three uop groupinp were used (small grain, leafy vegetable, root crop)
10 — Representative crops were pLanted at appropriate soil aging intervals teg. 30. 120. 210 and/or
360 days).
I i Analyses were performed on appropriate plant parts. with adequate residue characteriz iion
12. — The extraction vheme was adequately descnbed, including all methodolo .
13.’ MaterIal balan was r onable (>90%. <110%).
14 Soil sMtyumd at the appropriate intervals, especially on lay 0 (pre/posi application, and at
the . of pIaahInç each crop.
15 — A,. ,iti1e ana1yd .1 methods were provided.
16. ’_ De kia limits weze reported.
17 — Extent of uptake of residues under laboratory conditions (including identity and
concentration of parent and/or major degradates) was clearly demonstrated.
Criteria marked with a • are supplemental and may not be required for every study.
C-339
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SubdLv .tjo N
Guideline ReL No. 165.1
December 24, 9B9
165.1 C Ro al Ooçs
GUIDAN POR SUMMARIZING gnJDI
Lte in summazy should h,clude the i 4 c ad ii C apte 2 at thi p ge and the sp & items
listed below.
A. S,nnm ry 01 i ulO
B. Test Mstez *
1. The chemical srrucsure (for radfolabeled substance include “ to indicate label position)
2. The purity of the test substance (for radiolabeled subsianc include radiopurity and specific acuvitv)
3. Solubdiry of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement, that use alternate
radiolabeling and that are included in this SUMM2ry package
C. Test , nMl ,
1 Description of test soil (sandy loam) source or test sire (including country, state of ongin. etc
2. Description of sod charactenscica
a. % sand, silt, clay
b. % organic material
c.pK
d. Cation exchange capa ty (me JlOO grams)
3. Comparison of forelga soil (If ined), indudiag sod cbarwer t1 (noted above) and microbial
activity, to U.S soth
4. Desciipdat 01 s preparation induding sieve size
5. Indicate o supporting this registration which ise this soil
D. Tc M ho e
1. DescriptIon of application method/rate
2. Description of other test parameters (e.g. aging periods pnor to rotational crop planting, identity of
rotational crops, time from application to planting and to haivest of rotational crops, CIC)
C .340
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SUb4LVISIO N
Guideline Ref. No. 165.1
December 24. 1989
3. Description of sampling methodolo and sampling intervals includrng pre .apphcauon. date of
application and immediate post appLicaiion: (indicate hr. day, weeks)
4. Descr puon of analytical methods of soil and rotational crop samples including standards used and
percent recovery
5. Descripuon of sample storage conditions
6. Description of storage stability and results
7 Descnpuon of environmental conditions, including amount of rainfall or imgation water, during test
penod
E. Rcponcd Restha
1. Description of residues including parent and degradates in soil of each sampling interval
2 Description and results of haIf.ilfe calculation for parent and degradates in sod, if available
3. Description and amounts of significant residues in rotadonal crops at each sampling interval
including immature (it consumed by domestic animals or humans) and mature plant stages
F Dtscnsssois of Results
C .341
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Subdw lston
Guideline Ref. No. 165 ..
December 24, 1989
165-2 F1e Ro*atk ssl Crops
A xiANcE c flER1A
Dom oer study meet t Ilt .thg eep*auce attcfla?
I. — The study was anducted domestically (USA).
2. — A typical End Use Product (EP) was used, or an adequate explanation provided to u.sti ’ ihe
alternative chosen.
3. — Test substance was added to the soil at the lughest recommended label race for a single
application (as confirmed by immediate post-application sampling) or the htghesc
recommended race for multiple applications.
4. Test substance was d d to the soil ansistent with label recommendauons/requtremenis.
S — At least 2 sites were used. either in 2 different areas representatwe of the intended usage, or
in 1-2 imtLar sites for Limited use patterns.
6. — The appropriate degradateslmetabolites were adequately detec1ed/quanu ed as indicated itt
thc confined rotauoul aop study.
7 — One of the soils was the same type as in the confuted study.
8. — Soil was adequately characterized using the USDA classification s iem.
The test site (including soil type) was typical of the proposed use pattern.
10. — The site used for thh study was clearly demonstrated to have no previous use history
involving this (or closely related) compound (Al).
11 — Specifica were pTo’ lded reprding time of year, meteorological conditions dunng exposure.
etc.
12. — All three op poupinp were used (small grain, lea vegetable, root crop.)
13. — Pr 1 tI I 5 esops were planted at appropriate soil aging intervals (eg. 30, 120. 210, andior
14. — A.4ps were puLimed on appropriate plant pans, with adequate residue characterization.
15. — The estraction i.rb me was adequately descnbed including all cnethodolo .
Criteria marked with a are supplemental and may not be required for evesy study.
C-342
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Subdmaton t4
Guideline Ref. No. 165-2
December 24, 1989
16 — Soil ana’yzed at the approprtate intervals, especially on day 0 (pre/posI application, and at
the time of planting each crop).
17. — Appropriate ana1yt1 l methods were provided.
18._ Detection limits were reported.
19. — Extent of uptake of residues under field conditions (including identity and concentration of
parent and/or major degradates) was clearly demonstrated.
20. — A storage stability study was conducted using either spiked field or spiked laboratory samples
to determine the stability of samples under typical storage conditions.
Critena marked with a ‘ are suppIe iental and may not be required for every study.
C-343
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Subdivision N
Guideline Ref. No. 165.
December 24. 198
165-2 F R t4r I Oop
GU DANc R R SUMMARIZING STUDIES
Iten In snmI ry skinld Include t 1Ie In pea 2 ol th pm ge and the sp i& ncn
listed below.
A. Sn ” ’y 01
B. Tear Material
1. The chemical structure of the test compound
2. The p rnyof the test substance and pounds Al per acre applied
3. Solubtlfty of the tear material in water (include temperature and pH)
4. Reference additional studies that are presented to saust this data requirement and that are included
ut tins summaty package
C. Desalpdon ol t Reqnlind T Teat Sites
1. Identify each test location ( Including state, county nrc) for each test site used
2. Description of teat soil types (USDA classification)
3. Description of soil characteriatles
a. % sand. silt, clay
b. % organic material
c. pH
d. Cation baar capsclty ( q(1OO grams)
4. Indicate - P I . ypwllag ith registration which ne this soil
5. DescrtpcLE kuitm I nd1ttons during I I
6. Dcscrlpdas pea’ t bletory of site, if available
1. TopographIcal description of teat sites.
D. Teat Mnlbod”W
1. Description of application method/rate
C.344
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Subdivision N
Guideline Ret. No. 165.2.
December 24, 1989
2. DescTipt lon of other test parameters (e.g. aging periods prior to rotational crop planting, identity of
roiauonal crops. time from applkanon to planting and to harvest of rotational crops, etc)
3. Description of sampling methodolo i and sampling intervals including pre -application. date of
application and immediate posi.apphcation: (indicate hr. day, weeks)
4. Description of analytical methods of soil and rotational crop samples including standards used arid
percent recovery
5. Description of sample storage conditions
6 Descnption of storage stability and results
7. Description of climatic conditions, including amount 01 rainfall or irrigation water, during test
period
E Reported Results
1. Descnption of residues including parent and degradates in soil of each sampling interval
2. Description and results of half-life calculation for parent and degradates in soil, if available
3. Description and amounts of siga.tficant residues in rotational crops at each sampling interval
including immature (if consumed by domestic animals or humans) and mature plant stages
F DiSc%L Km Of Resul
C-345
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Subdivision
Guideline Ref. No. 165-
December 24, 1989
16S-3 mn1’dna I D Irrigated Cm
A PTANc IFER1A
Dom ur study - the tkwlng pUn criteria?
1. — The study was conducted domestically (USA).
2. — A typical End Use Product (EP) was used, or an adequate explanation provtded to justify the
alternative chosen.
3. Test substance was added to the soil at the highest recommended label rate for a single
application (as confirmed by immediate posI.application sampling).
4 — Test substance was added to the soil consistent with label recommendauons/requirements.
5. — At least 2 sites were used, either in 2 different areas representative of the intended usage, or
in 1.2 similar sites for limited use patterns.
6. — The site used for this study was clear’y demonstrated to have no previous use history
invoMng this (or closely related) compound (Al) or was clearly shown to contain no
background residues or analytical interferences..
7 — Malyses were performed on appropnace plant parts, with adequate residue characterization
8 — The extraction scheme was adequately described, including all methodolo i.
9 — The appropriate degradates/metabolitea were adequately detectedlquanufied as indicated in
the confined rotational crop study.
10. — Soil was adequately characterized using the USDA classification s tem.
11. — The test site (including sod type) was typical of the proposed use pattern.
12. — Specifies were provided regarding time of year, meieorolo esl conditions during exposure.
etc.
13. — Soil . ..l).tJ at the appropriate intervals, especially on day 0 (pre/posl application, and a
th e p nlth$ each cop .
14. — kr. .iIi . W$Z ID analyzed.
15. — Crope e liTigated *1 the highest rate allowable following good agricultural practices.
16. All three crop grouplisga were used (small grain, leafy vegetable, root crop).
Cnteria marked with a are supplemental and may not be required for every study.
C-346
-------
S bdivi on N
Guidettite Ref. Mo. 165.3
December 24, 1989
17. ApproprIate analytical methods were provtded.
I8._ .. .. Detection limits were reported.
19. — Extent of uptake of residues under field conditions (including identity and concentration of
parent and/or major degradales) was dearly demonstrated.
20. — A storage stability study was conducted using either spiked field or spiked Laboratory samples
to determine the stability of samples under typical storage conditions.
Critena marked with a are supplemental and may not be required for ewry study.
C .347
-------
Subdjvi3t
Guidehne Ret No. 16
December 24. 1989
165-3 tP i in Irrigated Ov
OU1DAN R* SUMMARIZING SIUDIES
Items in summasy s aId indDde the hems d’ I In cbaptex 2 of tb ps ge and the sp flc items
A. SnffiwII of QintiuaMms
B. Tess Matm I
1. The chemical structure of the test substance
2. The purity of the test substance and pounds Al per acre applied
3. Sotubality of the test materIal in water (include temperature and pH)
4. Reference addiuonal studies that are presented to satisfy this date requirement and that are included
Lfl this summaty package
C. DeaaIps e of the Rôi iLfvd T T s Sh
1, Identify each test location (including stare t unty etc) for each test site used
2. Description of test sod types (USDA classification) and water characreristica of the irrigatton waler
(pH, hardness, sediment load, etc)
3 Description of sod characteristica
a. % sand, silt, clay
b. % organic material
C. pH
d. Cation ange capadly (meç(100 grams)
4. Indicate os .vppoting this registration which use this soil
5. DescrIpd d _ .L . .1mftl conditions during rest
6. DescrIption dpu’ ’ - hhwly of she, If available
7. TopographIcal description of test sites.
C . .348
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Subdivision N
Guideline Ret No. 165.3
December 24. 1989
D. Test
I. Descnptio of application method/rate to imgatlon water
2. Description of other test parameters (e.g. identity of !mgaced crops, time from water treatment to
Irrigation and to harvest of i.mgaced crops, flow race and amount of irrigation water applied, eic)
3. Description of sampling methodolo , and sampling interval,s (i.e. hr, day, weeks)
4. Descnption of methods of analysis of soil and imgated crop samples including SLandards used and
percent reavery
5 Description of sample storage conditions
6. Description of storage stability and results
7 Description of climatic conditions, including amount of rainfall or imgazion water, during test
period
E. Reported Results
I Descripno of residues including parent and degradates in soil of each sampling Interval
2. Description and results of halfl.ife calculation for parent and degradates in coil, if available
3 Description and amounts of significant residues in imgated crops at each sampling Intcr al including
immature and mature plant stages
F D ct joG of Results
C-349
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Subdivisiol
Guideline Ret. No. l6
D mber 24. 1989
165-4 Bov wmulatloa In F h
MXEPTAN E I1 1A
Does at — m the ng
— Test substance was the Technical Grade (TGAI) or a Pure Active ingredient Radiolabeled
(PArRA).
2. — Radiopurity and specific activity of Active Ingredient (Al) (if radlolabeled).
3. — The positions of the A! radiolabdling were appropriate (it radiolabeled).
4. — Experiments were conducted with each respectively labelled ring (for compounds containing
more than one ring structure).
5. — The correct fIsh species was used (Bluegill Sunfish or Channel Catfish are preferred).
6 — Study was a flow4hrough type (not static).
7 — The pesticide conccnuation in the water was reasonably constant, and did not exceed 110
the 96 hr LC .
8._ A control group was used.
9 — Fish were exposed br about 28 days. or until an accumulation plateau was reached.
10. — Fish were depurated for about 14 days.
11 — Water was sampled at appropriate intervals.
12. — Fish tissues (edible vs vbcera) were appropriately analyzed at the various stages of
accumulation and deputation.
13. — Extractable residues whkh ezt eded 50 ppb were adequately identified.
14. — The n vt 1me was clearly described, including all methodolO ’.
15. — T t “ as e adequately descnbed.
16. — P .lP* c nUitio’ In H 2 OIflsh were adequately repoitad.
L i. — The le l of deg,ndat In the water was not unreasonably high.
18. — Appropriate analytical methods were provided.
Criteria marked with a • are supplemental and may not be required for every study.
C.350
-------
Subdivtsion N
Guideline Ref. No 165.4
December 24, 1989
19 Detection limits were reported.
20. — Raw data Lfl support of the Final Report are available.
Criteria marked wuh a are suppkmeucai and may not be required for every study.
C-351
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Subdivision
Guideline Ref. M c. 165-4
December 24, 1989
165-4 B 1 ’ mu1athos in Th
GUIDANCI FOR SUMMARiZING STUDIES
Itenis in m iy i 1d tai wIc I Ii dlecuued In ( apt 2 of th ge ned the spei & items
listed bebw.
ft. Snm y of
B. Test Materlel
I. The chemi l structure (for radlolabeled substances include ‘‘ to indicate label position)
2. The purity of the test substance (for radiotabeled substances include radiopurity and specific activity)
3. Solubilily of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement. that may or may not
use alternate radiolabding and that are included iii this summary package
C. Test Mtho *
I Identity and description (weight and length) of fish species
2. Description of study design and pre-aposure equilibration and exposure test conditions
3 Description of water source and characteristit (including pH. conductivity, hardness, dissolved
oxygen. etc.)
4. Description of water and flab sampling intervals including exposure arid deputation periods
5. Description of analytlesi metho for water and fish residues including extraction efficiency and
percent reco ry
a Repoited
I. DescrIpd of In eater and fish tissues (whole body. edible and non-edible tissues) during
the ap pu sad in k tissues during depuraflon period.
2. Deacnpdoa of b4oaJEsauadon esiculations and bioconcenlradon ctors for fish uptake (whole
body, edible sad n.edibIe dsaues) at each sampling period
3. Description o( sample storage conditions prior to analysis
4. DescriptIon of p storage stability study and results
C-352
-------
F. DI3aaaà * o( R ft
C-353
S b41vt toft N
Guid t Re No. 63.4
December 24. 1989
-------
Subdjvisi 0 N
Guideline Ref. No. 165-5
December 24, 1989
165-5 w malet1o II Aquatic No-Thi et Otg i’& .
AQ ’rA icE Il ’ERiA
Does , umi, meet the ilo thg afta ?
I. The study was conducted domesdeally (USA).
2._ A typical End Use Product (EP) was used, or an adequate explanation provided to jusufv the
alternative chosen.
3. Application was appropriate to the intended use pattern.
4. — The site which was used for this study clearly shown to have no previous use hisioiy
involving this (or closely related) compound (Al) or was clearly shown to contain no
background residues or analytical interferences.
5. Fish from bottom, middle and surface feeding patterns were analyzed.
6. Fish were adequately described as to species, size, etc.
7. — Test conditions were adequately described.
8. Water,fish were both analyzed at appropriate Intervals.
9 The extraction scheme was clearty described, including all methodoIo ,.
10. — Sediment was analyzed at appropriate intervals.
11. Sediments were adequately characterized.
12. — The appropriate deçadates were analyzed for.
13. Fists Wanes (edible u. visesra) were appropriately analyzed.
14. Ap,. ilasa analytical methode were provided.
15. Dr — Ihela uare reported.
— PL Il esmes a floes in H h were adequately reported.
17. The test she (induding soil type) was typical of the proposed use pattern.
18. SpecIfies were provided reprdlng time of year, meteorological conditions during exposure,
etc.
Criteria marked with a are supplemental and may not be required for evesy study.
C-354
-------
SUb4MS%Ofl N
Guid ljnt Ref. No. 165.5
December 24, 1989
19. — A s1or ge stabillcy study Was conducted using either spiked field or spiked laboratoty samples
to determine the stabthty of samples under typt t lab storage conditions.
20. — Raw data in support of the Final Report are avadable.
Cntena marked with a are supplemental and may not be required for evely study.
C-355
-------
Subdlv(sion N
Guideline Ref. No. 16,1 -5
December 24, 1989
165-5 D W UI$t* O in MIV$IiC Non-Target Otpn
OUIDANC fOR SU? Q4AR 4O STUDIES
Itcn in m &iy skuid l 1 ucIedc 1k tt lianisv4 in ( apIa 2 of th pscbge aM the spe & 1te
LitM heIo .
A Sn11t111 Ay of C .uod IO
B. Test Materbi
1. The chem i structure
2. The purity of the test substance and pounds Al per acre appited
3. Sotubility of the test material in water (include temperature and pH)
4. Reference additional studies that are presented to satisfy this data requirement and that are included
in this summary package
C. Desalptloa o(1k Reiq.&e T T t Sites
1. Identify the test location (including state, county etc) of each site
2. Descnption test soil and sediment type (USDA classification) and water charaCten5tit (pH.
hardness, sediment load, etc)
3 Descnpuon of sod and sediment characteflstl(s
a. % sand, sill; clay
b. % organic material
c. pH
4. Cation hange capa ty (meq/100 grams)
4. lndimte os vm supporting this registranon which use this soil and sediment
5. Desaip1 n of ,troumsntsl oditions during test
6. Description of pesticide tae hbtory of sites, if available
7. Topographical description of test sites
C.356
-------
Subdw ston N
Guideline Ref. No. 165.5
December 24. 1989
D Test Mc* odoao
1. Description of application methodlrate and other field test data
2. Description of soilisediment, fish and water sampling methodoIo and sampling intervals including
pre-applicatton , date of application and immediate posl.apphcation to ina rnum test duration
(indicate hr. day, weeks)
3. Description of methods of analysis of sotL edixnent, fish (whole body and edible tissue) and water
samples including standards used and percent recovety
4. DescriptIon of sample storage conditions
5. Description of storage stability and results
6. Description of climatic conditions, including amount of rainfall or flood water, during test period
7 Description of flow data in terms of volume or linear flow
E. Reported R vIU
1 Description of residues in water and fish tissues (whole body and edible tissues) during the study
period
2 Description of fish bioconcencranon calculations and bioconcernration factors for fish uptake whole
body and edible) at each sampling period
3 Descnption of sample storage conditions prior to analysis
4 Description of sample storage stability study and results
F Dis of R t
C-357
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C.358
-------
SUBDIVISIoN 0
171-3 Direction For Use . .
17 14(a) Nature Of The Residue -. Plants Study .. . . . . . 363
171.4(b) Nature Of The Residue — Livestock 367
171.4(c) & (d) Residue AnalyticaL Method . .. 369
171.4(e) Storage Stability . . . 372
17 1-4 ( j ) Magnitude of the Residue — Potable Water . . . 374
1714(g) Magnitude at the Residue .- FISh . 376
171-4(h) Magnitude of the Residue .- tmgated Crops 378
17 1-4(i) Magnitude of the Residue — Food Handling . . . 3 )
17 1-4 (j) Meat., Milk, Poultry 1 Egg—Feeding 382
1l1- 4 (j) Meat, Milk, Poultry, Egg—D’ermal Treatment 385
171-4(k) Magnitude of the Residue — Crop Field Trials 387
17 1.4(1) Processed FOOdIFCC4 391
171.5 Reduction of Residues 393
171.6 Proposed Tolerances . . 395
171-7 Reasonable Grounds iii Support of Petition . 397
171-13 Submittal of Analytical Reference Standards . . 399
Relationship between Reregistration Tracking Numbera used in this package
arid Guideline Reference Numbers in the Subdivision 0 Pesticide Assessment Guidelines
Phase 3 Pesticide Assessment
Tracking Guideline Number
Number ___________
Nature of the Residue - Plants 17 (4(a) 171 .4(a)(2)
Nature of the Residue . Livestock 1714(b) 171-4(a)(3)
Residue Analytical Method 171-4(c)&(d) 171.4(b)
Magnitude of the Residue:
Storage Stability 171-4(e) 17 1.4(c)(1)(ii)
Crop Field Triais 171.4(k) 171-4(c)(2)(i-iii)
Processed Food/Feed 17 14( 1) 171-4{c)(2)(iv)
MeatJMi lk/POutt ry/EW 171.4 (j) 171-4(c)(3)
Potable Water 1714(t) 17 1-4(C)(4)
Fish 171 -4(g) 171-4(c)(4)
Imgifed Crops 1714(h) 17 1 -4(c)(4)
Food 1 d1hig 1714(i) 17 1-4(c)(5)
C-359
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Subdivision t
Guideline Ref. No. 111-3
December 24, 1989
171-3 Dti o For Use
AQ rrANC CRrrERIA
The following critena are provtded as guidance to registrants concerning the directions for use that
should appear on product labels for food use chemicals. No summary for this guideline is required.
Each registrant must have provided product Labels to EP4¼ submission of these labels will satisfy the
requirement to summarize 171-3.
es ur study me the folksedng ta a1tcr ?
1. — Each formulation to be used identified with the concentration of active ingredient indicated
(% by weight for solid formulations. pounds active ingredient per gallon for liquids)
2. — All crops which are to be treated with each formulation clearly identified
3. — Tolerance or e mpuon from tolerance proposed or established for each crop on label
4. — Impractical or unrealistic use restrictions excluded (eg . restricting feed use of corn forage not
practical due to high economic value and common practice of feeding to livestock)
5. — Names and quantities of stickers. spreaders. or other adjuvants to be added to spray solution
6. — Field and orchard crop directions
— Application rate in quantity of formulation and pounds active tngredienc per acre
(For row or band treatments indicate tithe rate refers to the area treated or to the
entire field)
— Spray volumes to be used per acre
— Maximum number applications per year or growing season
— Minimum Interval between suceessive applications
—. Minimum interval between last application and harvest (preharvest interval = PHI)
— For orchard crops additional information for full coverage spra (quantity of
fOrmulatiOn end pounds active ingredient per 100 gallons spray) and concentrated
sprays (amount active per acre should be related to tree size)
7 — Aerial. ultra iow lume ( 1JLV) and mist spray directions include spray concentration.
amount of acth’s IngredMnt per acre, and spray volume per acre
8. Postharvest tualpdon directions
— Dosage LapteNed in weight fumigant per volume of storage space or headapace or
we*t fn.. 4p * per unit weIght of commodity treated
Tempeesture premure and duration of exposure specified
C v uy aed airtighteess of containers deactibed
a1iou pcoosdurm and time of aeration specified
— Minimum Interval between suoc ”ivc applications
Criteria marked with a a are supplemental and may not be required for every study.
C-360
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Subdivision 0
Guideline Ref. No 7t .3
December 24, 1989
9. — Anlvul treatment directions
— Concentration of pesii ide in treatment solution
— Quantity of spray, pour-on solution, dust. etc. to be applied per animal
— Amount of time animal to be held in dip tank
— Frequency of treatment
— Mazimuni number of treatments
— PresLaughter interval (interval between last treatment and slaughter) specilied
(interval longer than 3 days usually not considered practical)
to. — Aquatic use directions
— Dosage expressed in quantity of formuLation and pounds active ingredient per surface
acre or parts per million pesticide in the ‘water (in latter case the amount product
per surface area should be related to average pond depth)
— Detailed description provided for specia li2ed equipment
— Minimum distance specified from treated area to potable water or trngatiorl intake
pipe
— If oxygen depletion problems. proportion of pond to be treated and required tnter al
between treatments
— Maximum number of applications per year
— Minimum interval between applications
11 — Food handling establishment use directions
— Type of establishment that may be treated
Dilution instructions and spray concentration
— T pe of application equipment and mode of application (space spray. threct d prav
to crevices, spot treatment. etc.)
— Dosage limitations including cubic and square foot limitations
— Frequency of treatment
— Time of treatment (eg., after-hours in restaurants)
— Instruct ions concerning removal and covering of food. dishes and utensils
— Cleanup proi dures before food processing. preparation or serving resumes
12. — Agricultural premises imiments
— Description of areas to be treated (eg.. fend lot., milking room, animal barn)
— Dosage spc ed (pounds per unit volume treated for fogging applications:
concentration of scuvc ingredient in spray solutions; weight active ingredient per unit
area of feed lot)
— Frequency of treatment
Dtreedo to whether animals should be removed during treatment
Criteria marked with a • are supplemental and may not be required for evety study.
C.361
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S abdMSlO k 0
Owdeliac ReL No. 171
December 24, 1981>
171-3 D(r o Fat U sc
GU1DAN 1 R SUW4ARIZING STUDIES
fteu in —“ ‘y s uId E a4C * tt d sc r4 In ( sptet 2 Lh c ge a ihe sp fr ten
L ted be1o .
No summarf for thAs guideline required.
C.362
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Subdjvts 1o 0
Guide j Ref. No. 17 l-4(a)
December 24, 1989
171-4( 1) Nature Ot The R id c - P n Study
AQ P’rAJ4c -flER1A
ü ur study meet the k11o thg a ptaa afta ?
I, Pesticide radlolabeled in non-labile portion of molecule (Influm label strongly discouraged)
2. — Specz& activity su cienI to permit detccuon of low residue levels (0.01-0.1 ppm range)
3. Radiocherni uy pure grade of active Ingredient
4. Pesticide applied to plant in manner simulating expected use
5. Total radioactivity measured in plant parts used (or human food or animal feed
6. Extractability of residue into solvents determined
7 — Most of radioactivity extracted or exhaustive attempts (acid, base, enayme) made to do so
8. — Major components or portion of the terminal residue identified (preferably by at least two
techniques.c.g., TLC, HPLC, MS)
Cntena marked with a are supplemental and may not be required (or evezy study.
C363
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SubdIvision 0
Guideline Ref. No. 71-4(a
December 24. 98
17 1 -4 (a) Nature Of The P n pi St
OUTDM4C FOR SUMMAR 4O ST1JDI
e in summary should leclude the l1 lecuued In C apeer 2 of Ib ps gc and the sp & items
1. Chemical structure and name of test material with site of radlolabel Indicated.
2. Specific activity of test material and detection limit achieved for treated commodities.
3. Radiochemical purity of test materiaL
4. Description of formuLation and application of test material and comparison of application (rate.
timing in terms of growth stage of crop to expected use of pesticide.
5.7. DescriptIon of pro ures used to quantitate and extract radioactivity. Summarize results in a
that includes for each crop part that has been analyzed the interval in days between the last
application and collection of the sample, the total ppm radioactivity (usually in terms of ppm
equivalents of parent pesticide), the % of the activity extracted into solvent(s), and. if acid, base or
enzymatic hydrolyses were conducted, the % of additional total activity released by these procedures.
& Description of prtxedures used to identify radlolabeled residues. Summarize results in a t ihat
includes for each crop part that has been analyzed the interval in days between the last apph .ation
and collection of the sample, the total ppm radioactivity, and the % of total radioactive residue
(TRR) associated with each identifiable residue. Although the various idenitfied residues may have
been found in several different extracts or fractions of a crop part, the % given in the table should
represent the % of the ] ftft In the entire crop part. information on individual extracts or
chromatographic fractious should be limited to the body of the report
If any information is available on the number and magnitude of components within the untder ttfied
fractions, this shoi 1d be inctuded In the summaty. For mample, if an unidentified residue (32% of
TRR) is known to aus*s of as least 5 discrete components, none of which is >10% of the TRR.
indicate this somehow under thh portion of the summery. Other usefuL information on the
unidentified residue that should be presented in the summary, if known, include the proportions that
are extractable (organic $o*vc5 , waler), the proportions releasable by acid/base or enzymatic
hydrolysea, the pardXatag behavior (organic solvents versus waler).
9. ldendfL l she to4mII petition number, if the study has been used to support a tolerance.
F mpl
1. Chemical structure and name of test material with site of radlolabet indicated.
.methyl .3 -bromoenhIne aho known as ‘HERBICIDE
The 14-C radlolabel was uniformly distributed through the phenyt ring.
C.364
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Subdivision 0
Guideline Ref. No. 171-4(a)
December 24. 1989
2. Specl c activity of test material and detection limit achieved for treated commodities.
Specific activity 19.1 ntCvmg. The detection limits for treated soybean crop parts were 001-
0,03 ppm.
3. Radiochemical purity of test material.
The radiochenucal purity of 14 C.HERBICIDE was 98.5%.
4 Description of formulation and application of tesi material and comparison of application (rate.
timing in terms of growth stage of crop) to expected use of pesticide.
Radiolabeled HERBICIDE was formulated as an emulsifiable concentrate by addition of the
emulsifier and organic solvent used in the commercial product. The formulation was diluted
with water and applied to the soybean foliage at the fourth-trifoliate stage at a rate equivalent
to 025 pounds active ingredient per acre, twice the rate permitted on HERBICIDE labels
Soybeans would typically be ut third to fourth-crifoUae stages when HERBICIDE is apptied
5.7 Description of procedures used to quanutate and extract radioactivity. Total radioactwtues of all
plant parts were determined by combustion of aliquots to 4 C0 . Samples were washed with
hexanc followed by blending and extraction with 20% methanol in acetone. In the case of dry
crop parts. refluxing with iN H was used to release additional residues. The total ppm found
in plant parts and the distribution of the residues in the various solvents are summarized below.
Sample summary table - Total ppm in plant parts and the disinbution of residues in solvents
14-C
% into
% into
%
into
% noi
Crop van
hexane
acetone IN
1-ICI
released
Green plant
2
27.5
75
20
•-
5
Dry leaves
75
13.8
15
44
28
13
D i v sa&s
75
19
10
50
23
17
Pods
75
0.7
3
35
40
22
PH1 prebarvest
interval
in days; 14-C in ppm HERBICIDE equivalents
Exu of the tnd ctMty in soybean seeds was not attempted due to the very low level of
res (
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Subdlv$.sio
GuideLine Ref. No. 171
De zmber 24. 1 . ..,
radJo t resldt e. 4 -bydroxy’ refers to N-methyl-3-bromo-4-b$roxyaniline and desmethyr
refers to 3-bromoanlline.
Sample sumntarv table- Total radioactive residu for vanous crop parts
Cron Dart
Total 14-C
%
of TRR
Parent
4-bydroxy
Desmethvl
IJnideMd
Green plant
27 5 ppm
72%
12%
9%
7%
Dry Leaves
13.8 ppm
24%
39%
11%
26%
Dry staib
2.9 ppm
18%
42%
8%
32%
Pods
0.7 ppm
15%
27%
15%
43%
In all esses about half the unidentified residue was activity that could not be extracted by
solvents or HCL The remainder consisted of very polar materials based on their TLC behavior.
C.366
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Subdivision 0
Guideline Ref. No. 171.4(b)
December 24, 1989
171 .4 (b) Nature O( The Residue — U rock
AQ PTANc CRITERIA
Does vur study meet the üo.tng a a aiterla?
1 . — Peaucide radiolabeted in non-labile portion of molecule (trnium label strongly discouraged)
2. Specific activity sufficient to permit detection of low residue levels (0.01.0.1 ppm range)
3. — Radiochentically pure grade of active ingredient used for d ing
.1 — Pesticide administered orally to animals for at Least three consecutive days (or applied
externally for at least 3 consecutive da using a method of application specified in label
directions)
5. — Mimab not preconditioned by dosing with unlabeled material
6. Animals sacrificed within 24 houri of llnal dose or application
7 — Total radioactivity measured in edible tissues (muscle, fat, liver, ruminant kidney) and milk
or eggs
8. — Extractability of residue into solvents determined
9 — Most of radioactivity extracted or exhaustive attempts (acid, base, enzyme made to do so
L0._ Major ci imponen or portion of the terminal residue identified (preferably by at least two
techniques -e.g , ThC HPLC , MS) in edible tissues and milk or eggs
Cntena marked with a • are supplemental and may not be required for every study.
C.367
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Subdiviston 0
Guideline Ref. 14o. 171-4(b)
December 24, 1989
171-4(b) Nature Of The ResAdue —
OUIDAN FOR SU}.QdARJZNO STUD(ES
lte in snu1 ry sboeId ladale ibe ken 4 cireed In apser 2011kM ps*age and ibe sp fr item
Listed beki .
1. Chemical structure and name of iest material with site of radlolabel indicated.
2. Specific activuy of test material and detection Limit achieved for treated commodities.
3. Radiochenucal punly of test materiaL
4. Description of amma.l diet and dosing procedure including ppm pesticide admtnislered/ingested on
total diet basis (thy weight) (or descnpflon of application procedure if dermal treatment involved).
5. Description of animal diet to dosing (if different than 4) and assurance that pesticide not included
in feed.
6. Method of sacrifice and time in bouts from final dose of radiolabeled inatenaL
7.9 Brief description of procedures used to quanhftate and extract radioactivity. Summarize re uIt in a
t j that includes for each edible tissue and miLk (or eg ) the total ppm radioactivity (usually in
terms of ppm equivalents of parent pesticide), the % of the acuvny extracted into so vent(s). arid. if
acid, base or enz 1natic ttydrol ses were conducted, the % of additional total activity released by these
procedures.
[ 0 Description of pro urea used to Identify radiolabeled residues. Summarize results in a ittat
includes for each edible thiuc and milk (or egp) that has been analyzed the total ppm radioacu ity
and the % of total radioactive residue (TRR) associated with each identifiable residue. (Additional
guidance on tables is provided in the related document OUIDANCE FOR SUMMARIZING
‘NATURE OF THE RESIDUE -PLANTS’ STUDY’.I
Li. Ldeniiflcation of the tolesance petition number, if the study has been used to support a tolerance.
C-368
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Subdivision 0
Ouideline Ref. No. 17 1-4(c)&(d)
December Z4, 1989
17 1 -4(c) & (d) R ldae AM)yd i Meshed
AO YFAN 4 fl ’ERL&
Does ur study 1 the Mbu4ng aiteria?
1. — List of equipment, reagents and standards provided along with U.S. sources/suppliers of same
2. Instrumentation and operating conditions described
3. — Detailed description of each step in procedure to enable use by competent analyst unfamiliar
with method
4 — Discrete response for analyte
5 Control values reasonably low compared to tolerance
6. — Adequate recoveries (generally 70%) obtained for !ortiflestions at the tolerance level
7 — Recoveries don’t vary significantly from sample to sample
3 — Evidence that aged residues extracted by procedure (may reference work in metabolism studies)
9 — Data showing that method releases and recovers bound residues (if latter of toxicological
concern)
to — Requirements for regulatory methods (as opposed to those used only for collecting residue data)
— Does not require untreated commodity as blank
— Does not require internal or procedural standard to correct for recovenes (Aiidiuon of
internal standard in final step just prior to injecuon is aoceptable for calibration of
retention times. However, use of internal standard throughout entire procedure to
correct for re nes is not acceptable unless data are available on numerous samp es of
each mautt to show analyle and internal standard behave identically in each step)
Does not require exotic equipment or reagents
— Reasonably rapid in execution
Speci c to measure residue in presence of other reasonably expected pesticides
S alve in relation to tolerance
Confirmatoiy method available
— Method not dalmed to be Confidential Business Information
— Does not me harardous reagents Ousuflcauon needed for use of benzene as solvent or
dla meshnne as inethylating agent)
Criteria marked with a ‘are supplemental and may not be required for cvei study.
C .369
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Subdivision
Guideline Ref. No. 17 1-4(c)d
December 24, b
171-4(c) * (d) Residue AesIytó.l Method
OU1DANC FOR SU?,4MAR1Zn- o ST JDlPS
Items In summazy *bould t ude the Items 4 ed In C apter2 of Ih and the specific items
listed below.
1-3. Bnef description of key steps and instrumentation used in analyticel method.
4. Copies of representative chromatograms, spectra, etc. for reference standard, untreated samples, and
treated or fortified samples illustrating response for analyze.
5-7 Summary containing the control values and recoveries for tested commodities. (See Table [ (or
sample summary table for analyticel methods). It is preferable for the recovery column to list the
individual values and the average recovery for each commodity. However, if a large number of samples
have been e tmined. it would be satisfactory to list the range of recoveries along with the number of
samples, the average recovery and the standard deviation. If more than one analyte was examined.
additional columns may be added to the table or a separate table generated for each analyte.
8. Discussionlsummazy of data showing aged residues extracted by method.
9 Discussion/summary of data showing that bound or conjugated residues of to cologica1 concern are
measured by method. If specific conjugates were added to samples and recoveries measured, the result!
could be included in the summ iy table for items 5.7.
10. For reaulpiprv methods:
- Statement that method does not require (I) untreated commodity as blank. (2) internal standards to
correct for “ covenes, and (3) tic equipment or reagents.
Time (both work hours and total time) required to analyze a sample or set of samples.
• Brief discussion of specificity of method Including results of interference studies.
Detection limit of method, typlesi control values, and comparison of these to tolerances.
- Brief desalpdon of con&ma&oiy method.
- Method i d to be Confidential Business Information.
• State’ method dom not use hazardous reagents or justiflestion for their use.
11. Identiflesdon of the tokna petition number, if the study has been used to support a tolerance.
C. .370
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Subdivtsion 0
Guideline Ref. No. 171-4(c)&d)
December 24. 1989
Table 1. Sample iummuy table o analydcal method
PPM PPM
SamDle Controls EQ % Recovene5 (Avera e
Soybean forage <0.02-0.04 0.05 72,78,83,95 (82)
(green) 0.10 77, 80, 91, 94 (86)
0.50 88, 96 (92)
Soybean nes <0.02-0.05 0.20 68.74,80,84 (77)
(dry) 0.50 11, 81, 87, 90 (82)
Soybeans <0.02 0.02 65, 72. 75, 81 (73)
0.05 75, 78, 91, 97 (85)
0.10 85,94 (90)
Peanuts <0.01.0.02 0.02 70, 75, 80, 82 (77)
0.05 76, 83, 81, 10! (87)
0.20 83, 93, 95 (90)
Peanut hulls <0.02 005 74, 83, 95 (84)
Peanut meal <0.02 005 71. 87 (79)
Peanut oil <0.02 005 81, 97 (92)
C .37l
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Subdiv lzto
Guideline Ret. No. 171-
December 24. 1
171-4(e) StorMe S bUiIy
A TANCE (RrF 1A
Does your study m t the üo frig esa alserla?
L — Sample preparation and fonifi tion described (or in ses where samples with weathered
residues are used, history of p and pesticide treatment provtded)
2. — Storage conditions specified (temperature, containen. form of r.a.c., Ughting, etc.)
3. — Dates Of fortiflatlon (or harvest in e of eld treated samples), pLacement into storage.
sampling, and residue analysis provided
4 — All components of total to dc residue foru( ed into commodity (or present from Eeld treatment
with pesticide) and measured by validated analytical method(s) (description of latter provided or
referenced)
Criteria marked wIth a • are supplemental and may not be required for every study.
C-372
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Subdivision 0
Guideline Ref. No. 171 .4(c)
December 24, 1989
171 -4 (e) Stora SIab lity
GU1DAN( R)R SUMMARIZING STUDIES
Itezzis in summary abould indnde the i1 4W ed In ( aptu 2 at th p ge and the spu flc ite
Listed below.
1. Description of sample treatment prior to pLacement into storage. For field treated crops. description of
mode, rate, and liming at pesticide application. For fortified samples. description of fortification
procedure including form of sample (e.g., whole r.LC., macernie, estract) spiked.
2. Description of form of sample and container placed into storage. Specification of storage temperature
and lighting conditions.
3-4. Brief description of analytical method used to measure residues Including vaudattonlrecovery data.
Summarize in a the residue levels found as a function of time for each commodity, analyte and
fortification level combination. (See Table U for an e ainp1e of such a uble).
Table U. Residue levels
Anatyre PPM PPM after months storage
Conimodirv spiked spiked _Q_ ..J_.. J_.. . ..j .. 12
Corn grain Parent 1.00 0.97 0.86 0.89 0.91 082
Metabl 0.50 etc.
• Meub2 0.50 etc.
Potatoes Parent 0.50 etc.
Metabl 0.20 etc.
• Metab2 0.20 etc.
5. ldennfieaXs of the tolcran petition number, tf the study has been used to support a tolerance
C-373
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SubdivtstOfl 0
Guideline Ref. No. l fl.
December 24. 1
171 .4 ( t) Magnitade of ibm Residne — Potable Water
AC 1’TAN ( rt’ 1A
Does ur stady — lbs ki tng afterle?
I .. — Bodies of water treated a rd1ng to Label directions leading to M TImum residues:
— M if mum applieauon rate and number of appliestions
— Minimum retreatment intervaLs
2. — Data reflect modes of appll tion on label: e.g.. aerial. specialized equipment. treatment of pond
margins only.
3. —— Representative formulations ei nined
4. — Water sampled in center of treated area and at vanoia points to show effect of distance on
residue level
5. — Water sampled on day of treatment and lacer periods to show effect of tune on residue level
6. — Sufficient number of triaLs nducted to determine an appropriate tolerance or ma mum residue
level (factors to txinslder include ‘geographic represenlation’, seasonal and yearly variations. sizes
arid types of bodies of water treated, the toncity of the pesticide)
1. — Total toxic residue measured in water by validated analytleal method (description provided or
referenced)
8. — Description of sample treatment. llection, handling and storage provided (covering periods
from treauncuc to sampling and om sampling thzougb residue analysts)
9 — Dates of treatment, sample llectlon, enuy into storage, and residue analysis
10. — Storage stability data available reflecting storage of water samples (may be referenced to
separate study)
Criteria marked with a • are supplemental and may isot be requited for evezy study.
C . .374
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Subdivision 0
Guideline Ret. No. i7t-4(t)
December 24. t989
171-4(f) Mapit e at the R idne - Po4ab4e Waia
OU1DANC FOR SUMMARIZING SI1JDIES
Items in snm ry s uid indnde the ite dhcuued In C apter 2 at th e and the sp 6 i items
Listed below.
1. Comparison of rate and number of applications and retreatment intervals to those permitted on
product Labels.
2. Comparison of application modes in study to those permitted on product Labels.
3. Description of formulations tested and maximum residues found for each one.
4-5 Description of water sampling procedures with regard to distance from treated area, depth of samples.
and time penods between application and sampling.
6. Discu.ssion of how mals provide adequate estimate of the maximum residue level expected in water
from actual use of the pesticide.
7 Brief description of analytical method used to measure residues including validation/recovery data.
Summarize results of each trial in a table showing the effect of distance and time on residues
S Storage conditions for water samples between sample collection and residue analysis. tSample
tre2tment, collection and handling covered by items 1-5.1
9 Duration ot storage of water samples between collection and residue analysis
10. Summary of data showing stabtlity of residues under storage condinoos and duration similar to those
described in items 8 and 9 (may be referenced to separate study).
It Identification of the tolerance petition number, if the study has been used to support a tolerance.
C-375
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Subdivisiori 0
Oindehne Ref. No. 111-’
December 24,
171.4(g) Mapitede of t P M - F h
Ac zrTAN4 cRrFERIA
Does vur study - L *i kvthg ufter ?
1. — Fish maintained in water containing maximum pesticide residues permitted by product labels
2. — Fish sampled on day of treatment and later periods to show effect of lime on residue level arid
the time required for r dues zo plateau
3. — Residue data obtained for bottom feeders (e g.. catfish) and predators (e.g., bass) in the case of
fish and for moliuses (e.g. clams, oysters) and crustaceans (e.g. shrimp. crabs) in the case of
shellfish
4. — Total toxic residue measured in fish by validated analyiLcal method (description provided or
referen )
5 — Analyses reflect fish comniodny definizion.s in the Pesticzde Analytical Manual. Volume I
6. — If pesticide used in estuanne areas, data provided for whole fish protein concentrate and
smoked. canned or other processed fish products
7. — Description of pesticidal treatment of water and collection, handling and storage of fish samples
provided
8 — Dates of treatment, sample collection, entry Into storage. and residue analysis
9 — Storage stability data available reflecting storage of fish samples between sample collection (i.e.
sacrifice) and residue anal is (may be referenced to separate study)
Criteria marked with a are supplemental and may not be required for eveiy study.
C . .376
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Subdivtsion 0
Guideline Ref. No. 1714(g)
December 24, 1989
t71.4(g) Map.inide o( the R dae — Fbh
GUIDANCE FOR SUP O 4APJZThIO STUDIES
Items in snmin ry heuJd indu the 1tC 4W -ed in ( apt 2 of th p * and the spei fic Ace
listed beIo .
1. Conccrnzauon of pesticide in water and comparison to ma mum level pernuned by product labels.
2. Time periods fish maintained in treated water prior to sampling (or residue analysis.
3 Types of fish and/or shellfish analyzed for residues.
4 Bnef description of analytical method used to measure residues including validationlrecover data
Summarize in a table the total toxic residue found for each type of fish or sheiltl.sh (including control
or untreated samples) as a function of time kept in treated water.
5. Portion of fishlshdflflsh analyzed for residues.
6 Summary of data showing behavior of pesticide residues upon processing of fish.
7 Descrtplion of pesucidal treatment of waler, procedure for collection of fish samples. and storage
conditions of samples.
8 Duration of storage of fish samples between collection and residue analysts.
9 Summary of data showing stability of residues under storage conditions and duration similar to those in
items 7 & 8 (may be referenced to separate study).
10 Identification of the tolerance petition number, if the study has been used to support a tolerance
C.377
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Subdivision
G udeIine Ref. No, l7t--
December 24
171.4(b) Magnitude ot the R - krtpted Crops
AC TAN( ( fl IA
Does your study meet t üou1ng a p aoe aiterti?
1. — Crops irrigated with water antaining marln%um pesticide residues permitted by product labels
2. — Represenutive crops from each crop grouping in 40 CFR 1&).34(f) e mined for residues
resulting from treated irrigation water
3. — Crops sampled ftr residues within one day of irrigation
4. — At least t field trials nducted for each representative crop
5. — Total to c residue measured in crops by validated analysi I method (description provided or
referen )
6. — Descnplion of pesticidal treatment of imption water, irrigation technique, and collecuon.
handling and storage of crop samples provided
7 — Dates of peslicidal treatment of water. irrigation of crops, harvest of crops, entry of crop
samples into storage, and residue analysis
8 — Storage stability data awsilable reflecting storage of crop samples (may be referenced to separate
study)
Cruena marked with a ‘ are supplemental and may not be required for every study.
C-378
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Subdivision 0
Guideline R ef. No 1714(h)
December 24, 19S9
171-4(b) Mag,ün e at the RmIdac - litipted Czo
GUIDANC R)R SUMMARJZU 4G SIIJDIES
hems in snmrn ry ShoUld include the Item dIscussed In ( apier 2 at thIs e and the sp i5c items
Listed below.
1. Concentration of pesticide in water at time of irrigation and companson to macmum level permitted
by product labels.
2. List of crops examined for residues resulting from treated irrigation water.
3 Time periods between irrigation and sampling of crops for residue analysis.
4 Number of trials conducted for each imgated crop
S Brief description of analytical method used to measure residues including validaiionlrecovery data.
Summarize in a table the total to c residue found for each irrigated crop (including control or
untreated samples) harvested within 1-2 days after irrigation. Residues ii samples collecied after longer
irrigation to harvest intervals, if available, should be included iii the table with clear indication of the
observed intervals. -
6 Description of pesiicidal treatment of water, irrigation techniques. and storage conditions of ;amples.
7 Time intervals between pesticidal treatment of waler and imga tion and between har esr of crops and
residue anal%sIS.
8 Summary of data showing stability of residues in crops under storage conditions and duration similar to
those descubed in items 6 and 7 (may be referenced to separate study).
9 Identification of the tolerance petition number, if the siudy has been used to support a tolerance.
C.379
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SubdivisIon 0
Guideline Ret No. lit
Deecmber 24, 1
17 1 -4(I) M*pftn e ol t Mn . — Food I1 fl4 g
A riAN fl LA
Does your study meet the *Iks.*ng i tp4a aiia ?
1. — Pesticide applied in manner leading to highest residues in rood that could result from actual use
(factors to consider include spray concentrauorn type of spray (space. spot.. crack and crevicel;
interval between sprays if multiple appli nons permitted; food covered or uncovered)
2. — Foods representative of those to be present in treated facilities analyzed for pesticide residues
(e.g.. in sc of cafeteria include meat, baked goods. oily food [ butter. cheescj, fresh fruits and
vegetables)
3 — Foods sampled for residue analysis as soon as permuted after application and at various later
tunes
4. — Pesuctde residue measured in representative foods by validated analytical method (description
provided or referenced)
5. — Description of sample treatment, collection, handling, and storage conditions/duration provided
6 — Storage stability data available reflecting storage of food samples from collection until residue
analv is (may be retercnced to separate study)
Criteria marked wfth a ‘ are supplemental and may not be required for evety study.
C .3& l
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Subdivision 0
Guideline Ref. No. 111.4(i)
December 24. 1989
171 -4(1) MapAt e of the Resâdae - Food Handling
GUIDANCE FOR SUMMARIZING STUDIES
in snmrn iy sboeld Indode the tte d c ed in api 2 of th p ge and the spo c items
1&sied bekw.
1. Descnpuon of establishment or room in which study conducted. Descnption of mode and rate of
application and handling of food items (covered ss. unooveTed wrapping rnaienaLs; temperature).
Comparison of pesticide application and food handling to that specified on product labels.
2. List of foods placed in treated area and analyzed for pesticide residues.
3. Time periods elapsed between pesticide application and sampling of foods for residue analysis.
4 Brief descnpuon of analytical method used to measure residues including validationlrecovery data
Residues observed in foods (covered and uncovered listed separately if both analyzed) should be
summarized in a table showing the effect of time (from application to coLlection of samples) on
residues.
5. Storage conditions and duration for food samples between sample collection and residue anaftsis
(Sample treatment. collection and handling covered by items I & 3.)
6 Summary of data showing stability of residues under storage conditions similar to chose descril ed tn
item 5 (may be referenced to separate study).
7 Identification of the tolerance petition number, it the study has been used to support a tolerance
C-38 1
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Subdtv iaior
Guideline Ref. Mo. ill
December 24.
17 1-4W Meet, Milk, Po luy, E —F ’Ing
AC PTANC QII 1A
Does vur study me t küiu4ng pes a1ta i?
1. — Compound(s) fed corresponds to residues expected on feed items
2. — Several dosages mi! cd with one appro maung intake expected from treated feed items, one
or more represenung emggerated intake, and one controL group
3 Multiple animals Included in each dosage group (preferably at least 3 for cattle and 10 for
poultry)
4 Duration of dosing adequate to ensure residues plateau in milk or eggs (preferably at least 4
weeks)
5. — Animals sacrificed within 24 hours of final dose
6. — Total to c residue measured in edible tissues (muscle, fat, liver, cattle kidney) and milk or eggs
of both control and treated animals using validated analytical method (description of latter gwen
or referenced)
7 — Analytical method sufficiently sensitive (0.01-0.05 ppm)
8. — Descnptlon of handling and dosing of animals, feed consumption, and sample collection.
handling and storage pro%lded
9. — Storage stability data available reflecting storage of tissue, milk and egg samples pnor to residue
analysis (may be referenced to separate study)
Cntena marked with a ‘ate supplemental and may not be required for every study.
C.382
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Subdivision 0
Guideline Ret. No. Ill- 4 4j)
December 24, 1989
17l ) Mns M Po ftsy, -F g
GUIDAN( FOR SUMMARIZING ST JDI
Items in sum Iy sboe d IndDde the lte d ve 1 m C apset 2 of Ihi pmcbge and the specific items
listed below.
t. Compound(s) ted to aitlmal and comparison to residues on feed items.
2. Dosages or feeding Levels mtned and comparison to expected dietary burdens with brief explanation
of calculation of Latter.
3. Number of animals In each dosage group or feeding level.
.t Duration of dosing and tune at which residues were observed to plateau in milk or ew.
5. Method of sacrifice and time in hours from final dose.
6.7 Brief description of analytical method used to measure residues Including validation/recovery data and
statement of detection limits for tissues, milk andlor egp. Summarize the observed residues as a
function of feeding level in t*b The rnilkiegg table should show the residues as a function of time
so the point at which residues plateaued can be determined. For tissues, time is usually riot a factor as
sacrifice (TIOSI often occurs within 24 hours of the anal dose. However, if some animals are allowed a
k’ng.1 withdrawal period. these should be shown separately on the table. Unless une value LS
noticeably different than others in the same set, it would generally be adequate to provide the range of
‘.alues observed tar each tissue (or each feeding leveL Control values (i.e.. samples trom animals not
ingesting the ‘,csticide) should also be indicated. (See Tables Ill and IV for examples).
8. Brief description of dosing procedure. handling and diet of animals. average feed consumption (for each
feeding level), and coliecuon of ussue/milkicgg samples. (Storage addressed in item 91
9 Storage conditions and duration for tissues, milk and eggs between sample collection and residue
analysis. Summary of data showing stabthty of residues under iiniI r conditions (may be referenced to
separate study).
10 Identiflestlos of the tolerance petition number, if the study has been used to support a tolerance
C.383
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Table 111. Ssmpk summary table for feeding
stud ies - To c residue in tissues
Subdiv sLon ‘
Guideline Ret. No. 171.
December 24, 1
Feeding
PPM
JCIdnev
Total To c
—
Residue In Tlssu ,
Musc L. ._Fat —
Levej
0
ppm
5
<0.05
<0.05
<0.05
<0.05
ppm
15
0.15.0.37
<0.05
<0.05
<0.05
ppm
50 ppm
0.51 -0.97
1.37-1.93
<0.05 -4109
0.24-0.33
<0.05
<0.05
<0.05.0.12
0.14-0.47
Table
IV.
Sample
studies —
summaiy table
To c residues
for
in
feeding
milk
PPM Total Toxic Residue in Milk
5oom in d IS corn in dIet 50 corn in dj
•1 <0.01 <0.01 <0.01
1 <0.01 <0.01 0.03-0.07
3 <0.01 <0.01.0.04 0.21.0.37
7 <0.01 0.01.0.05 0.19.0.38
10 <0.01.0.02 0.02.0.08 0.25.0.43
14 0.01 0.03-0.07 fl-0.35
28 0.01 0.02.0.08 ‘S -0.41
Day before doelng Initiated ( ntrol samples).
c384
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Subdivision 0
Guideline Ref. No. l71 4(j)
December 24, 1989
17 1- 4 (J) Mat, Milk, Poultry, Egg—Dermal Treaiaeat
AQ ztAN(I ( iTER1A
These re ecnon criteria address studies in which the animals have been treated dermally with the
pesticide (as opposed to oral ingestion).
Does ur sI , the fotlowthg a pesace afteria?
L — Animals treated ac rding to label directions leading £0 m rmum residues
_ Maximum spray or dip concentration
_ Maximum spray volume per animal or maximum time in dip tank
L ._ Maximum quantity pour-on or dust formulation per animal
f_ Free access given to backrubbers and dust ba
— Minimum retreatment intervals
Maximum number applications data showing that milk or egg residues decline to
non-detectable levels within the minimum retreatment Lnterval specified on product
labels
f Registrant should write ‘NA’ (not applicabLe) if this type of application is not registered.
2. — Each type of formulation and application technique tested (although dips and high pressure
wetting sprays cover dusts and pour-otis, provided approximalely the same amounts of ictwe
ingredient are applied per animal)
3 — Multiple animals included in each treatment group to show variation in resid’ es (preferably at
least 3 for cattle and 10 for poultry)
4. Anima sacrificed within I to 3 days of 5nal application
5 — Total toxic residue m ured in edible tissues (muscle, fat. liver, cattle kidney) and milk or eggs
of both control and treated anii al5 using validated analytical method (description of latter given
or referenced)
6. Analytical method su ciently sensitive (0.01-0.05 ppm)
7 Milk or egp collected and analyzed daily until residues plateaued
3. Di.is4pLM)n of kindling and pesucidat treatment of animals and sample collection, handling and
s —
9 — Storage stability data available reflecting storage of tissue, milk and egg samples prior to residue
analysis (may be referenced to separate study)
Cr tena marked with a s are supplemental and may not be r uired for evety study.
C-385
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Subdivision
Guideline Ref. No. 17
December 24,
17 1 .4 (J) M i, Milk, Poultry. Eg-Damal Treatment
GU1DAN( FOR SU! Q (AR 1G STUDIES
ftec in summary s uld indudc the 1t d cmaed in C ap(ct 2 of tb p ge and the spet & it
Listed below.
1. Comparison of application rates. retreatment intervals, and number of applications in study to those
permuted on product labeLs. If mammum number of applications not examined, data showing level of
milk or egg residues after minm um retreatment interval has elapsed.
2. Formulations and application techniques tested.
3. Number of animals in each treatment group.
4. Method of sacrifice and time elapsed from final dose.
5.7. BneI descrtpnon of analytical method used to measure residues including validationirecovery da*a and
statement of detection limits ftr tissues. milk and/or eggs. Residues observed in edible tissues and milk
or eggs. If different formulations and/or application techniques were examined, tabulate the results with
a separate column for each formulation and/or application technique. The milk/egg table should show
he residues as a function of time so the point at which residues plateaned can be determined. Also. if
animals are sacrificed after various withdrawal periods, residues in tissues should be shown as a
function of time. Unless one residue value is noticeably different than others in the s zne set. it wou
generally be adequate to provide the range of values observed for each tissue for each type of
treatment. Control values (i.e.. samples from animals not treated with the pesticide) hould 4150 be
indicated.
8 Bnef description of pestiadal treatment. handling and diet of animals, average feed consumption. and
collection of ussue/milklegg samples. (Storage addressed in item I
9 Storage conditiou and duration for tissues, milk and eggs between sample collection and residue
analysis. Summary of data s ing stability of residues under similar conditions (may be referenced to
separate study).
10. Identification of the tolerance petition number, if the study has been used to support a tolerance.
C .386
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Subdivision 0
Guideline Ref. No. 171-4(k)
December 24, 1989
171 -4(k) Msgult o( the Residue — Crop FIeld Trta
AccEPTANcE CRf ERIA
Does u study meet the bUo tng a ptaaee cruczia?
1 — Crop treated according to label directions leading to ma mum residues:
— Ma mum application rate and number of applications
— Minimum retreatment intervals
— Minimum prebarvest interval
— Minimum spray volume
— Data reflects modes of application on label: ground. aenal, oil dlluenLs, use of adjuvants such as
surfactants and spreader/suckers
3 — Representative formulations examined
4 — Trial locations represent aU principal growing regions (adequate ‘geographic representation’)
Q minor crop for which regional registration is accepted
5 — Sufficient number of tnal conducted to determine an appropriate tolerance or ma omum residue
level (factors to consider include seasonal and yearly variations, differences in varieties and
cultural practices, the importance of the crop. the concfty of the pesticide and its a iftv to
tranciucate. and the type of usc-e.g., early seasonipre.cmergence versus late season fo iar)
— Total to c residue measured by validated analytical method (description provided or referenced)
on all parts of crop imed for food or feed (except for feed tiems which are restricted by product
labels)
7. — Description of sample treatment, collection, handling (including any washing or trimming) and
storage provided (covering periods trout pLanting to harvest and from harvest through analysis)
8. Dates of Irea* 1, harvest, enny into storage, and residue analysis
9 — Storage stability data available reflecting storage of crop samples (may be referenced to separate
s)
JOTE: P W”- should reftr to the document ‘GUIDANCE FOR PHASE 2 OF REREGISTRATION
FOR PESTICIDE MINOR USES AND LOW VOLUME PESTICIDES’ to determine whether all of these
data requiremenD apply to minor i es.
Criteria marked with a • are supplemental and may not be required for every study.
0.387
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Subdw isu
Guideline Ref No. 17t-
December 24. 1989
171-4(k) Ma Atede ci the Residue - Qop Flee Trin
GU1DAN W R SUP.Q4ARIZINO STUDIES
Iteum in summary should l ude the Iteum dis am1 In C apter 2 ci this pa ge and the sp ñr ftc
listed below.
I Comparison of application rates, uminp, and spray volumes in field trials to those permitted on
product labels.
2. ComparIson of application modes and spray additives in field trials to those permitted ott product
labels.
3 Formulations tested and ma.idmum residues found for each one.
4-S Brief discussion of how trials provide adequate ‘geographic representation’ for the crop and sufficient
data to determine the m imum residue level expected from actual use of the pesticide.
6.9 Brief description of analytical method used to me ure residues including validation/recovery data (latter
may be haled in a footnote to the s ln.ry table). Stunmarize the results of field trials for each crop
in a Al a minimum, the table should include information on location of each trial, the rate and
number of applications, the type of application eqwpmenl (i.e., ground or aerial), the spray ‘.olumes
(GPAagallofls per acre), the dates of applications (or intervals in days between apphcauonsl. the ttm
In days from the final application to harvest/sampling, and the Level of the residue of concern.
If different portions of the residue are determined by separate analytical methods, the level of each
portion should be listed along with the total residue. When more than one sample having the same
location, rate and prehazvesl Interval has been analyzed, the indMdual values should be showli rather
than a mean. Residues on treated samples should not be corrected (or controls. Correction for
recoveries is not desirable, but if it has been done should be clearly indicated. Each crop pan on
which a tolerance is to be set (sec Table if of Residue Chemistry GuideUnes-$ubdivzsion 0) should be
included within the table QL a separate table may be generated for each crop part (e.g.. corn forage.
fodder, and grain). Any postharvest treatment of the samples such as washing or trimming should be
noted. If different formulatlone or modes of application (e.g., pre.emergence venus post -E) result in
significantly different residue., a separate table is advtsable (or each.
Applicatlas ma. and tlMMgI other than the expected use should be included in the table to address
issues su c seed processing studies. However, registrants should use judgment in cases
where a pupurlios c i the data is not representative of the proposed use. The usefulness of the
summary be lees If eesiva maminallon of it U ne esu1y to determine which data are
applicable $ as g wlar or estimating dietary exposure. As a related comment, data that
obviously do not meet the resection criteria should not be induded in the table unless a written
justification is provided.
Other information that needa to accompany this portion of the crop field trial summary includes
residue vaIuc on untreated (control) samples and sample storage conditions and duration (and how the
latter compare to controlled storage stability studies). As it may be dlMculi to put all this information
C .3
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Subdivision 0
Guideline Ref. No. 1714(k)
December 24, 1989
plus that mentioned above in the table Itself. it would be a ptable (ft m t ‘° provide the
range$ of control values, storage periods, etc. from the field triaLs within the brief narrative summary
preceding the table or as footnotes to the table. If trials are run in more than one year, the year
should be indicated (or each triaL When the field trials for one crop are located In more than one
MRJD# (or Accession 0), the MRID# number for each trial should be shown within this section of
the summaty. A mpany study number, if assigned, would also be useful 33 it would allow more rapid
location of the raw data when the latter need to be examinet
(See Table V (or a case where all the studies tall under one MRID#).
10 identification of the tolerance petition number, if the study has been used to support a tolerance
C.389
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Subdlv slon ‘
C,wdellne Ret. No. 171-
December 24,
Table V. Sample summary table rot crop geLd
- Residue of parent pesticide plus
rnetabolite on lettuce
0 & Rate Equip Dates of PPM PPM PPM
L tion of Aenin & OPA Anoin ffl tareni Merab To l
Town 1, CA 3 0.10 0 30 2/5, 2119, 3/10 7 1.50 0.27 1 77
14 0.82 047 1.29
Town 2. CA 3 0.08 A 8 3/8, 3,21, 417 15 0.52 0.21 0.73
Town, NY 2 0.10 0 25 5 (2 ., 5,23 8 2.47 0 17 2.64
4 1.34 035 1.69
Town, Co 3 x 0.10 0 35 6 /9. 6,21, 713 4 0 ,28 0.30 0.58
Town. AZ 2 x 0.10 A 5 10/12, 10128 13 037 0.08 0.45
W’hole, unwashed lettuce heads were analyzed. Only obviously decomposed wrapper leaves were
removed pnor to analysis.
Application rate in pounds active ingredient per acre
G ground application, A—aerial. OPAugallons per acre
PHI — preharvest interval in days
Control samples contained <0.05.0.08 ppm apparent residues of parent and <005 ppm
metabolite.
Lettuce sa ’nples (torn above trtals were stored at -2Q C (or 2-6 months prior to residue analysis.
Storage staothcy studies using apples. wheat and lettuce showed no loss of residues after 12
months at .20 C (Reference. MRID#).
Recoveries from control lettuce samples spiked with 0.1 ppm parent and analyzed concurrently
with the field treated samples were 78-103%. Recoveries of metabolite (0.05 ppm) were 72.
97%. (If method b been submitted separately, provide MRID# or other appropriate
reference.)
C.390
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SubdivisiOn 0
Guideline Ret. No. 17 1 .4 (l)
December 24, 1989
17 1 -4 (1 ) Proi .r ed Fo /F
AQ rTAN fl’ER1A
Does ur uody nu tM btIo thg punce altesia?
— The raw agricultural commodity (r.a.c.) sampLes that re processed contained field-created
detectable residues (preferably at or above the tolerance) Q r.a.c. was treated in the field at
exaggerated rates in an attempt to get detectable residues Q , spiked r.a.c.s were used with
data available to show that r .a.c. residues st solely on the surface
2. — The r.a.c. was processed using procedures simulating commercial practices
3. — The total to c residue was measured with a validated analytical method (description provided or
referenced) in the r.a.c. and all us byproducts listed in Table 2 of the Residue Chemisny
Guidelines
4 — Descnpuon of pesticidat treatment and processing of r.a.c. and collection, handling and storage
of samples provided
5 Storage stability data available reflecting storage of r.a.c. and byproduct samples (may be
referenced to separate study)
Criteria marked with a • are supplemental and may not be required r every study.
C . .391
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SubdMMo
Guideline Ref. No. l7
December 24, o’
171-4(1) ProoraiW Food/Feed
GUIDANCE )R SUt *.4AR iO Sfl.IDIES
Items in summary should Indode the lt 4’4 in C apter 2 th p r and the spe flc nc
(istut below.
1. PPM pesticide on raw agricultural commodity (r.a.c.). Description of procedure used o deposit
pesticide residues on r.a.c. For field treated r.a.c.’s. description of mode, rate, and timing of pesticide
application. For fortified sampLes, description of fortification procedure and summaly of data showing
r.a.c. residues st soLely on sur of crop.
2. Brief description of processing techniques with comparison to those employed commercially.
3. Brief description of analytical method used to measure residues including validauonirecovery data.
Levels of total toxic residue in the ra.c. and by-products (including control samples) Listed in TabLe II
of Residue Chemisuy Guidellnes
4 Storage condiuons and duration for r.a.c. and byproducts between processing and residue analysis.
[ Description of pesticide! treatment and processing covered by items 1 & 2.
5. Summary of data showing stability of residues under storage conditions described in item 4 (may be
referenced to separate study).
6. Identification of the tolerance petition number, if the study has been used to support a tolerance,
C .392
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Subdivision 0
Guideline Ref. No. 171 .5
December 24, 1989
171-5 Redu ion of Resld
A PTAN ITER1A
Does wur study e* the followthg a pua fteria7
A. DEGRADATION STTJDIES
1 Description of sample history provided for the raw agricultural commodity (r a_c.) This could
include items such as the pesticidal treatment (application rate, preharvest interval, where grown.
etc.). date and location of purchase . and storage condiuons from harvest or purchase until
processtnglwash ing!Cooking
2. — Detailed description of residue reduction procedure used on r.a.c. Such procedures may include
trimming, washing. peeling, cooking, or storage of the commodity under simulated commercial
or home food preparation conditions.
3. — Descnption of handling and storage of samples from time of residue reduction procedure untLi
analysis for residues
4. Total toxic residue measured with a validated analytical method (description provided or
referenced) in the r.a.c. both before and after the residue reduction procedure
5 — Storage stability data available reflecting storage of r.a.c. and ‘processed’ samples from time of
residue reduction pro ure until analysis for residues
B. MONITORING STUDIES
1. — Descnpuon of sample history provided for the commodity being monitored. This could include
items such as sampling Location (state, city, supermarket - large chain or local store. etc.).
brand (national vs.local), fOod form (e.g. frozen, canned or fresh peas), date and location of
purchase, and storage and handling conditions from purchase until residue analysts
2. — Detailed description of the statistical design of the study. This should include discussions of
reprcscntauvenesa of sampling (does the sampling provide adequate representation in terms of
a,mmodiII , loul on, brands, types of procassing, etc.), and aocuracy and precision of results
3. — Total t c residue measured with a validated analytical method (description provided or
r zite4 ) In the mmodi*y
4. — Storage stability data available reflecting storage of samples from the time of sampling until
residue analysis
Criteria marked with a are supplemental and may not be required for every stu4
C-393
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Subdivi
Guideline Ref. Mo.
December 24, 1989
171.5 8d 1n ______
GUIDAN( )R SUMMAR1W ’G STUDIES
Items in sum iy shoeld Iadnde the 1 d ied I a C apta 2 th ge sad the sp & items
Usted bekiw.
A. DEGRADATION STUDIES
As noted on the prevlos page, the orpnhzatlon and ntent of the summ iy should follow the
rejection cntena for the study.
I. Descnption of sample history for raw s icuItural ammodny.
2. Description of residue reduction procedure used on raw agricultural ummodity.
3. Conditions and duration of storage of samples from time of residue reduction until analysis of residues.
4 Brief dcsa’lption of anaMI l method used to measure residues including vulldation/rectwery data.
Levels of total toxic residue in commodity both before and after residue reduction procedure.
5. Summary of data showing stability of residues in samples stored under sumlar conditions to those note?
in item #3 (may be referenced to separate study).
6. identificution of the tolerance petition number, If the study has been used to support 3 tolerance.
B. MONITORING STUDIES
As noted on page 1, the org”n4 tion and content of the snmmary should follow the rejection cntena
for the study.
I. Description of sample hktoey 3r monitored commodity.
2. Description of the stadadeni dlgn of the study.
3. Brief desaiption of snalydal method med to measure residues Including validadon/recovery data.
4. Summiry a abowing sbilhsy of residues in samples stored from purchase until residue analysis
(may be i- -1 to separate study).
5. Identi cetios the rolmeuee petition number. if the study has been used to support a tolerance.
C-394
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Subdivision 0
Guideline Ref No.
December 24, 1989
171-6 Pm xiaed ToIerua
ACCEPTANCE CRflERIA
Do vur study m t the *Uow ng pcan aitena?
Tolerance expression includes complete, accepted chemical names of parent pesticide and
metabolices of toxicological concern
2. — Tolerance includes proper name or description of commodity or crop group
3. — Tolerance high enough to cover any residue values reasonably expected based on residue data
(i.e., tolerance not based on average residues, but on maximum residue)
4 — Tolerance not set higher than necessaty unless inordinate multiplicity of tolerance levels needs
to be avoided
5 — Specification of conditions of use of pesticide for food additive regulations covering use in food
handling establishments
Criteria marked with a e are supplemental and may not be required for every study.
C-395
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Subdivision
Guideline Ref. No. l
December 24, 1.
171 -6 Propo.cd ToIerani
GUIDANCE FOR SUMMARIZiNG SflJDIES
IteuL5 in snin ry 3he ld i ade the ke d ed in ( apIe 2 ot thk p g and the spook itcn
Listed below.
? 4o summaiy is required kiT this guideline requirement
C-396
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Subdivision 0
Guideline Ref. No. 171-7
December 24, 1989
171-7 R $ouabIe Grouw In Suppon of Petition
ACCEPFM4c CRrr u ,
Does ,ouz study mcet tbe 1kJu4ng a pIan aiterIn?
1. Rationale of how residue data support tolerance
2. Explanation of any aberrant residue values
3 — Explanation for omission or Substitution of requirements set forth in Residue Chemistry
Guidelines.
4 Brief discussion on sensitivity of analytical method and its ability to measure total toxic residue
Criteria marked with a • axe supplemental and may not be required for evety study.
C-397
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Subdiv i io
Guideline Ret. No.
December 24, t,,.
111-7 R o Mr Gtouath in Suppi t 01 P don
OUTDANC FOR SU!IO 4ARJ22NG S’TUDtES
Items iA sn ” Iy should (he Ltams di.cuued Ln c apser 201 thh p r gc and the sp items
t ted bdo .
No summary is required for this guideLine requirement.
C .398
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Subdivision 0
Guideline Ret. No. t7l t3
December 24, 1989
171-U Submiual of Ana1yti I Reter Slandaids
AcL U’TANcE CR ER1A
Does y° Suidy the ft lk uing ePta alteria?
1. — 2 grams each of purified analytical standards provided for pesticide and principal degradation
products or meubolites
2. — Reference standards sent to proper address:
Pesticide and Industrial Chemicals Repository (MD8)
U.S. Environmental Protection Agency
Research Triangle Park. North Carolina 27711
3 — Material Safety Data Sheet included as required by OSHA in 29 CFR 1910 1200
4. — Letter of transmittal accompanying standards
— Purity of standards
— Analytical methods used to assay standards
— Statement of principal impurities
— Punflcauon procedures
— Storage requirements
— Special precautions for safe handling
Criteria marked with a ‘ are supplemental and may not be required for every study.
C-399
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Subdivj i 0
Guideline Ref. No. 17
December 24, L
171-13 Sub fttaI of AnaJ tJ j Standarc
OUIDAN( R SU?.O,4ARJ O SflJDEES
fte La summaiy sbouid lndi e the ft d1S(H 1 L a ( apI 2 of th e and the spod& ite
Listed below.
1. Description of sample submitted. including:
Name of pesticide and pnnczpal degradation products or metabolges
Punty
• Analyti l methodolo r used
• Statement of pnncipal impurities
• Purification procedures
• Storage requirements
• Special procedures for safe handling
C-400
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SUBDIVISION R
201.1 Spray DdfI Droplei Size Spectrum . . 402
202.1 Spray Drift Field Evaluation .
C-40 1
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Subdivision R
Guideline Ref. ‘lo. 201
December 24, 19
1-l Sprsy DrIft Droplet SL Spe ii
AQ TAN cRrrERIA
Does your snMIy — - tke MIou4ag pta aIterle?
1. — The test chemical used was either the proposed formulated end.use product or of the same
formulation category as the end-use product to be registered.
2. — The percentage or degree of purity of the at was reportect
3 — The application rate was reported.
4. — The physical properties of the in-tank formulation mix were gwen including surface tension,
viscosity, density, vapor pressure. etc
5. — The collection surfaces or devices were identified along with the equipment limitations
(including the resolution of the microscopes. particle measuring systems, or air samplers. and
the spread factor of the droplets on the various collection surfaces (cards) for droplet sizing).
6. — The test site was specified (e.g., wind tunnel or field evaluation).
7. — The environmental conditions that prevailed during the test (temperature. relative humidity,
air velocity) were provided.
8. The noules used were those most likely to be used and cause drift (or the application of
that pesticide or the intended use pattern.
9 — The nozzle pressure, flow rate, and orientation to the airstream were reported.
U). — The particle size distribution versus both cumulative percent volume and droplet number
(frequency) were reported.
Criteria marked with a are uipplemenial and may not be required for evety study.
C-402
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Subdivision P.
Guideline Ref. No. 201-1
December 24, 1989
)I-1 Spray Drift Droplet Size Sp rrusn
GUIDANCE FOR SUMMARIZING STUDIES
Iteu s in snmmaIy should indale the iie s dh ed In ( aprer 2 of this pa age and the specific ite
listed below.
A Summ2ly of Condusio
B. Test Material
1 Test material used (formulation, etc.)
2. The punty of the test substance and pounds A! per acre applied
3. Physical properties of the in-tank formulation rmux.
C. Test Methodolo*y
1. Application rate/method
2. Full description at’ collecting surfaces/devices including limitations of such surfaces/devices
3. Identification and description of test site
4 Description of environmental conditions prevalent during testing (e.g. temperature. relative
humidity. air velocity)
5 Nozzle type, size, pressure, flow rate and orientation to air stream and justification for USC
D. Reported Reiults and Dic i .
1.The particle size distribution venus cumulative percent volume (tabular rmat).
2. The particle size distribution venus dropLet number (frequency distribution).
3 The results tabulated to indk te a 10, 50, and 90th percentile of particle size distnbuuon with
respect to droplet vlumc and number (tabular format).
C-403
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Subdivi i R
Guideline Ref. No. 202.1
December 24, 1989
3fl-1 Spray DrilL Add Evai rin.
A PTANcE *ZRXTERI
i ) UT - t e aowthg aitcr&
1. The teat chemical used was either the proposed formulated end-use product or of the same
formulation categoiy as the end-use product to be registered.
2. The test site specified (e.g. open field, cropped field, forest, grove, etc).
3. The envtronmencaj ndkions which prevailed during the test (temperature at two levels.
relative humidity, air velocity and direction, rainfall or watering regime (overhead imgauon
systems)J were provided.
4. A diagram of the plot or area was included in the report which Indicates north, swath width,
and onenlauon, prevailing wind direction, and location of the collections stations.
5. The type of application equipment used was reported.
6. The nouies and associated equipment were those most Likely to be used for the application
of that pesticide or the intended use pattern.
7 The estimated minimum and maximum nozzle-to-target height was reported.
8. The application rate in quantity per unit area (of plant or land Surface) or in quantity per
minute flow rate (gailons.per.mmute or Liters-per-minute) was reported.
9. Diluent., or camera as weLl as the extent of dilution were identified along with possible
adjuvants, tank mixtures with other pesticides, and total spray volume.
10. The physical properties of the in-tank formulation mix were given including surface tension.
viscosity, density, and vapor pressure.
11. — Controls were used to note any possible decay or other loss of pestiude during the
collection, transport, storage, and analysis.
1 . The distribution of the collection surfaces were of sufficient number to e.stablish a definitive
uninterrupted picture of deposits across the treated swath as well as Outside the target area.
13. The placement of the collection surfa (either at the soil sur ce or at the height of the
surrounding canopy) was reported.
14. — If air samplers were emplo d, were they at least three downwind locations (one at the
farthest downwind she preferable).
15. The quanutlee of pes’ t ’lile at each collection station were reported.
16. The stability ratio (Bared preferred) was calculated.
Criteria marked with a • are Supplemental and may not be required for every study.
C-404
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Subdivi i R
Guideline Ref. No. 202-1
December 24, 1989
Y2-1 Spray Drift Field EvaluatioQ
GUIDANCE FOR SUMMARiZflqo S’flJDLE
Items in snmm ry should include the items discussc J in Chapter 2 of this package and the specific items
listed below.
A. Summzny of ndtisioi
B. Test Material
1. Test material used (formulation, etc.)
2. Percent Al in test material
3. Diluents, carriers, adjuvants contained in tank mixture
4 Physical properties of the in-tank formulation mixture (e.g. density, surface tension, viscosity,
vapor pressure etc)
C. Test Methodolo
1. Application rate,’method
2. Full description of collecting surfaces/devices including limitations of such surtaccs/devices and
distribution of devices within the test site
3. Identification and description of test site (diagram).
4. Location of air samplers if used
5 Description of environmental conditions prevalent during testing (e.g. temperature, relative
humidity, air velocity type, Barad stability ratio).
6. Nozzle type, size, pressure, flow rate and orientation to air stream, and justification for use
7. The speed, height, and type of the application equipment over the ground
8. The eadmated no 1e-to.larget height
C -405
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Subdivj 10 R
Guideline Ref. No. 202.1
December 24, 1989
D. Reported Rule
1. The particle size distribution versus cumulative percent volume and versus droplet number
(frequency) if the droplet size spectrum study was performed as part of the field evaluation
(tabular format).
2. The celculated Barad stability ratio (tabular format).
3. The quanüti of pesticide at each collection station and the intended quantity of pesticide per
area (g/hectare) (tabular format).
4. The extent of downwind spray drift evaluated with respect to possible exposure to the non-target
organism(s) that may be detrimentally effected (tabular format)
E. Dua&isjoo of result
I. Exposure assessment of spray drift to non-target organisms.
C-4c
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SUBDIVISION U
230-236 Appll tor Exposure Monitoring: Passive Dosimet . . . 408
C-407
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Subdwtsion U
Ouidelrne Ref No Z3O .2
December 24. l98
Act ZTANcE fl 1A
Dam ur stedy meet t bLk w(ng psan aitefla?
1. — Typical enduse product of the active ingredient tented.
2. — End.use product handled and applied using recommended equipment. application rates, and
typical work practi .
3. — Exposure monitoring using at least five replieaces at each of at least three sties for each job
function with the exception of pilots. Pilots should have at Least three replications at each of
at least three sites.
4 — Monitoring period is sufficent to collect measurable residues but not excessive so that residue
loss o urs.
5 — Dertnal and/or inh t2ÜOfl exposure monitored by validated methodologies. Biological
morutoring is consistent with and supported by pharmacokinetic data accepted by the Agency.
6. — Quantity of active ingredient handled and duration of monitoring period reported for each
replication.
7 — Clothing worn by each study participant and location of dosimeters reported.
8. — Quantitative level of detection is at least 1 hg/c&.
9 — Storage of samples consistent with storage stability data.
10 Efficiency of extraction in laboratory provided as mean plus or minus one standard de iaUon
Lower 95’T confidence limit is not less than 70% based on a minimum of seven repIiL3ttons
per fortification level or prior Agency approval of extraction methodolo ’ pro idcd
t._ For each matrix at least one field lomfication sample per monitoring period per forti1ic iion
level. At least one fleld blank per worker per monitoring period (or each maIrL .
L2._ When collecting urine for biological monitonng. collection should involve 24 hour urine
samples. A minimum of one baseline, pre.aposure 24 hour sample must be collected 24
hour samples must be collected for the day of application and for sufficient days
posupplicanon as determined by the excretion profile of the pesticide.
Criteria marked with a’ are supplemental and may not be required for every study.
C.408
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Subdivision U
Guideline Ref. No. 230-236
December 24. 1989
230-Z36 Appli tfon Expa6uzc Monitonng Passive D imetry
GUIDANCE FOR SUMMARI7 4G STUDIES
Items in summ2ly should indude the items diiaissed in Cbapter2 of th package and the specific items
listed below.
1. Form of pesucide tested.
2. Product handled and applied.
a. equipment
b. application rates
c. total amount of material handled for each activity
d. duration of monitoring periods
3 Pilots
a. Number of sites monitored
b. Number of replicates per site
4 Dermal and/or inhalation exposure and biological monitoring methods
5 Clothing scenario studied for each study participant including location of dosimeters and comparison
to label directionc concerning clothing.
6 QAiQC data including quantitative level of detection, method efficiency, storage stability, (ic!d
recovery for each matrix analyzed.
7 Number of field samples and blanks per worker per monitoring period.
8. Sample schedule for urine sample collection for biological monitoring.
9. Description of compound metabolism for biological monitoring.
10. Dermal and inhalation reported for each replicate and entire population as milligrams of exposure
per pound of active ingredient handled (mg/lb/al) and on a per hour basis; raw data should be
reported as as that corrected for field recovery (less than 100%).
C-409
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C-410
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
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Dec mbcr 24. 1989
PHASE 3 TECHNICAL GUIDANCE
APPEND(X D. OVERVTEW DOCUMENT FOR SUBDIVISION F
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December 24, 1989
PHASE 3 TECHNIcAL GUIDANCE
OVERVIEW OF SUBDIVISION F
HAZARD EVALUATION: HUMAN AND DOMEsflC ANIMALS
This document provides an overview of the technical guidance for Subdivisi F. Table D-1 lists
EPA publications pertaining to Subdivision F that are available through the National Technical
Information Service (NTIS). The first document on the list, the Pesticide Assessment Guidelines (PAG,
sometimes referred to simply as ‘The Subdivision’), provides general guidance. It is supplemented by
the Data Reporting Guidelines (DRG) published as a senes of addenda. Table D . .2 shows where to
locate particular topu in the general guidance and Addenda. The body of the document reviews major
changes since the guidance was onginally published in 1982; clarifies some apparent inconsistencies
between the Pesticide Assessment Guidelines, Data Reporting Guidelmes, and Standard Evaluation
Procedures; provides some new guidance; and indicates future directions that the Agency is considering.
The document is organized into parts to address the following categones of toxicity studies:
I Acute Toxicity Studies
II. Subchronic Studies
III. Chronic and Long Term Studies
IV. Teratology Studies
V. Reproductive and Fertility Effects Studies
VI. Neurotoxicity S’udies
VII. Mutagenicity SitiJies
VIII. Metabolism Studies
IX. Dermal Absorption Studies
D.1
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December 24.
PHASE 3 TECHNiCAL C%JtDAN(
Table D-l. EPA GUWANCE DOCUMENTS FOR SUBDII’7SJON F
NTIS Number EPA Number
PESTiCIDE ASSESS) 4T GU1D 24ES (FAG)
Humans and DomefliC An1ffiaL (revised) P386.108958 540,09.84-014
DATA REPORTENG GU N (DROYADDENDA
Humans & Domesuc AnimaIs-MdendUm 1 P886-248184 540i09 .86 -150
Humans & Domestic Mtimal$-MdCfldum 2 P388-162292 540 109 -88 -023
Humans & Domestic M n I -M4efldUm 3 P388-161179 540i09.88-024
Humans & Domestic AnimaIs-Mdefldwn 4 P388462227 540 V9 -88-028
Humans & Domestic Anunals-AddeTidUIn 5 PB88- 162219 540 ,09-88.029
Humans & Domestic Anim tI -Addefldnm 6 PB89 - 1240T7 540,09-89 . 001
Humans & Domestic MIm k.AddendUflt 7 P389424085 540 09-89 .008
Humans & Domestic Anunab-Addendunt 8 P890422221 540,09.90-071
STANDARD EVALUATEON PRO UR (SE?)
Guidance for Evaluation of Eye Irritation Testing P889 .124572 540d 09 -8 8 -lO 5
Inhalation Toxicity Testing P889.100366 540,09-88-101
OncogenicIty Potential: Lafl8 Term Rodent Studies P886-129400 540 09- 5-019
Teratolo ’ Studies P886-129392 540 ,09.85.018
Toxicity Potential: Subchronic and Chronic Exposure Studies P886-129418 540,09.R5 -020
OTHER GUIDANCE
Position Document: Maxunum Tolerated Dose P888- 116736 540.09-8 -003
Re .ised Policy foi Acute To dty Testing PB9 O- 122151 540/099th072
D-2
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
Thbk 0-2. WHERE TO FIND INFORMATION IN SUBDIVISION F GUIDANCE DOCUMENTS
TOPIC LOCATION
___________________________________________________________ Page in PAG Addendum Numi
Organization and Philosophy of Subdivision F I
II. The Standards for Acceptable Testing 3
111 Discussion of Individual Tests U
80 OVERVIEW, DEF1NrrIONS, AND GENERAL REQUIREMENTS 20
80-I Overview 20
80-2 Definitions zo
8( 1-3 General Provisions 21
80-4 Reporting of Data 28 1-8
80-5 Combined Testing 32
81 ACUTE TOXICITY AND IRR iTATiON STUDIES 34
81- 1 Acute Oral Toxicity Study 34 8
81-2 Acute Dermal Toxicity Study 39 8
81-3 Acute Inhalation Toidcity Study 44 6
81-4 Primary Eye irritation Study 51 2
8 1-5 Primary Dermal Imiation Study 55 3
81-6 Dermal Sensitization Study 59 4
81-7 Acute Delayed Neurotoxicity of Organophosphorus Substances 62 5
82 SUBCHRONIC TESTING 66
82-I Subchronic Oral Toxicity (Rodent and Non-rodent):90-lDay Study 66 8
32-2 Repeated Dose Dermal Toxicity: 21-Day Study 76 8
82-3 Subchrontc Dermal Toxicitr 90-Day Study 83 8
82-4 Subchrontc Inhalation Toxicity 90-Day Study 92 6
82-5 Subchronic Neurotoncity 90-Day Study 103 5
83 CHRONIC AND LONG TERM STUDIES 107
83-1 Chronic Toxicity StudIes 107 8
83-2 Oncogerncity Study 117 8
83-3 Teratogenicity Study 126 1
83-4 Reproductive and Fertility Effects 130 8
83-S Combined Chronic Toztdty/Oncogenicity Studies 137 8
84 MUTAGENICITY 147
84-1 Purpose sad General Recommendations for Mutagenicity Testing 147 8
84-2 Mutndthy Tests 148 8
85 SPECIAL STUDIES 152
85-1 MetabolIsm Study 152 7
85-2 Domestic Animal Safety Testing 156 (18)
85-3 Dermal Absorption Studies of Pesitcides (Reserved) (19)
D -3
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December 24, t989
PRASE 3 TECHNICAL GUIDAr4C!
I. ACUTE ,aCrrY SIUDIES
Guidelines 81-1. 81-2. 81.3 (for oral. dermal and inhalation toxicity) have been changed since 1982
to reflect the Agencys objective to reduce the number of animals used to evaluate acute toxicity while
providing adequate information on chemical safety. A new Section (4) recommends the following:
1. use of existing data on structurally related chemicals
2. use of limit test
3. estimation of lethal dose
4. multipLe endpoint evaluation
In 1984, Section (g) was changed to ‘A concurrent untreated control is not necessary. A vehicle
control group should be run concurrently except when histoncal data are available to determine the
acute toxicity of the vetucle. ’ A proposal is under consideration to remove this requirement and not
require arty controls (as was the case in 1982).
The following changes refer to the Revised Policy for Acute Toxicity Testing’ issued on Sept. 22.
1988. and the proposed revisions to the acute oral. dermal and inhalation guidelines (Ut preparation)
which are based on this rcvised policy. In the revised policy there were major changes under the
headings ‘sex’ and ‘numbers’, as follows:
Sex: Where previous guidelines required equal numbers of animals of each sex for each dose level,
the 1988 revised policy calls for consideration to be given to Limiting studies to the more sensitive
sex, using previous history on the chemical class to make the determination, and testing of a few
animals of the other sex for confirmation.
Numbers: Where previous guidelines required at least 10 animals (5 of each sex) at each dose le ei.
the 1988 revised policy urged the use of abbreviated test methodologies, whenever possible
(approximate lethal dose, moving average method, up-and-down methods were suggested) which use
the lowest fea le number of animals.
The 1984 revisions removed from one part of the guidance (Guideline ) -4) the requirement for
measuring lethal dose within a 95% confidence interval of 20% of its median when technically feasible.
Through an editorial ovcrs lg*t, the requirement was still referenced under (i) ‘Data and reporting’ (3)
‘Test report’. Under the 1989 proposal, this mistake will be corrected and the requirement will no
longer appear in the revised guidelines.
a SUBO1RONIC SI1JD(ES
k Subcbm I ud’ ’ ( rodsal ned erodent) 90-day study ( -1).
The Agency h been lthrming registrants that one of the major purposes of the subchronic rodent
study is to determine the proper doses for the chronic/oncogeniClty study. We have also been informing
them that all data relating to estimating the maximum tolerated dose and presenting results of dose-
related effects should be presented in a clear manner.
D .4
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December 24. 1989
PHASE 3 TECHNICAL GUIDANCE
Purpose
The 90-day oral subchronic toxicity study in the rodent may also be used to estimate the lowest dose
level that is likely to induce life-threatening toxicity in the rodent chronic/oncogenicity feeding study.
This dose level is referred to as the Maximum Tolerated Dose (MTD). When using the data obtained
from the subchronic study (or determining dose levels to be used for a chronic/oncogenicity feeding
study, the MTD should be taken into account since it is desirable to determine the possible oncogenic
effects induced by the test substance at as high a dose level as possible. I I should be noted thai
esumatton of the MTD from the subchronic study may require testing more than 3 dose levels. For
further guidance on MiD ’s, a position paper on the selection of an MTD for oncogenicity studies
(Pesticide Programs) has been published through NTIS, and additional information is provided in the
Phase 3 Acceptance Criteria for Guidehue Studies 83-2 and 83 5. It is recommended that a 90-day
study be camed out on mice if a mouse oncogenicity study is planned.
Test prot dures
When determining dose levels to be used for the rodent chronic/oncogenicity study, the MTD (the
lowest dose level that is likely to induce life-threatening toxicity in a chronic/oncogenicity feeding study)
should be estimated. Estimation of the MTD front the subchronic Study may require testing more than
3 dose levels.
Data and reporting
• The hisIopatholo data should be summanzed in at least two ways:
- A summary individual animal data tableS for example, list individual animal number across the
top 01 the table versus indtvidua lesions by tissue (can be further broken down by grade within
the table; i.e., P=present, l=mtmmal, 2=slight, 3 moderate, A=autolysis , X=not remarkable,
(=in omplete section, etc.) down the side of the table.
• A summary lesion incidence table: for example, list dose group across the top of the table versus
individual lesions by tissue down the side of the table. It is important to include number
of tissues examined per dose level in this table.
• When considering body weight changes, include body weight gains in treated animals when compared
to controls (this is particularly helpful when estimating the MTD).
• Include food emciency lcuIations (body weight gains versus food consumption). This is also iaseM
when estimatIng the MTD.
B. Subchron 4IIk6I4n tlukfty 90-day study (82-4)
In specific cases the Agency has modified this requirement to a 21-day inhal2tion study. This
requirement should be discussed with the Agency prior to proceeding with the study.
D-5
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December 24, 198
PHASE 3 TECHNICAL GUIDANC1
ifi. G1RONIC AND LONG T M STUDIES
A C ronIc 1 d$y Study ( .1)
The Agency has had problems interpreting data presented by registrants, especially histopathoto t
data. Therefore we suggest the foLlowing:
HistopathoLo r data should be summarized in at least two ways:
• A summaiy indMdual animal data table: (or example, fist indtvidual animal number across the
top of the table versi individual lesions by tissue (can be further broken down by grade within
the tables I.e., P.presenl. l minimal, 2—slight, 3—moderate, A —aucolysis, X —not remarkable,
I=tncomplete section. etc.) down the side of the table.
• A summary lesion incidence table: for example. list dose group across the top of the table versus
individual lesions by tissue down the side of the table. It is important to include number
of tissues examined per dose level in this table.
• When considering body weight changes, include body weight gains in treated animals when compared
to controls.
• Include food efficiency calculations (body weight gains versus food consumption).
B. Oncogenicity study ( .2)
The following suppk!nentary guidance is provided:
• Data should be available to indicate that the animals were tested at a sufficiently high dose level.
For test chemicals of tow toEcily, a limit dose of 1000 mgfkg body weight per day (dietary
concentrations of approx]maZety 20,000 ppm [ 2% in the dietj for rats or 7000 ppm [ 0.7% in the
diecj for mice) is an adequate upper limit.
• The htstopatholo ’ data sbouk$ be summarized in at least two ways:
• A summary indMdual inh i data table: for example . list Individual animal number across the
top of the table versos tmdMdual lesions by tissue (can be further broken down by grade within
the tab I..., P—present. l mrnimM. 2—slight. 3smoderate, A—autolysls, X—noi remarkable,
l ir. k1& seedon , etc.) down the side of the table.
A sii y beam ill1 4 LI W table: for example. list dose group across the top of the table versus
indMdu tki ni by thius down the side of the table. Ii is 1 imponant to include number
of tissues m r i per dose level in this table. The table may aLso include separate tabulations
and total tabulations fur animals that either died or were sacrificed in moribund condition
during the study and fur inintil that were sacrificed on a predetermined time schedule.
• W ien evaluating body weight changes, include body weight gains in treated aninials when compared
to controls.
D-6
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
• Include food efficiency calculations (body weight gains versus food consumption).
• When appropriate and feasible, historical control data should be provided for neoplastic lesions
which were significantly increased in treated animals following administrauon of the test chemical
and for particularly unusual flOnneoplastic lesions which were identified in treated animals following
administration of the test chemical. The historical control data should be presented by individual
studies, tumor types, laboratories and sex (i.e. data presented as a range is not sufficient).
C. Combined c±zunic tyA) eni1 ty studies (&3-5)
The following supplemental guidance is provided:
• Data should be available to indicate that the animals were tested at a sufficiently high dose level.
For test chemicals of low toxicity, a limit dose of 1000 mg/lcg body weight per day (dieiaiy
concentrations of approximately 20,000 ppm [ 2% in the diet] for rats or 7000 ppm (0.7% in the
diet] for mice) is an adequate upper limit.
• The histopatholo i data should be summarized in at least two war:
- A summary individual animal data table: for example, list individual animal number across the
top of the table versus individual lesions by tissue (can be further broken down by grade within
the table, ic, P=present, 1=minimal, 2=slighi, 3=moderate, A=autolvsis, X=not remarkable,
1= incomplete section, etc.) down the side of the table.
- A summary lesion incidence table: for example, list dose group across the top 01 the table versus
individual lesions by tissue down the side of the table. It is important to include number
of tissue examined per dose level in this table. The table may also include separate tabulations
and total tabulations for animals that either died or were sacrificed in moribund condition
during the study and for animals that were sacrificed on a predetermined time schedule.
• When evaluating body weight changes, include body weight gains in treated animals when compared
to controls.
• Include food efficiency calculations (body weight gains versus food consumption).
• When appwp&re and feasible, historical control data should be provided for neoplastic lesions
which were ‘ Ignificantly increased in treated animals following administration of the test chemical
and for particularly unusual nonneoplastic lesions which were identified in treated animals following
administration of the test chemicaL The historical control data should be presented by individual
studies, tumor types, laboratories and sex (i.e. data presented as a range is not sufficient).
D-7
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December 24. 1989
PHASE 3 TECHNICAL GUIDANCE
W. TERATOLOGY STUDIES
A. Qarlflimdo
The discussion below clarifies Guideline Number 83-3 and tadlestes how EPA is applying the
guidance. The letter and number designations follow those in the guidelines.
U should be rea,gnized that this study is not a chronic or long term study and cannot be used as
such for risk assessment purposes although listed under tluJ heading in the Guidelines.
Teratolo , studies should more correctly be titled as Developmental Toxicity Studies as evaluation
of other manifestations of developmental toxicity are required to be assessed in addition to
malformations. This is also a necessary change in order to reflect that the subnutted studies are
consistent with the Agency’s risk assessment guidelines.
(A) When Reouired . Postnatal studies are presently being required by the Agency on a case-by-case
basis if it is deemed necessary to further evaluate any of the manifestations of developmental
toxicity. There are plans to make this a conditional requirement
(3) Puroose . This to ctiy study is designed to determine the potential of the test substance to
nduce any of the manifestations of developmental toxicity in the fetus which may arise form
exposure of the mother dunng pregnancy. The m nifesiauons include (1) death (2) structural
abnormality (3) altered growth and (4) functional deficiency. Functional deficiencies can
generally be investigated only in postnatal studies.
(C) Definitions . As noted earlier, the Agency is concerned with more than just teratogenic effects
and Ilteretore, the definition (or developmental toxicity needs to be provided.
(1) Developmental Toxicity, the study of adverse effects on the developing organism that may
result from exposure prior to conception, dunng prenatal development, or postnatally to the
time of sexual maturation. Adverse developmental effects may be detected at any point in the
life span of the organism. The major manifestations of developmental toxicity include: (I) death
of the developing orpasm . (2) structural abnormality, (3) altered growth, and (4) functional
def Iciency.
(D) Standard of the Test Methet In addition to the listed methodoIo ’, on a case -by-case basis the
Age y require $ postnatal evaluation. This may necessitate additional groups of animals
that aI J to deliver normally with o prtng evaluated to weaning or possibly to sexual
ma BebavloraWeve)opmenlal neurotoxicfly batteries may be included as part of this
ass ’ ee aewot dty discn sed below.
(E) Subsianc to be Testet Testing shali be performed in oral gavage studies with the technical
grade of the actIve In dient. In dermal exposure studies, testing is also preferred utilizing the
technical grade but a mpanying dermal absorption studies using both the technical grade and
formulations that are marketed may also be required in order to allow for meaningful risk
assessments to be performed. In addition, on a case-by-case basis, dermal developmental studies
using the formulations marketed may be required if dermal irritation is deemed minimal (after
consultation with the Agency). Only when no other alternative is available and only after the
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December 24, 1989
PHASE 3 TECHNIcAL GUIDANCE
Agency has been consulted, should dermal studies be conducted using an irritating formulated
product.
(F) Limit Test . If an oral gavage dose level of at least 1000 mg/kg produces no evidence of
developmental to city (even if some level of maternal toxicity is clearly present), studies at
ocher dose levels may not be considered. There is no limit test for other routes of exposure.
( G)Test Procedures
(G)(1)(iv) Number . At least 20 pregnant rats, mice or hamsters or 12 pregnant rabbits are now
required at each dose level and control group. Often 20 pregnant rabbits are necessary.
especially in a repeat rabbit study designed to claril the results of an initial study. The
objective is to ensure that sufficient numbers of litters are available to permit
meaningful evaluation of the potential developmental toxicity.
(G)(3)(ii) If a vehicle is used, its toxicological properties should be understood; it should not
induce developmental or reproductive toxicity. Dermal studies should not be performed
utilizing vehicles that are imcants unless all reasonable steps have been taken to reduce
or eliminate potential irritation to the maternal animals and the Agency has been
consulted prior to initiation.
(G)(3)(if I) To select the appropriate dose levels, a pilot or trial study is advisable using pregnant
animals.
(G)(3)(iv) Although the Agency requires the highest dose level to induce some level of overt
maternal toxicity such as weight loss, a is recognized that limiting maternal mortality to
less than 10% may not always be possible. Such studies have routinely heen accPpted
when mortality exceeds this level (but is less than 50%) and such severe effects (e.g. -
mortality) do not extend into other dose levels. Studies where mortatity may exceed
50% at the high dose level are considered on a case-by-case basis but dose related
mortality must never reduce the numbers of litters available at the middle or low dose
levels.
(G)(5) Administration of test substance . Oral gavage studies should be performed initially in
order to ascertain the potential of a pesticide to induce developmental toxicity. If any
studies are positive for developmental toxicity, and if worker exposure is anticipated,
dermal developmental toxicity studies and dermal absorption studies should be carried
out in order to allow the Agency to perform risk assessment. Before dermal studies are
thidated conairrence from the Agency should be obtained.
(G)(7)(i,fi) et, tJo of animals . Careful clinical examinations for signs of toxicity should be
m t on a daily and not a weekly basis.
(G)(7)(v) Measurements of food consumption should be carried out in all developmental toxicity
studies regardless of route of exposure.
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PHAsE 3 TECHNICAIt. GUrDANCE
(G)(7)(vf) Dams should be weighed at least every 2.3 days during gestation and at the time of
sacrifice, In postnatal studies, pups should also be weighed at Least at weekly intervals.
(O)(8)(i) Gross necmg y . At the time of sacrifice or death, dams should receive a thorough gross
necropsy. If animals are found dead or moribund, a veterinary pathologist should
specify the apparent cause of deathlmorlbundiry when possible.
(G)(8)(ii) Appropriate staining, glass plate techniques or other appropriate methodolo should be
employed to verify pregnancy and assist in accurate counts of early rcsorpt lons.
(G)(8) Better utilization and .nunatiOn of (vi) mouse, rat, and hamster and (vu) rabbit
fetuses involves dissection of the torso followed by cleanng, staining and then skeletal
examination of all fetuses. Half of the fetal heads are often dissected while the
remainder are cleared, stained and receive a skeletal exam. Other acceptable
methodolo , Involves a series of sections of the heads followed by clearing, staining and
skeletal mination. Care must be taken, however, (if these methods are to be
accepted) to assure that a thorough examination of soft tissues of the head has been
performed.
(H)(2) Evaluation of results . Historical data when necessary need to be provided for a period
of t years prior to as well as two years after the date of the study to be submitted
(when ‘ Ider studies are submitted) Data should be presented separately according o
the route of administration. In addition, individual study data (dated) as well as means
and tanges across studies need to be made available.
Studies must provide a NOAEL for developmental toxicity.
(H)(3) T t reoort . Although the litter is considered the most relevant unit for Statistical
assessment, data should also be presented and assessed on a fetal basis.
The test report must Include all Individual animal data as well as tabulated summary
tables.
B. Future Ph .iqj
It has be reco.a1s 4 that the clmsleal studies do not allow for assessment of all aspects of
deveIopmen kki.), . For mpIe, almost nothing is learned relative to the potential for functional
impairment the osnr n derived litter. Hence, the Agency is more often requiring additional
postnatal i. 1 iaus . Somrlme , these investigations help in further defining observed findingi from
the more - J studies or, in the case of developmental acuroloxicaty testing, they may aid in
further understanding esnual flcrsvus system related developmentaj effects. Such Guidelines have been
developed in the Agency and the mggers (or requirement of these studies are currently being evaluated
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PHASE 3 TECHNIcAL GUIDANCE
V. REPRODUCTIVE AND FERTILITY EFFECTS STUDEES
(i 5flfiC3tR)i
The discussion that folloM clarifies Guideline Number 83-4 and indicates how EPA is applying the
guidance. The letter and number designations follow those in the guidelines.
(E)(4)(II)(A&B) Test Procedures . (4)(ii)
After the conditions of (7)(i)B) are met, at the time of sacrifice, reproductive and target organs
from all parental males in the dose groups and controls should be weighed and preserved for
histopathologi I examination.
(E)(7)(iv)(A&B) Standardization of Litter Sizes .
Culling litters to 4 males and 4 females is not mandatory.
(E)(9)(i) Gross Necrovsv .
Weights of the reproductive organs and identified target organs and pituitary (absolute and
relative to body and to brain) should be provided.
A gross necr’psy is required for:
1 All parental animals which are either scheduled sacnfice or are found dead or sacrificed
moribund.
2. All pups found dead prior to day 21.
3. All weanlin not selected as parental animals of the next generation.
4 Not generally required for pups culled on day 4. However, should positive
developmental effects be noted form other studies, necropsies should be considered.
Generally, if developmental toxicity studies have been conducted, findin from these studies
need to be made available to the investigators of the reproduction study prior to the necropsies of
offspring.
(E)( 10) Clarification of HIEooathoIo y Reuuirements
i-iistopatho b required for
1. Reproductive and target organs of all control and high dose P1 and Fl parental animals
2. Organs showing patholo in the high dose level should then be examined in animals in
the mid and low dose groups.
3. Although the Guidelines are not clear on required histopatholo , for animals that died
or were saaiflced moribund, reproductive and target organs form these animals must be
saved (unless autotysis prohibits) and receive histopathological examination. This has
been standard policy for all toxicity studies.
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PHASE 3 TECHNICAL GUtDANc
4 F.nmIn th)fl of day 21 o pring not seIecte as parental animals should also receive
hiscopathologi c minauon fOllowing the pattern required for parental animals, this
is the only opportunity foP minanon of sexually immature of pnng in the
reproduction study.
5. Histopatho1ogica e minacfon of pups culled on day 4 Li flOt required.
B. Limit Test
The public has commented that there should be a ‘linist test’ for rcproduc ion studies (834). In
response. it is suggested that the same limit test for chronic feeding studies be applied to reproduction
studies, I.e. 1000 mgfKg/day.
C. Evaluation of the Male Repiodutilve S tem
Over the last few years OPP has more and more routinely required additional testing relative to
toxic effects of pesticides on the male repwductive system (other than what has been required by the
1982 Guidelines). Usually this has resulted from the generation of separate male studies (requiring the
use of additional animals) since such data have rarely been available form existent reproduction studies.
i a result, delays in some registrations and tolerances until these data are generated have been noted.
These data are essential (during the risk assessment process) in allowing the Agency CO assess the
potential for sex specific differences in to,acity to pesticides and allowing it to regulate appropriately. In
addition, when males from the reproduction study are utilized, no additional animals are utthzed in
order to obtain Ihcsc data. Therefore, the following procedures should be incorporated as part of any
newly generated re1 roductIon studies in order to avoid prior difficultics:
I Organ weights • (a) both testes should be weighted individually and identified as right or left.
(b) epididymis - whole and cauda, (c) a ssory sex glands (semInal vcsicl s with and without
their fluid contents and prostate , and (d) pituitary.
2. Sperm - left testis (a) spermatld count, (b) total cauda epididytnal sperm count (C) assessment of
morpholo i and motility, and (d) epididymal fluid cwnjned for debris and unexpected cell
t C3 Video tape recor are maintained as raw data allowing verification of reported results.
Investigator methadolo p used for sperm motility assessment must be clearly presented in the
submitted report.
3. Hislopatholo ,. the right testis Is d In 10% neutral buffered formalin for at least 48 hours
prior to embedding in glycol methacrytate, sectioning at 2 microns and staining with PAS
te’nan 4I. . The right epididymia is fixed and retained for possible future emmination.
D. Futiuu
OPP has 1 4ered the me of a modified version of the continuous breeding protocol to be
allowed as an alternative to the 2-generation study spe fied In the Guidelines. Ai per policy
established in a previous Agency worbbop, reproduction studies must have ruo generations. Hence
the Primary Continuous Breeding study modificatloas under consideration by the Agency deal with
the addition of a second generation to the Continuous Breeding Study. Other considerations
include the species to be tested (mouse or rat and the advantages and disadvantages of using each
as apparent from testing available to date.
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PHASE 3 TECHNiCAL GUIDANCE
At this point, the Office of Pesticide Programs is considenng holding a workshop in order to
further develop this protocoL
Vt NEUROTOXJCflY STUDIES
A Background
In a petition flied on February 12, 1987, the Center for Science in the Public Interest (CSPT) and a
group of states, public interest groups, and professional societies sought to have EPA. under FIFRA,
issue testing methods necessaty to fully assess the neurotoxic and neurobehavioral effects of pesticide
active and inert ingredients and require that registrants undertake such tests for pnority chemicals
detailed herein’.
They specifically proposed that EPA use the neurotoxicity guidelines developed by the Office of
Toxic Substances (U.S. EPA, 1985,1988).
OPP convened a meeting of an expert subpanel of the FIFRA Scientific Mvisory Panel on October
15-16, 1987 to review its proposed response to that petition (U.S. EPA. 1987). The Subpanel was
requested to address a set of issues concerning when such testing might be appropnate to require for
chemicals subject to FIFRA. In general, the SAP subpanel endorsed the use of:
• an observational battery, motor activity, and neuropatholo as an acute and subchronic screening
battery,
• further testing using tests of learned (operant) behavior, sensory testing, or other types of tests, or
chronic testing, based on the results of the screening tests, and/or based on chemical class;
• developmental neurozoxicity testing under some circumstances based on chemical class, hormonal
action, or data on adults.
The SAP Subpanel requested that the test guidelines for these data requirements be brought back to
the SAP for review. OPP is revising its data requirements and testing guidelines for neurotoxicuy testing.
The SAP reviewed the new test guidelines on September 28, 1989. Among the revised guidelines is a
new guideline fur testing of Organophosphorus Substances for Delayed Neurotoxicity.
EPA is d ) developing a Risk Assessment Guideline for Neurotoxicity for use throughout the
Agency. This 4iM ”.nt will discuss general policy for the assessment of all types of neurotoxicity data,
but will draw o the types of data in the test guidelines as illustrative of many types of neurotoxicity.
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PHASE 3 TECHNICAL GUIDANcr
B. Planned Ø L i4j i Rcv o
81.7 l te l la,md N o tT - ___
82.590 Osy Deia ed NeuroU ty - hen
OPP is planning to adopt a revised guideline for delayed neuro1o city. This guideline wifl require
range finding, acute, 4 day, and 20 day dosing regimens in hens in combination with assays of neurotoxic
esrerase (and aeetylcholincsterme) to sexeen for clii i effect. ft will provide more effective and conv,ncing
negative data sets than the previous requirement, facilitate interpretation of data, and improve the
determination of NOELi (or positive agents as well as an indication of whether furTher exposure would
suggest an even lower NOEL.
The new guideline will replaec GuidelInes 8 1-7 and 82-5, Acute and Subchronic Delayed
Neurotoxicicy of Organophosphorus Substances.
8 1 - i Aoite ncurot dty- rat
An acute neurotoxicity battery (Functional Observational Battety, Motor Activity, and
t4europatho1o l) is proposed for the pnmary route of exposure. e.g. oral (or food use, inhalation for
fumigant.s. etc.. The battery may be associated with any one of the 3 acute toxtcny stud es, or, preferably,
conducted as a separate szu4
82-5 Sub roeIc u’. ’zI .’ty -rat
The subchronn neurotoxicity battery (same tests as the acute battery, listed above) is being pr.posed
(or the route, of pnn..irv exposure for the use pattern and should generally be the same as the route for
the acute study.
l-l Chronic ‘ing - __
For new pesticides, if a chronic totcfty study In rats is needed, it should include a neurotoxiuty
battery (as described above). For old chemicaLs, adding a neurotoxicity battery to chronic studies will
probably not be req lrnd.
Develoomcncal N .Iu& I . g
A developmental neurot*udcfty study in rats may be required for a variety of chemicaLs registered for
different use pnflsI . It isus of exposure to dams through pregnancy and of pr1ng through
weaning. Test1 ,tlI 1a ude the tests in the screening battery, as well as developmental Landmarks, and
tests of Iearn$ me.oiy. This end point will be evaluated in the postnatal developmental toxicity
testing dtsu -
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PHASE 3 TECHNICAL GUIDANCE
va MIJTAGENICfl’Y STUDIES
The only major deviation from the 1982 version of the Assessment Guidelines 84-1 and 84-2 is
concerned with the adoption of specific protocol guidelines. The 1982 version suggests reference
documents (e.g. Gene-Tox reports) for guidance on protocols. Although the Gene-Tox effort is
supported by the USEPA, these documents are not official EPA policy. Protocol guidelines have been
written by OTS and penodically undergo official revision and review (found in 40 CFR Part 798). OPP
has informally adopted these protocol guidelines for the conduct of mutagenicity tests and this is a
deviation which is regularly discussed with registrants. The upcoming Subuivision F revised guidelines
which were reviewed by the SAP in September, 1989, will formally adopt these protocol guidelines as
OPP policy. The adoption of these protocol guidelines alleviates some of the concerns and questions
that have been brought up over the past few years. For e inple, i) it describes criteria for top dosing
in tests, II) statistical methods when appropriate is slated (many of the protocol guidelines state these
are necessary, but for some studies, such as the Salmonella assay, there are no consensus statistical
methods to use), iii) for the micronucleus test, the original ‘Schmid protocol’ (2 doses, 24 hours apart
with lull 6 hours after last dose) is no longer an acceptable protocol.
For the future, the mutagenicity section (Series 84) of the Subdivision F guideline will be revised.
The revisions have been written and were reviewed by the OPP’s SAP. There was also a public review
period in association with the SAP review. The SAP met at the end of September, 1989. EPA is
reviewing the SAP and public comments and the Agency will finalize the Subdivision F guideline for
mutagenicity in the near future.
vEil. METABOLISM SflJDIES
A metabolism study (85-1) may be performed and submitted in two parts:
1. Kinetici by individual dose
2. M tabolism by individual dose
IX. DERMAL ABSORFI’ION SflJDIES OF PESTICIDES (RESERVED)
There is presently no guideline. The Agency is planning to produce a draft guideline (85-3) and
supporting documents in 1990.
Registrants ate advised to ansuft with the Agency before performing a dermal absorption study
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PHASE 3 ThC1* CAL GUIDA$CE
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PHASE 3 TECHNICAL GUIDANCE
OVERVf w DOCIJME rr FOR SUBDIVISION 0
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Decembcr 24, 19
PHASE 3 TECHNICAL GUIDAt IC 4 .
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December 24, 1989
PHASE 3 TECHNICAl GUIDANCE
OVERVIEW OF SUBDIVISION 0:
RESIDUE CHEMISTRY GUIDELINES
This document provides an overview of the technical guidance for Subdivision 0. Table E- 1 lists
EPA publications pertaining to Subdivision 0 that are available through the National Technical
Information Service (NTIS). The first document on the list, the Pesticide Assessment Guidelines (PAG,
sometimes referred to simply as rThe Subdivision’), provides general guidance. It is suppIemente 1 by
the Data Reporting Guidelines (DRG) published as a series of addenda. Table E-2 sho where to
locate particular topica in the Pesticide Assessment Owdelines and Addenda. The body of the
document revie major changes since the guidance was originally published in 1982; clarifies some
apparent incorisistenctes between the Pesticide Assessment Guidelines, Data Reporting Guidelines, and
Standard Evaluation Procedures; provides some new guidance; and indicates future directions that the
Agency is considering.
There are three sections in this document. The first one is brief and contains general points that
do not relate to any particular study. The second section addresses each residue chemistry study with
(1) our responses to public comments on Inconsistencies and (2) changes or clarifications that l ave been
made in the guidelines since 1982. The third part of the document points out future changes being
considered for a thorough revision and updating of the Guidelines.
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December 24, 198
PHASE 3 TECHNICAL GUIDANCE
Table E 1. EM GUID.4NCE LYXUMENTS FOR SUBD(’v7S!OW 0
NTIS Number EPA Number
PESTICIDE ASSESSME T OUID JNES (PAO)
Residue Chemistry, SubdMsioa 0 PB83 -153981 540i 0 9-82-023
DATA REPORTING GUIDW.24!S (DRO)IADDENDA
Residue Chemistry.Mdendum 1 PBS6-203734 540 1 09.86-148
Residue Cbeirtistrj.Mdcndum 2 P386-248192 540,09-86-151
Residue Chemistry-Mdcndum 3 P 38 120864 1 540,09-87-199
Residue Chemistry-Addendum 4 P388-111270 540i09-88-0 04
Residue Chemistry-Addendum S P888.124003 540i09 -88-008
Residue Chemistry-Addendum 6 P388-191713 540109-88-049
Residue Cheinisuy-Mdendum 7 P389 .124598 540i0 9 - 89 O 9
Residue Chemistry-Addendum 8 PB89424606 540109-89-010
STANDARD EVALUATION PRO URES (SEP)
Analvucal Methods/Residue Chemistry PB90-I 3284 540-09-8 -062
Directions for Use P888-2 -3225 540iO9 ,1,6. 144
Magnitude of ihe Residue: Crop Field Trials P886-L942E 53’),1)9 8 {J21
Magnitude of the Rsidue: Processed Food/Feed Studies PB88-243209 .“4U 1)9- 6. 145
Qualitative Nature of Residue: Metabolism in Food Animals P890-1 (3292 541),09-8’JM6 I
Quaiitative Nature of Residue: Plant Metabolism P889-100333
Residues in Meat. Milk. Poultry and Egp: Dermal Trcairnents P388-233217 54ui09-8S- 2
Speci4(ty Applicatkn%s PB88-244454 54O109.8i - 142
Storage Stability Study/Residue Chemistry PB9O- 103276 540)09.*9M63
OTHER GUIDANCE
Position Document: Storage StabIlity P388-112362 540109-88-002
E -2
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PHASE TECH\tCAL CLiDA\’
Table E-2. WHERE TO FIND !NFQRM4 T!OV IN SUBDII’7S10 V 0 GUID -1 VCE DOCU tIE VTS
TOPIC LOCATION
Page in PAG Addendum Number
170 SCOPE OF DATA REQUIREMENTS 1
(a) General I
(b) Petitions for Tolerance
(C) Minor Changes in Use Pattern 1
(d) Food Use/Non-Food Use Determinations 1
(e) Tobacco Uses 2
(1) Aquatic Uses 2
171-1 LIST OF REQUIREMENTS 2
171-2 CHEMICAL IDENTITY 3
(a) Active Ingredient 3
(b) Inert Ingredient 4
171-3 DIRECTIONS FOR USE 4 6
(a) Application Directions 4
(1) Field and Orchard Crops 4
(2) Animal Treatments 5
(3) Fumigants 5
(4) Aquatic Uses 6
(5) Foreign Uses 6
(6) Food Handling Establishments 7
(7) Agricultural Premises 7
(8) Miscellaneous Applications 8
(b) Restrictions 8
171-4 RESULTS OF TESTS ON THE AMOUNT OF RESIDUE REMAINING 9 2, 3, 4. 5. 7, 8
(a) Nature of the Residue 9 3
(I) Characterization of the Total Terminal Residue 10 3
(2) Nature of the Residue in Plants 11 3
(3) Nature of the Residue in Livestock 12 7
(b) Analytical Methods 13 2
(1) General Requirements 13
(2) Validation of Method by Petitioner 14
(3) Extraction Efficiency 14
(4) Determination of the Total Toxic Residue 15
(5) Requirements for Regulatory Methods 16
(c) Magnitude of the Residue 17
(1) General Considerations 17
(i) Residues Determined 18
(ii) Sampling 18
(2) Design of Residue Experiments 19
( I) Field Studies 19 2
(ii) Fumigation Uses 20 5
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PHASE 3 TECH\ICAL GUIDA \CE
(iii) Stow Release Encapsulated Formulation Uses 21)
(iv) Processed Food/Feed Studies
(3) Meat, Milk, Poultry and Egg Feeding Studies 22 8
(i) Residues in Feed Items 22
(ii) Direct Animal Treatment 24
(iii) Agricultural Premise Use Studies 24
(4) Potable Water, Fish and Irrigated Crop Studies 25
(5) Food Handling Establishment Uses 27
171-5 PRACTICAL METHODS FOR REMOVING RESIDUES THAT
EXCEED THE TOLERANCE 30
171-6 PROPOSED TOLERANCES 30
171-7 REASONABLE GROUNDS IN SUPPORT OF THE PETITION 31
171-8 EXEMPTIONS FROM THE REQUIREMENT OF A TOLERANCE 31
(a) Active Ingredients 31
(b) Inert Ingredients 31
171-9 TOLERANCES FOR FOREIGN USE 32
171-10 ROTATIONAL CROP TOLERANCES 32
171-11 TOBACCO USES 33
171-12 FOOD USE 1 NON-FOOD USE DETERMINATION
DATA REQUIREMENTS 5
(a) Seed Treatments 34 5
(b) Crops Gro n for Seed Only 35 5
(C) Fallow Land 35
(d) Non-Bearing Crop Uses 35
171-13 SUBMIUAL OF ANALYTICAL REFERENCE STANDARDS 36
171-14 SPECIAL CONSIDERATIONS FOR TEMPORARY
TOLERANCE PETITIONS 36
171-15 PRESENTATION OF RESIDUE DATA 37
171-16 TRANSLATION OF DATA 37
Table I. Categones and Representative Types of
Food Establishments
Table II. RAC’s, Processed Commodities, Feeds and
Percents of Livestock Diets for all Crops
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PHASE 3 TECHNICAL GUIDANCE
GENERAL POINTS
Typing errors and inadvertent omissions in the text portion of the 1982 Guidelines-Subdivision 0 are
shown on Attachment 1 (Errata Sheet) Revisions that have been made to TABLE 11 of Subdivision 0
are shown on Attachment 2. The latter contains both inadvertent errors in the original published
document as well as revisions such as the addition of new feed items, changes in the restrictability of
livestock feed items, and lists of milled products for grains. [ Note that guideline clarifications and
changes in policy in olving the text portion of Subdivision 0 are discussed below under “Individual
Studies ‘1
Public comments on each type of residue cherriistiy study included ‘not enough guidance” EPA
RESPONSE Some areas will be discussed in more detail in a future thorough makeover of Subdivision
0 At this time we emphasize that any future studies should be conducted in such a manner that all
the acceptance criteria are met. These criteria are included elsewhere in the FIFRA ‘88 technical
guidance package and represent the key features of an adequate study
Another commenter noted that some DRG’s have experimental results documented in the “Methods’
section with the ‘Results’ being a summary of these data ‘Does EPA accept that a more logical
approach is to keep all experimental data within the ‘Results’ section, and to leave the summary to the
‘Conclusions’ section’” EPA RESPONSE’ Yes, we concur this is a more logical approach and would
certainly have no objections to such a format. However, as long as reports are clearly written and
contain the information noted in the DRO’s, either of these formats is acceptable.
Registrants alse’ tnmmented on the location of figures and tables ‘Does EPA accept thai figures
and tables are more easily accessible to readers when presented at relevant positions in the bulk of the
report, rather than grouped together in a separate section at the end .. “ EPA RESPONSE’ As tong as
tables and figures hase accurate titles and are listed in the Table oE Contents. either of these tormats is
acceptable.
INDIVIDUAL STUDIES
§171-4 Nature of the Residue-Plants
Comments
Commenters have provided a series of comments on this study to which we will respond below. We
note that many of these comments are also applicable to the Nature of the Residue-Livestock study.
The next four paragraphs deal with comments on how the nature of the residue study should be
conducted
COMMENT: The requirement to identify residues down to the 0.01 ppm level is ‘technically and
scientifically unsound, difficult (or impossible) to achieve,’ and introduces new requirements. EPA
RESPONSE: We agree that identification down to the 0.01 ppm level is difficult, but efforts to
characterize such residues should be made. As noted in a recent Agency memo (ATTACHMENT 3),
residue levels or trigger values have been proposed to the Agency by Ciba Geigy Corporation for the
degree of characterization necessary for radiolabeled residues. These trigger values (0.01, 0.01-0.05,
>005 ppm) are rough guides that registrants can use in their metabolism studies. However, as noted in
our 7125/89 memo, the degree of characterizanon should also depend upon how the application rate or
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December 24 I9 9
PHASE 3 TECHNICAL GUIDANCE
dosage in the metabolism study compares to actual use of the pesticide When low acti itv is found in
crop parts or animal tissues following highly exaggerated application rates, characterization requirements
should be less stringent
COMMENT. With respect to the determination of total radioactivity in a plant part, it is difficult to
obtain a representative subsample that will give accurate total 14-C by combustion for samples where
the residue is not evenly distributed or which have a high water content. “For these types of samples.
would it be acceptable instead to use a combination of extraction and combustion in order to determine
the total residue? Since the weighed subsample is extracted by maceration. and the supernatant is
separated by centnfugation, there are no losses due to ‘work-up’ Radioactivity in the liquid extract is
determined by USC, and radioactivity in the solid residue (which will be much more evenly distributed
than in the original sample) is determined by combustion and LSC.”
EPA RESPONSE: Yes, we consider this procedure acceptable for the types of samples noted.
COMMENT One registrant questioned why both polar and non-polar solvent systems should be
used in thin layer chromatography of radioactive residues. “Should not the polarity of the solvent
system be governed by the polarity of the compounds being analyzed?” EPA RESPONSE. Yes, the
solvent polarity should be adjusted to the compounds of interest. We note that our copy of the SEP
does not state “polar and non-polar solvent systems”, but instead “at least two different solvent systems”’.
Our main concern is that the residues be identified by matching Rf values in at least two different
solvent systems if TLC is the only identification procedure being used. If possible, we really prefer that
two separate techniques (e.g., TLC and MS) be used to identify residues (as opposed to TLC with two
solvent systems).
COMMENT An’)ther comment questioned the need for frequent “interval sampling of protein.
starch, ligrnn. and cellulose fractions.... Until a study is completed it is impossible to know whether
bound residues of 1O% are likely to occur.” EPA RESPONSE. Only in those crop parts wherein
toc or O 1 ppm ot residue is bound does the registrant need to employ hydrolyses (acid, base.
enzyme) in an attempt to release such activity. If this activity can not be identified as bound forms of
the parent pesticide or closely related metabolites, attempts should then be made to show that the
acti tty is associated with natural components such as starch or lignin.
The next five paragraphs address comments on the reporting of plant metabolism studies.
COMMENT: “The specific activity should be reported as LCi/mg instead of dpm/g or Curies/mole.’
EPA RESPONSE: Any units that would permit our calculation of ppm (mg/kg) radioactivity using
reported counts (dpm, dpmlg, etc.) are acceptable.
COMMENT: “Reporting data as total radioactive counts is cumbersome, redundant, and
unnecessary.a EPA RESPONSE: Sufficient information on counts should be provided so that we can
verify the ppm reported for crop parts, animal tissues, and the various chromatographic fractions
thereof. If more convenient, another unit such as dpm/g may be used. Regardless of the unit used, a
sample calculation should be submitted showing how the analyst arrived at “ppm” from the experimental
data.
COMMENT: “Photographs of TLC plates should not be required. Chromatograms provide much
more usable data.” EPA RESPONSE: We agree ‘chromatograms’ provide useful data and consider a
TLC plate an example of one. Photos of TLC plates or their autoradiograms that were critical to the
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identification of residues should be provided If HPLC coupled to a detector capable of measuring
radioactivity was emplo ed, then appropriate liquid chromatograms should he submitted Regardless of
the chromatographic technique used. chromatograms showing the behavior of the analytical standards
should also be included in the report
COMMENT. “The EPA asks that the total radioactivity in each plant part should be expressed as
ppm equivalents of the total recovered plant radioactivity at the time of sampling or harvest For many
crops, e g apples. it would be extremely difficult to determine the ‘total plant radioactivity’ Does EPA
require us to do this, or is it not more acceptable to express the residue in terms of Bq/ug, or ppm in
crop parts of specific interest onIy EPA RESPONSES At a minimum registrants should report the
total ppm radioactivity (usually in ppm equivalents of parent pesticide) for each crop part that could be
used for food or feed For those crops where the activity is measured in all plant parts, it would be
useful to report the % of total plant activity in each part, but this is not required
COMMENT: Enforcement methodology validation should not be reported in the nature of the
residue studies, but rather in the magnitude of residue studies EPA RESPONSE: We ha e no real
preference on where the radiovalidation of analytical methods should be reported. It could be part of
the report on the analytical method, part of the plant/animal metabolism study, or stand by itself as a
report The cover letter or summary of the full data package should indicate where it has been placed
§171-4 Nature of the Residue-Livestock
Comments
Many of the comments discussed above on the Nature of the Residue-Plants study are also
applicable to the live wck metabolism study
Clarifications and Re”isions
Registrants should note that the guidance on material to be fed in livestock metabolism studies is
incorrect The guidelines as published state that ‘The material fed should simulate the terminal residues
in feed items as closely as possible....’ This implies the studies should be carried Out by dosing the
animals with a mixture of radiolabeled chemicals. If animals are dosed with a mixture of parent and
plant metabolites, one will be unable to determine whether plant metabolites are also animal
metabolites. Therefore, the guidelines need to be revised to state that the animal metabolism studies
should reflect feeding of one compound, usually the parent If the plant metabolites are also found to
be animal metabolites, then additional metabolism studies involving dosing with the plant metabolites
will not generally be required. However, if a plant metabolite comprises a major portion of the residue
on feed or is not found to be an animal inetabolite, additional animal metabolism studies involving
dosing with the plant metabolite may be required.
Additional clanfication on when and how to conduct livestock metabolism studies was issued
recently (ATTACHMENT 3). As noted in the SUMMARY at the end of that memorandum, livestock
metabolism studies are now required whenever a pesticide is to be used on a crop having a livestock
feed item in Table II of the Residue Chemistry Guidelines. (Registrants should refer to the original
(1982) Table U as well as ATTACHMENT 2 to this Overview to determine if the crop of interest has a
feed item.) This memorandum also provides trigger values (ppm radioactivity) for the degree of
characterization needed in metabolism studies. These are applicable to both plant and animal
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metabolism studies and are also discussed earlier in this Overview ” under a comment on the Nature of
the Residue-Plants study Refer to AT1’ACHMENT 3 for more details
§171-4 Analytical Methods
Comments
COMMENTS The statement that the enforcement method should “not require the use of an internal
or procedural standard” is unacceptable EPA RESPONSES At this time we do accept the addition of
an internal standard to the final extract just prior to injection to serve as a calibration for retention
times However, the use of an internal standard throughout the entire procedure to correct for
recoveries is not acceptable unless data are available on numerous samples of each matrix to show
analyte and internal standard behave identically in each step [ We are presently working with FDA and
USDA to define what is a practical and acceptable enforcement method. This definition will become
part of a future revision of Subdivision 0 It is possible that the policy on internal standards will
change as a result of this project.J
COMMENT One commenter noted that a majority of their parent methods meet the requirement
of a maximum of 24 hours for completion. However, many metabolite methods will not. “It is not in•
the interest of any company to develop methods which are more time consuming than necessary. This
appears to be an ‘inappropriate trigger’ to be applied, and methods should be assessed on a case-by.
case basis” EPA RESPONSE: The guidelines presently do include a case-by-case evaluation. “Methods
taking longer than 24 hours will be considered acceptable on a case-by-case basis for minor metabolites
or residues that are ri n acutely toxic.” [ This issue is also being examined in our work with FDA and
USDA on defining a practical enforcement method.J
COMMENT The SEP implies that ‘aU matrices for which tolerances are proposed should be tested
in the analytical method validation and recovery data etc. should be provided....Would it be possibIe to
clarify that data for the appropnate typical crop types, e.g root, leaf, oil and grain, would be sufti ient? ”
EPA RESPONSE: The analytical method should be validated on each crop for which residue data are
generated and a tolerance is proposed. In the case of crop group tolerances, the method needs to be
validated on only the representative crops for the group. With regard to where the validation data
should be reported, we wish to point out that the report submitted on the method itself needs to
include recovery data on only representative crops such as noted in the comment. However, in the crop
field trial reports additional validation data should then be provided on any crop that was not tested for
the analytical method report.
Clarifications and Revisions
The availability of an addendum to the Subdivision 0 Guidelines on multiresidue protocols was
announced in the Federal Register in 1986 (Vol. 51, No. 18’7, 9,26/86, page 34249). Registrants are now
required to test pesticides for their behavior through the multiresidue procedures used by FDA for
monitonng residues [ 40 CFR 158.125(b)(15)J. The initial protocols were designated by Roman numerals
(I-IV) and made available through the National Technical Information Service (NTIS) as Order No. 86
203734/AS. However, FDA recently published a revised version of these protocols as Transmittal 89-1
for the Pesticide Analytical Manual Volume I (PAM I). As part of this revision, Protocol II (the
Storherr method) was removed as FDA no longer routinely uses this method. Also, a protocol was
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added to cover acids and phenols through PAM 1 221 1 An updated version of the acid’phenol method
in the Transmittal uses ion pair alkvlation in place of diazomethane In addition, to a oid contusing
protocols and reporting forms with earlier versions, the protocols are now designated by letters (A. E)
instead of Roman numerals Any future testing of pesticides for multiresidue behavior should use these
new protocols.
[ Ordering information for Pesticide Analytical Manual NTIS phone number 703-487-4600 Order
Numbers are PB 889 [ 17 99 for Volumes I + II, PB 889 118 99 for only Volume I, and PB 889
119 99 for only Volume II
Another requirement instituted since the 1982 publication of Subdivision 0 is the independent
laboratory confirmation of tolerance enforcement methods. Pesticide manufacturers were informed of
this requirement, which went into effect on 8/1(89. by PR Notice 88-5 issued 7/15/88 New tolerance
enforcement methods must have been successfully tested by an independent laboratory prior to
submission to the Agency This requirement was initiated to ensure development of clearly written,
complete descriptions of analytical methods
Registrants should be aware that a strong justilication is needed for the use of either diazomethane
or benzene in an analytical method A safer methylating agent or solvent, respectively, should be
sought If one can not be found, documentation should be provided supporting the need for use of
these hazardous reagents
§ 171-4 Storage Stability
Only limited discussion of this topic is present in the original Residue Chemistry Guidelines (page
19) However, it will become a separate line item in an upcoming revision of 40 CFR 158. In addition,
the Agency published a Position Document on the “Effects of Storage (Storage Stability) on Validity of
Pesticide Residue Data’ in August 1987. This document, available through NTIS Order No PB 88-
112362. gi es more details on the design and reporting of storage stability studies
Comments
COMMENT. “Storage stability studies conducted on any member of a major crop group (root, leafy
‘vegetable, cereal, oil) should be sufficient to satisfy the data requirement for each crop belonging to the
group” EPA RESPONSE: As stated in the aforementioned Position Document, “Translating a storage
stability study from one commodity to another will be considered appropriate only if both commodities
are related (e.g., in the same crop group), and if the experimental design is considered appropriate to
current considerations” The term “crop group” refers to the groups listed in 40 CFR 18034(f).
However, some combining of these groups into larger ones would generally be acceptable. Examples
might be combining (1) leaves of root and tuber vegetables (group ii) with leafy vegetables (except
Brassica) (group iv); (2) pome fruits (group xi) with stone fruits (xii); or (3) forage, fodder, and straw of
cereal grains (group xvi) with grass forage, fodder and hay (group xvii).
COMMENT. “If storage stability has been shown in the raw agricultural commodity, storage stability
studies in each of the processed fractions should not be required.” EPA RESPONSE: Since some
processed fractions (e.g., oils, fruit juices, soapstocks) have matrices quite different from the starting
r a c, storage stability data are required for byproducts. However, once storage stability has been
demonstrated in a few representative crops and their byproducts, additional, data would not be needed,
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provided storage conditions and duration are similar Suggested representative crops ould be an
oilseed (e g. soybeans), a fruit (e g, citrus). and a non-oily grain (e.g wheat)
Clarifications and Revisions
For more details on storage stability requirements, registrants should refer to the aforementioned
Position Document as well as the Data Reporting Guideline and Standard Evaluation Procedure
§ 171-4 Crop Field Trials
Comments
COMMENT. Specific tables on geographic locations and recommended minimum samplings
(size/weight/technique) should be included in Subdivision 0. Such tables are included iii the Guidelines
for Conducting Agricultural Chemical Residue Field Trials in the USA EPA RESPONSE: Such tables
will probably be included in a future version of Subdivision 0 With regard to locations for field trials,
for the time being registrants may refer to ATTACHMENT 4 to this document We remind petitioners
that tolerances may be based on geographically limited residue data for minor uses as described in the.
4,2/86 Federal Register notice (Volume 51, page 11341). With regard to sampling, registrants may
refer to the Food and Agriculture Organization (FAO) Plant Protection Bulletin, Vol. 29, 1981, page 12
for their guidelines on pesticide residue trials Our guidelines (page 19) state that the procedures
described in this publication should be followed in the design and execution of field trials.
Clarifications and Revisions
Registrants should refer to ATTACHMENT 2 to this document for changes in the r a.c ‘s associated
with certain crops For example, feeding restrictions are no longer considered practical for sugar beet
tops Therefore, residue data on the latter must be included in requests for pesticide uses on sugar
beets
There are four areas which will receive more attention in reviews of future studies plot size,
random sampling, sample size, and sample reduction. Petitioners should be sure that in future field
trials these four parameters are addressed in accordance with the aforementioned FAO Guidelines
(pages 14, 17, 19, and 21, respectively) and described in sufficient detail in the field trial reports.
We reiterate that a policy addressing tolerances based on geographically limited residue data for
minor uses was published in the Federal Register on 412186 (Volume 51, page 11341).
* 171-4 Proc sed Food/Feed
Comments
COMMENT. This section of the guidelines should include a table of concentration factors. EPA
RESPONSE: In the future we plan to include theoretical maximum concentration factors in an updated
version of Subdivision 0.
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COMMENT “We are concerned about the requirement for storage stability data on indi idual
processed fractions For field corn, which may be processed into 25 or so different commodities, this
represents an enormous and unnecessary waste of resources It would be far better to get agreement
that jf the storage stability of the residues has been proven in the RAC, then this should cover all
processing work.” EPA RESPONSE. As noted earlier in this document in the section on Storage
Stability, storage stability should be demonstrated in a few representative crops and their byproducts
(e g, soybeans, citrus, wheat). Once such stability has been shown, additional data would not be needed
on processed fractions, provided storage conditions and duration are similar With regard to field corn,
we do not require data on 25 fractions The processed commodities that need to be analyzed for grains
are listed below
Clarifications and Revisions
The original Table II stated that the processed commodities for grains included “milled products”,
but did not identify the latter by name Since that time we have developed lists of these products and
hate added grain dust as a byproduct of corn, rice, soybeans, and small grains. These changes are
included in ATTACHMENT 2 to this document In addition, all the processed commodities are listed
below for those crops which had the term “milled products” in Table II
Crop Processed Commodities
barley hulls, bran, flour, pearl barley, grain dust
buckwheat hulls, middlings, flour, grain dust
field corn wet milling.starch, crude + refined oils
dry milling-grits, meal, flour, crude + refined oils
grain dust
guar meal, gum
millet hulls, flour, meal, grain dust
oats hulls, flour, rolled oats, grain dust
rice hulls, bran, polished rice, grain dust
rye flour, rye feed (mix of bran, red dog flour,
and iniddlings), grain dust
grain sorghum flour, starch, grain dust
wheat bran, flour, middlings, shorts, grain dust
If an adequate processing study has been conducted on wheat, it would satisfy the requirement for
studies on barley, buckwheat, millet, oats, and rye. Guar, field corn, rice, and grain sorghum would each
require their own processing study.
The grain dust data are needed only in those cases in which detectable, primarily surface residues
are found on the grain. Early season herbicide uses usually result in low residues that would not be
concentrated on the grain surface. Therefore, grain dust data would seldom be required in these cases
At the other extreme, postharvest treatments of stored grains virtually always trigger the need for grain
dust data. Late season foliar uses to exposed grains such as wheat would also usually require such data
Clarification regarding the need for processing studies when no detectable residues are present on
the r a c. has been issued to EPA staff. The relevant portions of this guidance follow.
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‘If detectable residues are found on a crop for which Table I! of the Guidelines lists a processed
commodity, then a processing study is required. and if the data show a concentration of residues. then a
Food Additive Tolerance (FAT) is required. The processing study should use RAC samples bearing
detectable residues. However, if all RAC samples show no detectable residues as a result of the
proposed use, then the reviewer may be able to make a conclusion as to the need for processing studies
and FATs based on results of field trials carried out at exaggerated rates
If exaggerated rate data are available and these result in detectable residues, then these samples
should be used for a processing study If residues concentrate on processing, then the concentration
factor should be applied to the RAC tolerance level to arrive at the FAT level.
If exaggerated rate data are available and these result in no detectable residues, then no FAT is
required provided that. 1) the rate was exaggerated by at least the theoretical concentration factor, 2)
the data are sufficiently representative of important growing regions so that any reasonable potential for
detectable residues has been realized, and 3) the exaggerated rate was not unrealistically high. This
latter requirement arises over concern that if highly exaggerated rates are used the residue would not be
distributed in the same proportion as if a 1X rate were used. For example, a lox application of a
granular formulation may result in a larger proportion of the pesticide falling to the ground. The level
of exaggeration acceptable will depend on the use. For foliar application, a 5X exaggeration is likely to
be an upper limit, but for other types of application, e.g., pre-emergent, a greater exaggeration may be
acceptable.
If application of the highest practical exaggerated rate results in no detectable residues and the level
of exaggeration is Ics; than the theoretical concentration factor, then samples from this exaggerated rate
can be the basis of a processing study If no detectable residues are found in the processed product,
then no FAT is required. If the processed commodity contains detectable residues, then a FAT is
needed.
In cases here the RAC contains no detectable residues, the processing study will indicate only that
the minimum concentration factor is the ratio of the concentration in the processed commodity to the
limit of detection (LOD) in the RAC. The reviewer should evaluate all available data in determining
the appropriate concentration factor. This will include, at a minimum, the metabolism studies and the
chromatograms (or other raw analytical data) for the RAC samples. In some cases ii may be possible to
estimate residue levels from chromatograms where the response is below the limit of reliable
quantitation but indicative of a “true residue.
When using exaggerated rate data for consideration of FATs, reviewers should be careful not to set
a FAT that is too lugh based on data not reflective of real-world use. Reviewers should also be careful
not to require a FAT if there is no reasonable possibility that the processed product will bear residues
higher than the RAC. Occasionally, use of exaggerated rate data will result in no clear indication of
whether a FAT is needed, or, if it is needed, what level it should be. In these situations, considerable
judgment, based on all pertinent data, will be required.
Registrants should use the above guidance for determining when processing studies are required and
at what levels food or feed additive tolerances should be set.
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* 171-4 Meat 1 Milk. Poultry and Egg Feeding Studies
Comments
COMMENT There is no clear statement in the PAG that samples must not be composited (except
for milk) The SEP states that samples must not be composited except in the case where small sample
sizes are expected, e g eggs Three unique samples must be analyzed at dose level This is
more than we are currently doing for poultry transfer studies
EPA RESPONSE Three unique samples of milk. eggs, and edible tissues should be analyzed at each
dose level to show the variability of residues among different animals. In the case of cattle, this usually
means one sample per animal as three cows are generally dosed at each level. For poultry. eggs or
tissue samples from 3-4 birds may be composited to generate the three “unique samples for each dosage
group
COMMENT’ With regard to the SEP on dermal treatment of livestock, To expect analysis of a
formulation for impurities in the active ingredient is not responsible. Trace amounts of impurities will
not be significant in these studies. ... The concentration of the impurities in the active ingredient in the
formulation may therefore be calculated using the data in the confidential statement of formula for both
the active ingredient and the formulation.’ EPA RESPONSE. The request for impurity information was
intended to apply only to impurities which are actually regulated in the tolerance. Only a very small
percentage of pesticides have such impurities. In those instances in which the impunty is regulated, the
level of the impurity in the formulation applied to the animal should be measured to determine the
actual exposure to that component. If the batch of technical grade active ingredient used to prepare the
applied formulation wjs analyzed for the impurity of interest, the level of the impurity in the
formulation could be calculated using the degree of dilution of the technical.
COMMENT “The PAG states, ‘Analysis must be conducted on whole milk and eggs (yolk and
white)’ Does this mean analysis on yolk and white combined into one sample. or instead, separate
analysis on each pattY” EPA RESPONSE: The analysis should be conducted on the yolk and white
combined into one sample. They may also be analyzed separately provided the weights of each are
known so that the residue can be calculated on a whole egg basis.
Clarifications and Revisions
With regard to the possibility of waiving conventional livestock feeding studies when low residues
are present in feed items, registrants should refer to ATTACHMENT 3. In some cases the livestock
metabolism study may also serve as a feeding study and indicate that meat and milk tolerances are not
necessary
Registrants should refer to ATTACHMENT 2 for changes in the status of feed items. New feed
items such as grain dust and potato waste have been added to the table Sugar beet tops and wheat
straw are examples of crop parts which can no longer be restricted as livestock feeds by label statements
In the case of corn, some %‘s of the diet were discovered to be incorrect.
Registrants are reminded that the percents of the livestock diet in Table II of Subdivision 0 are on
a dry weight basis. Therefore, for feeds with high moisture content such as fresh grass the residue level
must be convened to a dry weight basis before utilizing the percent of the diet figures.
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The text in the guidelines addressing the duration of feeding studies has been revised The original
guidelines state that ‘Generally, animals should be kept on test until residues plateau in milk or eggs
If rio residues are detected in milk or eggs. animals should be kept on test for four weeks’ These
sentences should be replaced by ‘Animals should be kept on feed for four weeks. However, if residues
have not plateaued in milk or eggs by the end of four weeks, the feeding period should continue until a
plateau is reached.’ This change was made to (1) address the possibility that the plateauing of residues
in milk does not guarantee residues have plateaued in tissues and (2) to avoid situations wherein
registrants terminate a study well before 28 days and the Agency does not agree a plateau was reached
for milk or egg residues.
With respect to dermal treatment of livestock we generally do not consider preslaughter intervals
(PSI) longer than 3 days as being practical. In dermal treatment studies the animals should be sacrificed
within the PS! on the product label. However, as we have observed that residues may not peak in
tissues until a week or so after application, additional data reflecting longer PSI’s should be obtained to
establish the maximum residue levels for tolerances.
§ 171-5 Reduction of Residues
In recent years there has been much interest on using residues in food as consumed (‘anticipated
residues’) when conducting dietary risk assessments. Although there will be no further discussion of this
topic in this ‘Overview’, a chapter on ‘anticipated residues’ is being prepared for inclusion in the
FIFRA ‘88 Phase 3 Guidance Package. Registrants should refer to this chapter for additional
information on market basket surveys and studies on degradation in storage, cooking, peeling, and
ashing More detailed guidelines in this area are planned for issuance in mid-1990.
§171-12 Food Use/Non-Food Determination Data Requirements
One area in which numerous questions have arisen over the years is whether uses of pesticides on
crops grown for seed are non-food uses, which do not require tolerances. Our concern has been that
the crop may be di eited to food use before seeds are produced or that the seeds themselves may be
used for food or feed. regardless of label statements prohibiting such practices
The following two paragraphs outline the policy as published in the 1982 Guidelines. Please note
that these paragraphs replaced the first paragraph published on page 35 in the original document (see
AUACHMENT 1-ERRATA SHEET).
‘Uses on crops grown for seed only are considered to be non-food uses if (1) subsequent to
treatment the crop is not fit for consumption by humans and is not fed to livestock, and (2) there is no
likelihood of residues in crops grown from the harvested seed. Otherwise, such uses are considered to
be food uses requinng tolerances.
Examples of uses where the crop is considered unfit for consumption subsequent to treatment are
post-bloom applications to leafy vegetables and Brassica (cole) leafy vegetables, and desiccant uses on
okra and root and tuber vegetables. These uses would qualify as non-food uses provided there is no
likelihood of residues in crops grown from the harvested seeds. Factors affecting this include the level
of residues on the harvested seed, the half-life of seed residues, the weight of the seed in relation to
that of the subsequent crop, and the amount of residue uptake into the aepal portion of the crop.’
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In recent years we have accepted some uses which do not meet requirement (1) noted abo e If a
siate is willing to provide assurance that seed crops will not be diverted to food or feed and information
on acreage and cultural practices is provided, we will consider pesticidal treatment o [ crops grown for
seed as a non-food use under registrations The information on cultural practices should
emphasize those practices which distinguish the seed crop from the corresponding food crop and which
would render the seed crops commercially unsiable for food Examples of such practices might be
smaller crop spacing preventing adequate root formation and planting in a different season to encourage
bolting versus head formation
Once the State’s assurance and cultural practice information outlining a rationale as to why these
are non-food uses have been submitted, we will consider these uses on a case-by-case basis for 24(c)
registrations. Most cole crops, leafy vegetables, and root crops have a good chance of obtaining
pesticidal treatment of their seed crops on a non-food basis. Crops where the seeds themselves are
major raw agricultural commodities such as grains, beans, and peas have very little chance of such
registrations on a non-food basis. Alfalfa, clover and grass grown for seed are generally not considered
non-food uses since they can be cut for hay prior to going to seed and the seeds or seed screenings may
end up in livestock feed. However, we have recently accepted use on alfalfa grown for seed in
Washington State based on rules issued by the State governing the use of the seeds, seed screenings aiid
other crop parts
NOTE. FOR THE FOLLOWING RESIDUE CHEMISTRY GUIDELINE NUMBERS. NO SPECIFIC
COMMENTS WERE RECEIVED AND WE HAVE NO REVISIONS OR CLARIFICATIONS
§ 171-4 Magnitude of the Residue-Potable Water
§ 171-4 Magnitude of the Residue-Fish
§ 171-4 Magnitude of the Residue-Irrigated Crops
§ 171-4 Magnitude of the Residue-Food Handling
§ 171-6 Proposed Tolerances
§ 171-7 Reasonable Grounds in Support of Petition
§ 17 1-8 Exemptions from the Requirement of a Tolerance
§ 171-9 Tolerances for Foreign Uses
§ 171-10 Rotational Crop Tolerances
§ 171-11 Tobacco (See ATTACHMENT 1-ERRATA SHEET page 34)
§ 17 1-13 Submittal of Analytical Reference Standards
§ 171-14 Special Considerations for Temporary Tolerance Petitions
§ 17 1-15 Presentation of Residue Data
§ 171-16 Translation of Data
FUTURE REVISIONS OF RESIDUE CHEMISTRY GUIDELINES
This section of the Overvtew points out changes that are being considered for an updating of
Subdivision 0 in the next few years.
With regard to ruminant metabolism studies. significant differences have been observed between
goats and cows in the amounts of metabolites from some pesticides. Since cattle contribute much more
to the human diet, future ruminant metabolism studies may be limited to cows only. In addition, swine
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metabolism studies may be required in the future whenever a feed item for swine is treated with a
pesticide Presently, these studies are required only when metabolism in ruminants ditfcrs from that in
non-ruminan9 Future livestock metabolism studies may also have to include identification of residues
in feces and urine.
We are working with FDA and USDA to define what is a practical and acceptable enforcement
method As noted previously in this “Overview”, issues to be addressed include the length of time
needed for analyses and the use of internal standards.
Several projects are being conducted to update Table II Food processors have been surveyed to
determine the quantities of byproducts and their uses in animal feed Experts in animal nutrition will
be consulted to determine the percentages of assorted feeds in livestock diets. The gathered information
v ill be used to update the processed commodities and feed items in Table I! along with the %‘s in the
diet The practicality of feeding resinctions for crops such as small grain forages, the vines and hays of
peas/beans/peanuts. and sorghum forage and fodder will be investigated. We also plan to change some
of the names for raw agricultural commodities to eliminate confusion over what is to be analyzed For
example. the terms forage and fodder have different meanings depending on profession and/or location.
Words such as “green vegetative growth stage” may be used in the future. Theoretical concentration
factors for some processed commodities will also be in the new table.
Radiolabeled studies may be required to determine what degradates are formed by pesticide residues
upon cooking, canning, frying and other high temperature processes The latter occasionally result in
significantly higher levels of toxic residues with the best known examples being daminozide to UDMH
and the EBDC fungicides to E11J.
Future livestock feeding studies may need to include an additional dosage level below the so-called
1 level, which represents tolerance residues on all feed items. The additional dose will be used to
more accurately predict “anticipated” or average residues in animal products. Although meat/milk
tolerances would still be based on the lx level, dietary risk assessments would usually use the results of
a lower feeding level.
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ATTACHMENT 1-ERRATA SHEET
ATTACHMENT 2-’REVISIONS IN TABLE II
ATTACHMENT 3• Guidance on When and How to Conduct Livestock Metabolism
Stud ies”
ATTACHMENT 4- SUOGESTED STATES FOR CROP FIELD TRIALS”
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ATTACHMENT I
Pesticide Assessment Guidelines
Subdivision 0
Residue Chemistiy
ERRATA SHEET
This Attachment lists the typing errors that were made in the text portion of the original guidelines
in 1982 Note that typing errors were also made in Table II of the original Subdivision 0 These are
included in Attachment 2 to this “Overview” of the Residue Chemistry Guidelines along with other
changes made in Table II since 1982 (e g.. sugar beet tops may not be resincted as a feed item, potato
waste is now considered to be a feed item). Guideline clarifications and changes in policy that in olve
the text portion of Subdivision 0 are discussed in the body of the “Overview” under the appropriate
study (e g. what material to feed in livestock metabolism studies, when are processing studies needed)
Page 4 Second to last paragraph, last line
Change ‘other use relevant use limitations”
To “other relevant use limitations”
Page 7 Second paragraph, first line
Change ‘For domestic pesticides uses’
To. “Foi drmestic pesticide uses’
Page 8 Second paiagiaph, line four
Change ‘in which pesticide are applied’
To “in which pesticides are applied”
Third paragraph, first line
Change ‘Clearly written and practical uses”
To “Clearly written and practical use”
Page 18 Second paragraph, line 15
ChangeS ‘samples of these crop should”
To “samples of these crops should’
Page 19 Third paragraph, line four
Change: “Vol. 24
To. “Vol. 29”
Page 21 Last two lines
Delete all words following ‘data indicate that”
Page 23 Third paragraph, last line
Change. ‘part (b)’
To: “part (ii)”
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December 24, 19? 9
PHASE 3 TECHNICAL GUIDANCE
Page 24 Last paragraph. first line
Change ‘When the use of pesticides in agricultural buildings are such”
To “When the use of pesticides in agricultural buildings is such”
Page 30 Third paragraph, line 8
Change “it may be appropriate to propose combined tolerance for”
To “it may be appropriate to propose a combined tolerance for”
Page 32 First paragraph. lines 12 and 13
Change’ “toxicologically-innocuous, then additional data will be required In some case”
To “toxicologically innocuous, then additional data will be required. In some cases”
Page 34 Paragraph (2)
Change “(2) Pyrolysis products derived from the active ingredient must be characterized.’
To “(2) Total residues on cured or dried tobacco. If residues at 0 1 ppm or more are
detected, the determinations in the next paragraph are required.
(3) Pyrolysis products derived from the active ingredient must be characterized and the
level of residue in smoke must be quantified”
Page 35 First paragraph
Replace the first paragraph with the following 2 paragraphs.
“Uses on crops grown for seed only are considered to be non-food uses if (1) subsequent to
treatment the crop is not fit for consumption by humans and is not fed to livestock, and (2) there is no
likelihood of residues in crops grown from the harvested seed. Otherwise, such uses are considered to
be food uses requiring tolerances.
‘Examples of uses where the crop is considered unfit for consumption subsequent to treatment are
post-bloom applications to leafy vegetables and Brassica (cole) leafy vegetables, and desiccant uses on
okra and root and tuber vegetables These uses would qualify as non-food uses provided there is no
likelihood of residues in crops grown from the harvested seeds Factors affecting this include the level
of residues on the harvested seed, the half-life of seed residues, the weight of the seed in relation to
that of the subsequent crop, and the amount of residue uptake into the aerial portion of the crop’
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Deccmber 24, lt)
PHASE 3 TECHNICAL GUIDANCE
ATTACHMENT 2
Pesticide Assessment Guidelines
Subdivision 0
Residue Chemistry
REVISIONS EN TABLE II
This Attachment lists the revisions that have been made in Table II of the Residue Chemistry
Guidelines since 1982. This list includes both typing errors from the original document as well as
changes such as new feed items (e g, potato waste) and elimination of practical feeding restrictions (e g,
sugar beet tops). Typing errors in the text portion of the original Subdivision 0 are shown in
ATTACHMENT 1 to this “Overview’ Guideline clarifications and changes in policy that involve the
text portion of Subdivision 0 are discussed in the body of the ‘Overview” under the appropriate study
Addition of Grain Dust
Since this revision affects numerous crops, it will be listed here for emphasis and convenience
‘Grain dust’ should be added under ‘PROCESSED COMMODITIES’ and “FEEDS” for the
following crops barley (page 41), buckwheat (page 42), corn, field (page 44), millet (page 49), oats
(page 50), rice (page 53), rye (page 53), sorghum. grain (page 54), soybeans (page 54) and wheat
(page 57). With regard to the %‘s of the livestock diets, the figure “20’ should be placed under all
types of animals in Table II. Data on grain dust from wheat will cover barley, buckwheat, millet,
oats, and rye. The other crops will each need their own study. Also, as described on page 4 of the
Overview, grain dust data are needed only when detectable, primarily surface residues are present on
the grain. IThis requirement was instituted due to the increased usage of grain dust in animal feed
as a result of laws restricting its disposal I
Page 40
Alfalfa
Change ‘Forage(G) ” to “Forage(G) ’ and “NU 3 ” to ‘NUn” [ These footnotes were inad e’tently
reversed in the original published guidelines.
Page 4!
Barley
Delete “Hay’ from the “RAC” and “FEEDS” columns and all its corresponding numbers across the
chart Delete ‘Germ’ and “Milling products” in the ‘PROCESSED COMMODITIES’ column and
replace with the terms ‘Hulls, Bran, Flour, Pearl Barley’. Also, add “Grain dust’ as specified above.
[ ‘Hay” was deleted since it is not a significant feed item. “Milling products’ were defined to clarify
which commodities need to be analyzedj
Beet, garden
Change “Greens” to “Leaves’ in the “RAC” column. (The term was changed to clarify the part of
the plant that makes up the RAC.J
Page 42
Buckwheat
Delete ‘Bran” and ‘Milled products’ in the ‘PROCESSED COMMODITIES’ column and replace
with the terms ‘Hulls, Middlings, Flour’. Also, add ‘Grain dust’ as specified on page 1 of this
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December 24. 1989
PHASE 3 TECHNICAL GUIDANCE
ATTACHMENT [ “Milled products” were defined o clarify which commodities need to be
anaI zed
Page 43
Chicory
Add the term “Roots” under the “RAC” column [ “Roots” was inadvertently omitted from the
published document
Citrus
Delete ‘Peel’ under “PROCESSED COMMODITIES”. (Citrus peel is not a significant food or feed
item
Page 44
Corn, field
Delete all the terms under ‘PROCESSED COMMODITIES” and replace with the followingS
“Wet Milling
Starch
Crude oil
Refined oil
Dr Milling
Grits
Meal
Flour
Crude oil
Refined t il’
Also, add “Grain dust” as specified on page 1 of this ATTACHMENT. In the row ‘Silage” under
the columns ‘BEEF’ and “DAIRY”, change the numbers “25 10’ to “30 50”. [ “Milled products”
were defined to Uarify which commodities need to be analyzed Several percent of the diet figures
in the published guidelines were incorrect.)
Corn, pop
In the row ‘Fodder(G)” under the columns “BEEF’ and “DAIRY”, change the numbers “3() 5fl’ to
“25 10’ [ These figures were incorrect in the published guidelines
Corn, sweet
Delete the “(G)’ following “Forage” in the “FEEDS’ column. In the row ‘Cannery waste” under the
columns “BEEF’ and “DAIRY”, change the numbers “30 50” to “25 10’ [ With the possible
exception of uses limited to Florida, we do not consider a feeding restriction on sweet corn forage
to be practical. The percent of the diet figures in the published guidelines were incorrect.)
Page 45
Dates
Under the column ‘PROCESSED coMMoDmEs” add the term “Dried”. [ This commodity was
inadvertently omitted from the published guidelines.)
Page 46
Flax
Delete “Hulls” from both the “PROCESSED COMMODITIES” and “FEEDS” columns and all its
corresponding numbers across the chart. [ Flax hulls are no longer considered a significant feed
item.)
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December 24. l9S9
PHASE 3 TECHNICAL GUIDANCE
Page 47
Guar
Delete “Milled products” and replace with “Meal” and “Gum”. [ “Milled products” were defined to
clarify which commodities need to be analvzed.J
Page 48
Lentils
Add “(G) ” after both “Forage” and “Hay” in the “FEEDS” column. (We have concluded that a
feeding restriction for these commodities is practical.I
Page 49
Millet
Delete “Milled products” under “PROCESSED COMMODITIES” and replace with “Hulls, Flour,
Meal”. Also, add “Grain dust” as specified on page 1 of this ATTACHMENT [ “Milled products”
were defined to clarify which commodities need to be analyzed.1
Mint
Add “(G)” after “Spent hay in the “FEEDS” column. [ We have concluded that a feeding restriction
for this commodity is practical.)
Mustard
Delete “Seeds” from the “PROCESSED COMMOD1TIES column and add it to the “RAC” column.
[ Since mustard seed is harvested in the field, it is a raw agricultural commodity and not a processed
commodity
Page 50
Oats
Delete ‘Has” and Hulls” under the “RAC” column. Also delete “Hay(G)” under “FEEDS” and its
corresponding numbers across the chart. Under “PROCESSED COMMODITIES’ delete “Milled
products” and replace with “Hulls, Flour, Rolled Oats” Also, add “Grain dust” as specified on page
I of this ATTACHMENT. [ Oat hay is not a significant feed item. “Milled products” were defined
to clarify which commodities need to be analyzed I
Page 51
Peppermint
Add “(G)” after “Spent hay” in the “FEEDS” column. [ We have concluded that a feeding restnction
for this commodity is practical.)
Page 52
Potato
Delete “Dned” under “PROCESSED COMMODITIES” and add “Wet peel” and “Dry peel”. Add
“Processed potato waste’ under ‘FEEDS’ and the following percents of livestock diet: BEEF.50,
DAIRY.25, TURKEY BROILERS.NU, LAYING HENS-lO, SOWSIBOARS-50, FINISHING
SWINE-SO. Feed additive tolerances for processed potato waste should be based on the maximum
concentration factor observed for residues in/on granules, wet peel, or dry peel. [ Potato waste has
been determined to be a significant livestock feed item.)
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PHASE 3 TECHNICAL GUIDA NCE
Page 53
Rape
Under ‘PROCESSE COMMODITIES” replace “Oil” with “Crude oil” and Relined oil” [ This
makes the requirements for rape seed oil consistent with those of other vegetable oils I
Rice
Delete “Milled products’ under ‘PROCESSED COMMODITIES” and replace with “Bran” Under
‘FEEDS” delete the “(G)’ after ‘Straw’ Also, add ‘Grain dust” as specified on page 1 of this
ATTACHMENT [ It has been determined that a feeding restriction for rice straw is not practical
“Milled products’ were defined to clarify which commodities need to be analyzed I
Rye
Delete “Milled products” under “PROCESSED COMMODITiES” and replace with ‘Flour, Rye feed
(mix of bran, red dog flour, and middlings)”. Also, add “Grain dust’ as specified on page 1 of this
ATTACHMENT. [ ‘Milled products” were defined to clarify which commodities need to be
analvzed.I
Safflower
Under ‘PROCESSED COMMODITIES’ replace “Oil’ with ‘Crude oil’ and ‘Refined oil” [ This
makes the requirements for safflower oil consistent with those of other vegetable oils I
Page 54
Sorghum. grain
Replace ‘Milled pdts’ with “Starch” under ‘PROCESSED COMMODITIES”. Also, add “Grain dust’
as specified on page 1 of this ATTACHMENT. [ This is to clarify which commodities need to be
analyzed. I
So heans
Under “RAC” and “FEEDS” delete “Straw’ and its corresponding numbers across the chart Also.
add “Grain dust” as specified on page 1 of this ATTACHMENT [ Soybean straw is not a significant
feed item.j
Spearmint
Add “(0)” after ‘Spent hay’ under the “FEEDS’ column [ It has been determined that a feeding
restriction is practical for this commodity.
Page 55
Sugar beet
Delete the “(0)” after “Leaves’ under “FEEDS’. [ A feeding restriction for sugar beet leaves or tops
has been determined to be not practical.]
Add the following three crops after ‘Sunflower” on page 55.
CROP RAC
“Sweet potato (yam) Root
Swiss chard Fresh
Taro Root
Foliage’
[ These crops were omitted in the published guidelines I
Page 56
Tomato
Under “PROCESSED COMMODmES’ replace “Puree” and ‘Catsup’ with ‘Puree or Catsup”.
[ Data on one product suffices for the other due to similar water conicnt.1
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December 4 l9 N
PHASE 3 TECHNICAL GUIDANCE
PaEe 57
Wheat
Under “RAC” delete “Hay” and under “FEEDS” delete “l-Iav(G)” and all as corresponding numbers
across the chart. Delete “Milled products” under “PROCESSED COMMODITIES” and replace with
“Bran, Flour, Middlings, Shorts”. Delete the “(G)” after “Straw” under the “FEEDS” column Also.
add “Grain dust” as specified on page 1 of this ATTACHMENT [ Wheat hay is not a significant
feed item “Milled products” were defined to clarify which commodities need to be analyzed Since
wheat straw is a major by.product and is fed in times when roughage is in short supply, the grower
restriction for wheat is not appropriate
FoOtnote (3
Revise t(, i-Under grower control and not routinely fed to livestock, therefore, subject to label
restrictio tgainst feeding” [ This revision clarifies that there are two factors to consider in the
practicali )f feeding restrictions. First, the feed item needs to be in control of the grower (i e.
does not leave the farm). Second, the feed item can not be of such importance that it will be
routinely fed regardless of label restrictions. For example, alfalfa hay is a valuable animal feed and
is virtually always raised for that purpose. Therefore, a label statement prohibiting feed use of
alfalfa hay would often be ignored I
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December 24. 1989
PHASE 3 TECHNICAL GUIDANCE
ATTACHMENT 3
July 25. 1989
MEMORANDUM
SUBJECT’: Guidance on When and i-low to Conduct Livestock Metabolism Studies.
FROM Richard D. Schmitt, PhD, Chief
Dietary Exposure Branch
Health Effects Division (l-17509C)
TO Dietary Exposure Branch Staff
This memo is intended to clarify when livestock metabolism studies are needed and the efforts that
should be made to identify residues in such studies The emphasis will be on those cases where low or
non-detectable residues are observed in feed items. The need for conventional livestock feeding studies
in such situations will also be addressed
For many years Branch policy called for livestock metabolism studies only when detectable residues
of concern were tound in feed items in crop field trials. This procedure discnminateS against registrants
who develop more sensitive methods for measuring residues in crops In recent times we have requested
animal metabolism studies in some instances when no detectable residues are present in feed items
However, this procedure has not been consistently applied. Effective with this memorandum, it will be
Branch policy to require livestock (ruminant and/or poultry) metabolism studies whenever a pesticide is
to be applied to a crop having an animal feed listed in Table II of the Residue Chemistry Guidelines.
In addition to not discouraging the development of more sensitive methods for crop residues, this policy
will provide some information on the potential transfer of residues to meat and milk in those cases
where misuse results in residues in feed items not expected to have residues from approved uses.
Since ruminant and poultry metabolism studies are now to be required in cases where very low or
non-detectable residues are present on feed items, the question arises as to what dose level is
appropriate. Following FAQ’s lead, we suggest =10 ppm in the diet as the dosing level. This will
typically represent 200-l000x the anticipated dietary burden for livestock when trace residues occur in
feed items. The radiolabeled material should have sufficient specific activity to enable detection of =001
ppm (10 ppb) in milk, tissues, and eggs.
As already stated in the Guidelines, the ruminants and/or poultry (depending on feed items
involved) should be dosed with radiolabeled matenal for at least three consecutive days with milk or
eggs collected daily Animals should be sacrificed within 24 hours of the final dose and total
radioactivity measured in edible tissues and milk/eggs.
With regard to the degree of characterization of residues in animal (and plant) metabolism studies,
a strategy developed by Ciba Geigy may be used as a guide to what effort should be expended
(Reference 1) If total activity in a tissue (or crop part) is =0.01 ppm (1O ppb) or less, no
differentiation of the radioactivity would be required. For activity greater than =0.01 ppm the sample
should be extracted with organic solvents (perhaps mixed with neutral water). The levels of extractable
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DeLcmh r 2 - I jQ Y
PHASE 3 TECHNICAL GUIDANCE
and non-extractable (NE) activity should then be quantitated to determine the degree of characterizatiOn
that is needed If the extractable activity represents 00I ppm or less in the original sample, it need
not be examined further. For extractable activity of 001-005 ppm, the partitioning beha ior between
aqueous and an organic solvent should be determined followed by chromatOgraphic (TLC, HPLC)
analysis of the organosoluble activity. The chromatographic behavior of this activity can be compared to
that of the parent pesticide and likely nietabolites. When the extractable activity exceeds =005 ppm.
complete characterization should usually be attempted (however, note discussion on highly exaggerated
doses below) Identities of metabolites should be confirmed with a second technique, spectroscopic if
possible
With regard to the non-extractable (NE) residues obtained above, further characterization should be
attempted when the NE activity in a sample is greater than =0 1 ppm or 10% of the total activity in
that sample. Attempts should be made to release these residues by acidic or basic hydrolyses and/or
enzyme treatments The degree of characterization needed for the released activity will depend upon its
total level in ppm using the figures outlined in the previous paragraph (ie, =0.01,001-005, and >005
ppm) for extractable residues.
It should be noted that the above ppm “trigger” values are not absolute requirements but rough
guides as to how much characterization is adequate. En the metabolism studies wherein highly
exaggerated feeding levels are employed and low activity results in tissues, characterization requirements
should be less stringent than when expected dietary burdens lead to significant activity in animal
products. For example, if the anticipated dietary burden to livestock is about 0.0! ppm, 10 ppm radio-
labeled compound is fed (I000x), and total activities in tissues, milk or eggs are 0.1 ppm, minimal
characterization of residues should be adequate (unless toxicologists express a special concern with
residues at this level Such situations often arise with early season herbicides having low application
rates.
On the other hand, when activities 0 I ppm are observed in animal commodities from ingestion of
the pesticide at levels expected on feed items, thorough identification of the residues is generally
required. This is likely when pesticides are applied foliarly at high rates through the entire growing
season
Finally, with iespect to the need for conventional feeding studies, such data will not be required
when no detectable residues are observed in feed items from crop field trials reflecting the proposed use
of the pesticide (maximum rate, minimum preharvest interval) unless the metabolism study shows
potential for significant bioaccumulatiOn. When trace residues are detected in the field tnals, the
reviewer should consider the anticipated dietary burdens and the results of the radiolabeled metabolism
study when determining whether feeding studies are necessary. In the previously cited example (0.01
ppm dietary burden, l000x dose leading to <0.1 ppm total activity in meat/milk/eggs), a feeding study
would not be necessary as expected residues in animal commodities from ingestion of 0.01 ppm would
be on the order of =0.1 pp (assuming linear relation-ship between dose and residues). In this case the
metabolism study also serves as a feeding study and tolerances would not be needed for meat, milk,
poultry and eggs.
SUMMARY
sLivestock (ruminant and/or poultry) metabolism studies will be required whenever a pesticide is to be
used on a crop having a livestock feed item in Table hot the Residue (lemistry Guidelines.
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PHASE 3 TECHNICAL GLIDANCE
.Tngger valu& (ppm radioactivity) are provided as rough guides to the degree of characterization
needed in metabolism studies. However, reviewers should take into a unt how the rate or d e used
in the metabolism study compares to that expected from actual use of the pesticide.
• When considering the need for conventional feeding studies in scs where low residues are found in
feed items, reviewers should take into account the expected dietary burdens and results of animal
metabolism studies. The lauer may indk te that residues in animal commodities uld be well below
detection limits and thus serve as feeding studies. Meatimilk/egg tolerances uId not be necessary in
such situations.
Reference I “STRATEGY FOR DETERMINATION OF EXTENT OF METABOLISM STUDIES
AND DEVELOPMENT OF RESIDUE METhODS BASED ON TRIGGER VALUES’, 1127/88, Dr B
Donzel. Ciba-Geigy.
cc. RF, Petition Review Aids File
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DcLemhcr 24 l - )
PHASE 3 TECHNICAL GUIDANCE
ATTACHMENT 4
SUGGESTED STATES FOR CROP FIEIJ) TRIALS
The following lists were prepared in 1983 in response to an IR-4 request as to which states should
be included in field trials for representative crops to obtain crop group tolerances. Field trial sites
should be representative of the area(s) in which the crop is grown in each state and of the prevailing
cultural practices Arid regions employing furrow irrigation (especially California) should be included
for most crops since higher residues are often found under such conditions. Seasonal variation should
also be represented.
In cases where a slash appears between states, any one of the states could be used as a site for the
residue trial. If a comma appears between states, even if they are in the same region, data for both
states is recommended due to the importance of the food crop (e.g., potatoes versus radishes). Residue
data for a state/area should consist of a composite sample from several fields. Of course if a state such
as PA is recommended for residue trials, substitution of an adjacent state such as NY would be
considered acceptable as long as the total number of trials remains the same.
With regard to obtaining tolerances on individual crops (as opposed to crop group tolerances), the
lists of states below would suffice in most cases wherein no detectable or low residues are consistently
observed in the trials. This would typically be the case for early season, low rate applications of
herbicides It should be noted, however, that for those crops where only I or 2 states are shown (c.g.,
almonds, garlic, radishes) two or more locations should be included in each state.
For those uses which result in significant residues on food or feed portions of crops, the conducting
of just one trial in each of the states would usually not be adequate to obtain a tolerance on an
indi idual crop. Either multiple locations in each state or the addition of more states should be
considered in such instances. Foliar, high rate applications of fungicides or insecticides throughout the
entire growing season would usually fall under this category.
Bulb vegetables (ailiu
Garlic CA
Green onion TX, CA, AZJNM
Bulb onion NY, MI, OR/WA, ID, CO
Roots and tuber vegetables (and their leaves )
Carrots CA, TX/AZ, MI/WI, WA/OR, NY/NJ, OH, MN
Potato ID, OR/WA, ND, MN, WI, ME, CA, CO
Radish FLCA
Sugarbeets CA, MN/ND, ID, WA, NE, WY, MI
Roots and tops
Turnip CA. OR/WA. TX, OH, NJ, ILJIN
Sugarbeets See Sugarbeets above
Leafy vegetables (non-Brassica )
Lettuce (head and leaf) CA, FL, TX/AR, NY/NJ, CO, WA
Celery CA, FL, MI, WA
E-28
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DcLcrnhcr -.
PHASE 3 TECH\ICAL GLID k\CE
Spinach CA. TX1OK. NJ. MD/VA CO/AR
Brassica egetables
Broccoli CA TXJAZ. OR
Cabbage NY, CA. FL, TX. WI. NJ. SC/NC/GATN
Mustard Greens CA, TXJAZ. MI/OH/IN, FL LA/GAjTN
Legumes (succulent or dried )
Snap beans NJ,NY, TN/NC/VA. CA. Ml. FL
Green peas CA DE, ID, MN, WI, OR/WA
Dry beans CA ID. MI, CO. NE. ND
Dry peas WAJOR. ID
Soybeans MS/LA TN, AR, IN/IL, 10/NE, MN, MO
Legumes (foliage )
Beans See “Legumes above
Field peas See “Legumes above
So beans See Legumes ” above
Fruiting Vegetables
Tomato CA, FL, OH/PA. NJ, IN, MI, SC/TN
Peppers CA. FL TX, NC, NJ
Cucurbits
Cantaloupe CA. TX/AZ, IN, MI, GA/SC
Summer squash CA, FL, TX, NJ/MA/NY, OR, GA/SC, MI
Cucumber CA, FL, TX, Ml, NY/NJ, NC/SC. OH
Citrus
Orange FL, CA. AZ/TX
Grapefruit FL, TX, CA
Lemon CA. AZ
Pome Fruits
Apples CA MI, NY, PAIWV, VA/NC, WA/OR
Pears CA, MI, NY, WA
Stone fruits
Peach CA, GA/SC, MI, NJ/PA WA
Cherry CA, OR/WA, MI, UT/MT/ID, NY/PA
Plums CA ID, MI, OR/WA
Small fruits and berries
Blackberry or other Rubus spp. WA, OR
Blueberries MI, NJ, ME, NC, WA/OR
Strawberry CA FL, OR/WA, IN/MI, NY/OH, LA
Cranberry WI, MA/NJ, OR/WA
Grape CA, NY, WA, Ml, NC
E.29
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DcLcmher 4 l J
PHASE 3 TECH\ICAL GLIDA. CE
Tree nuts
Almond CA
English walnut CA, OR
Pecans AL/GA’LA/MS, NMID(IOK
Cereal grains
Wheat All areas across the country
Sorghum MO/KS, IL, TX/OK/NM, SD, NE, NC/GA. CO
Rice AR, CA, LA, TX, MO
Corn (sweet) FL, CA, NY, TX. OH/PA. MAJNJ, OR/WA’ID, MI/MN/WI, IL
Corn (field) All areas across the country
Forage, fodder and straw of cereal grains
Corn See Corn’ above
Wheat See “Wheat’ above
I other cereal grain crop See “Rice” above
Grass (including fodder and hay
Bermuda All areas across the country
Blue
Brome
Fescue
Non-grass animal keds (forage, fodder, straw and hay )
Alfalfa All areas across the country
Clover
Herbs and spices
Basil CA
Chives CA
Marjoram CA
Sage CA
Dill Southern state, Western state
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Dcumher 4, l Y
PHASE 3 TECHNICAL GUIDANCE
APPENDEX P ENSTRUC11ONS FOR ELECTRONiC REPORT [ NG
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December 24. 19S9
PHASE 3 TECHNICAL GUIDANCE
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DeLember 24. 1989
PHASE 3 TECHNICAL GUIDANCE
ELECrRONEC SUBMISSION OF PHASE 3 RESPONSES
The agency is currently assessing the advantages of electronic transmission of information required under
FIFRA and its amendments We are conducting research into the many areas where this technology
would produce the highest return for both industry and government We have conducted a pilot project
with the chemical industry and determined that there are many other areas we should explore If your
company is interested in electronic reporting. please request an information packet by writing to
Electronic Reporting
Reregistration Phase 3 Response
Office of Pesticide Programs
P0 Box 14890
Silver Spring, MD 20910-4890
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December 24, 1989
PHASE 3 TECHNICAL GUIDANCE
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Dcu rnbcr 4 19
PHASE 3 TECHNICAL GUIDANCE
APPENDIX G: BLANK FORMS FOR PHASE 3 SUBMiSSIONS
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Dccembcr 24. IYSY
PHASE 3 TECHNICAL GL’IDA CE
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United States Environmental Protection Agency
EII Washington, DC 20460
PHASE 3 IDENTIFICATION OF
Form Approved
0MB No. 2070-0106
UNREASONABLE ADVERSE INFORMATION
NOT INDICATED ON THE PHASE 3 WORKSHEET
Approval expires 12-92
The public reporting burden for completing this form only is estimated to average approximately 30
minutes per response, including time for reviewing instructions, searching existing data sources,
gathering and maintaining the data needed, and completing and reviewing the collection of rnforma-
tion EPA has accounted for this burden within the total average burden associated with the collection
of information under section 4(e)(1 )(E) Send comments regarding the burden estimate or any other
aspect of this collection of information, including suggesting for reducing this burden, to Chief,
lnformationPolicyBranch ,I’M-223 U S EnvironmentalProtectionAgency ,401 MSt ,S W ,Washirig-
ton, D.C. 2c460; and to the Office of Management and Budget, Paperwork Reduction Project (2070-
0106), Washington, D.C. 20503
NSTRUCTIONS
You must use this form to identify each additional item of information on adverse effects that ‘o ,th
identified on our ‘Phase 3 Chemical Response Worksheet.” For example, you must use this torm
if you determine that you must identify adverse effects information found in studies that are not listed
in the ‘Phase 3 Chemical Response Worksheet” or if you must identify information found in incident
reports
Please note that this form does not tell you what information constitutes the adverse information that
must be identified to EPA. You must consult the guidance on identifying unreasonable ad verse ettects
information that appears in chapter 4 of the “Accelerated Reregistration Phase 3 Technical Guidance”
to determine what information must be identified. Once you have determined what information is
required, and have determined that you can not identify an item of information on the “Phase 3
Chemical Response Worksheet,” you must complete this form to identify that item
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Lnited States Environmental Protection Agency
Washington, D C. 20460
PHASE 3 IDENTIFICATION OF
UNREASONABLE ADVERSE INFORMATION
NOT INDICATED ON THE PHASE 3 WORKSHEET
Form Approved
O .{BNo 20700106
Approval expires 2
Instructions: Please type or print in ink Please read carefully the attached instructions and supply the
inforrrtation requested on this form Use additional sheets(s) if necessary.
FILL OUT THIS FORM FOR EVERY SEPARATE ITEM OF ADVERSE EFFECTS INFORMATION.
CASE
COMPANY NUMBER
NAME CHEMICAL
NUMBER
COMPANY CHEMICAL
NUMBER NAME
1. Does the information pertain to only some of your products? Yes No
if yes, list the product number(s)
2. Is the source of information a study not identified on the Phase 3 Chemical Response Worksheet, or an
incident report or other information source?
If the source is a study not on your Worksheet, fill out
section 3 below,
If the source is an incident report or other source,
fill out section 4 below.
3. A. Has the study been submitted to EPA?
Yes MRfl) Number
Date Submitted
Title
No If no, you are required to submit
a copy of the study under
section 6(a)(2) of FIFRA.
B. Attach summary of adverse effects
information contained in the study and
explain why it may indicate adverse
effects.
4. A. Has the information been submitted to EPA?
Yes Date Submitted
How and to whom
Submitted
No If no, you are required to submit
a copy of the study under
section 6(a)(2) of FIFRA.
B. Attach summary of adverse effects information
1. Identifies the source of information (e.g. legal
proceedings, letter from customer, factory report).
2. Desaibes the adverse effects and explains why
it may indicate adverse effects.
5. Certification
I certify that the statements that I have made on this form and all
attachments therein are true, accurate, and complete. I acknowledge that
any knowingly false or misleading statement may be punishable by fine or
imprisonment or both under applicable law.
5 A. Signature of Company’s Authorized Representative
6. Contact (Typed Name)
7. Phone
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United States Environmental Protection Agency
Washington, DC 20460
EPP PHASE 3 IDENTIFICATION OF
UNREASONABLE ADVERSE INFORMATION
NOT INDICATED ON THE PHASE 3 WORKSHEET
Form Approved
0MB No 2070-0106
Approval expires 12-92
The public reporting burden for completing this form only is estimated to average approximately 30
minutes per response. including time for reviewing instructions, searching existing data sources,
gathering and maintaining the data needed, and completing and reviewrng the collection of infornia-
tion EPA has accounted for this burden within the total average burden associated with the collection
of information under section 4(e)(1 XE) Send comments regarding the burden estimate or any other
aspect of this collection of information, including suggesting for reducing this burden, to Chief,
Information Policy Branch, PM-223, U.S Environmental Protection Agency, 401 M St., S.W , Washing-
ton, D C. 20460; and to the Office of Management and Budget, Paperwork Reduction Project (2070-
0106), Washington, D C 20503.
INSTRUCTIONS
You must use this form to identify each additional itemof information on adverse effects that you have
identified on our ‘Phase 3 Chemical Response Worksheet.” For example, you must use this form
if you determine that you must identify adverse effects information found in studies that are not listed
in the “Phase 3 Chemical Response Worksheet” or if you must identify information found in incident
reports.
Please note that this form does not tell you what information constitutes the adverse information that
must be identified to EPA. You must consult the guidance on identifying unreasonable adverse effects
information that appears in chapter 4 of the “Accelerated Reregistration Phase 3 Technical Guidance”
to determine what information must be identified Once you have determined what information is
required, and have determined that you can not identify an item of information on the “Phase 3
Chemical Response Worksheet,” you must complete this form to identify that item.
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United States Environmental Protection Agency
Washington. D C 20460
PHASE 3 IDENTIFICATION OF
UNREASONABLE ADVERSE INFORMATION
Form .ppro’.ed
0MB No. 70.Oi06
NOT INDICATED ON ThE PHASE 3 WORKSHEET
Approval expires 12.c
Instructions: Please type or print in ink. Please read carefully the attached instructions and supply the
information requested on this form Use additional sheets(s) if necessary.
FILL OUT THIS FORM FOR EVERY SEPARATE ITEM OF ADVERSE EFFECTS INFORMATION.
CASE
COMPANY NUMBER —
NAME CHEMICAL
NUMBER
COMPANY CHEMICAL
NUMBER NAME
1. Does the information pertain to only some of your produc ? Yes No
If yes, list the product number(s)
2. Is the source of information a study not identified on the Phase 3 Chemical Response Worksheet, or an
incident report or other information source?
If the source is a study not on your Worksheet, fill out
If the source is an incident report or other source,
section 3 below,
flU out section 4 below.
4. A. Has the information been submitted to EPA?
Yes Date Submitted
How and to whom
Submitted
No If no, you are required to submit
a copy of the study under
3. A. Has the study been submitted to EPA?
Yes MRID Number
Date Submitted
Title
No If no, you are required to submit
a copy of the study under
section 6(a)(2) of FIFRA.
section 6(a)(2)
B. Attach summary of adverse ff
information contained in the study and
B. Attach summaxy of adverse effects information
explain why it may indicate adverse
1. Identifies the source of (e.g. legal
proceedings, letter customer, factory report).
2. Desa’ibes the adverse effects and explains why
it may indicate adverse effects.
5. Certification
I certify that the statements that I have made on this form and all
attachments therein are true, accurate, and complete. I acknowledge that
any knowingly false or misleading statement may be punishable by fine or’
imprisonment or both under applicable law.
5 A. Signature of Company’s Authorized RepresentatIve
6. Contact (Typed Name)
7. Phone
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Dc cmr cr - i)
PHASE 3 TECHNICAL GLIDA.\CE
APPENDD( H: ADDmONAL GUIDANCE ON RAW DATA
EPA has developed the following lists to provide registrants with additional guidance on ho to
determine whether they have access to the raw data that support the studies they are relying on for
reregistration
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December 4. 1989
PHASE 3 TECHNICAL GUIDANCE
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DLLCmt’cr 24 l”
PHASE 3 TECI-INICAL GLIDANCE
RECORDS TO BE MAINTAINED FOR SUBDEV1SIO D
The following listing covers studies for submission under Subdivision D Records to be maintained
include but are not necessarily limited to the following
1 Name and address of testing facility and Principal Investigator
2 Name and address of registrant / test sponsor
*3 Test substance type, source, strength. identification and purity (batch/lot numbers), and, procedures
used to manufacture or synthesize the test substance
‘4 Analytical test data for all components
*5 Analytical methods
‘6 Documentation of certified limit determinations
‘7 Tesi method identification (reference) or description
‘8 Analytical raw data for test
‘9 Signed final report
10 Test substance receipt and distribution
11 List of participants and their training and experience
12 Inspection / calibration / standardization of equipment
13 Standard Operating Procedures
14. Correspondence
Minimum requirements
H.1
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DcLembcr 24. [ 9 Y
PHASE 3 TECHNICAL GUIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION E
The following listing covers studies for submission under Subdivision E in general, including
laboratory, simulaied or actual field testings as appropriate Records to be maintained include but
are not necessarily limited to the following
* 1 Name and address of testing facility and Pnncipal Investigator
‘2 Name and address of registrant / test sponsor
‘3 Test substance
- identification, strength, purity and composition
- storage requirements
- solubi litv
- storage stability
- preparation of dosages
- application procedures
- amount administered I dosage level
- stability in mixture
• solubility in mixture
‘4. Description of test site
• location
- layout (terrain, soil type)
‘5. Methodology! standard or EPA approved procedures
‘6 Environmental condition
- water depth, pH, temperature, relative humidity, salinity, dissolved oxygen, etc.
‘7. Census of existing population
- mammalian, avian, amphibians, insects, etc.
• vegetation, soil types, etc.
• mobile species (transients): lish, wildlife, humans, etc.
‘8. Set up of study
number of animals
size, age, sex
* Minimum requirements
H-2
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Deccmbcr 24. l9S9
PHASE 3 TECHNICAL GUIDANCE
SUBDIVISION E CONTINUE!)
‘9 E.xperimental data
- weights
• mortality
- observations/behavior
‘10 Sample collection data
• 11 Activities around field / test site and unforeseen circumstances that may affect test results
12 Signed final report
13 Test substance receipt and distribution
14 Animal receipt, randomization, distribution
15 Animal health records prior to the start of the study
16 List of participants and their training and experience
17 Inspection / Lalibration / standardization of equipment
18 Standard Operating Procedures
19 Correspondence
20 Housing conditions: number per container, lighting, etc
21 Method of counting
• Minimum requirements
H-3
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December 24. 19S9
PHASE 3 TECHNICAL GUIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION F
The following listing covers studies for submission under Subdivision F in general, including
mammalian toxicity and mutagenicity testings as appropriate Records to be maintained include but
are not necessarily limited to the following
‘1 Name and address of testing facility and Principal Investigator
*2 Name and address of registrant / test sponsor
‘3 Test substance
- idenuflcation, strength. purity and composition
- solubility
- storage requirements
- storage stability I viability of the cell cultures
- preparation of dosages
- amount administered I dosage level
- application procedures
- stability in mixture
- solutility in mixture
*4 Methodology / standard or EPA approved procedures
‘5 Environmental condition
‘6 Set up of study
- number of animals
- size. age, sex
- cell culture (test system)
- type of media used
‘7 Experimental data
- weights
• mortality / viability of cells
- observations / behavior
• feed consumption
- analytical chemistry
• homogeneity of dosage preparation
• stability of dosage preparation
• dosage analyses
- genotoxic effects
- clinical patholo r
* Minimum requirements
H -4
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DeLLmt’Lr 24 1mY
PHASE 3 TECHr\ICAL GLlDA CE
SUBDIV!SION4 F CONTINIJED
- necropsy
• wet tissues
• histopatholo ’
- tissue blocks I slides
‘8 Unforeseen circumstances that may have an effect on the test results
‘9 Signed flnal report
10 Test substance receipt and distribution
11 Animal receipt, randomization, distribution
12 Animal health records prior to the start of the study
13. List of participants and their training and experience
14 Inspection I calibration I standardization of equipment
15 Standard Oper.iting Procedures
16 Correspondence
* Minimum requirements
H-5
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DeLember 24. i9? 9
PHASE 3 TECHNICAL GU1DA.NCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION I
This general listing covers studies for submission under Subdivision J, including greenhouse or field
tests as appropriate Records to be maintained include but are not limited to the foIlo ing
‘1 Name and address of testing facility and Principal ln esugator
‘2 Name and address of registrant I test sponsor
s3 Test substance
• chemical name, percent active ingredient and formulation, amounts of impurities. ir
solvents. etc
- water solubilitv
• vapor pressure
- solvents in controls
‘4 Date of test initiation and termination
s5 Test organism
- hisors and source
- identification of test species (family, genus, species, cultivar / variety)
‘6 Test design
- observation period
- criteria used to measure effects
• test solution, preparation method
- water source and description
- test vessel size, type
- dose rates in ppm and #aiJA, number of treatment levels
- type and number of controls
- number of replicates
• physical and chemical properties (pH, % organic matter) of substrate / media
- protocols used for growth measurements and rating systems
- test organism and control group assignment method
- size, location, and air flow of growth chamber, greenhouse, field plot
- environmental conditions: thermoperiod, relative humidity; lighting regime - intensity,
quality, photopenod; watering regime • frequency, amount
• cultural practices
‘7. Statistical analysis method and results
* Minimum requirements
H-6
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December 24. l9 9
PHASE 3 TECHNICAL GUIDANCE
SUBDIVISION J CONTINUED
8 Discussion and results - signed final report with
• EC25 and EC5O values in ppm and #ai/A with 95% confidence limits
- Test vs control growth measurements from which calculations are made (cells/ml/ essel.
plants/vessel, etc)
- No observed effect level
- Phytoxicity symptoms observed
9 Standard Operating Procedures, EPA protocols used
10 Deviations from EPA guidelines and rationale Report problems encountered during the study and
the effect they may have had on the study outcome
11 Inspection / calibration / standardization of equipment
• 12 Correspondence
13 Test substance receipt and distnbutiori
* Minimum requirements
H-i
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DLc mbcr 2- l’- ’
PHASE 3 TECHNICAL GUIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION K
The following listing covers studies for submission under Subdivision K. Records to be marntained
include but are not necessarily limited to the following.
‘1 Name and address of testing facility and Principal Investigator
‘2 Name and address of registrant I test sponsor
‘3 Test substance type, source, identification (batch or lot numbers) and strength
4 Identification of facility and test plots
‘5 Informed consent documentation for testing on humans, if appropriate
*6 Test system identification, source and preparation
‘7 Test substance preparations how and when test substance was prepared (diluted)
‘8 Test substance applications how and when applied and amount applied
‘9 Sampling tcUinique, identification, preservation and transfer to laboratory
‘tO All analvilLal raw data, including quality control determinations
‘11 Stability data, if appropriate, for test substance / metabolites Frozen, if analyses are more than one
week after sampling; not frozen for analyses one day after sampling
‘12. Signed final report
13 Test substance receipt and distribution
14 List of participants and their training and experience
15 Inspection / calibration / standardization of equipment
16 Standard Operating Procedures
17. Correspondence
* Minimum requirements
H-8
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Deccmoer 24 I9S.’
PHASE 3 TECHNICAL GUID \CE
RECORDS TO BE MAINTAINED FOR SUBDIVISION L
The following listing covers studies for submission under Subdivision L in general. including
laboratory and simulated or actual field tests as appropriate. Records to be maintained include but
are not necessarily limited to the following
* Test sponsor (name of study owner)
‘2 Name and address of testing facility
‘3 Name of principal investigator
‘4 Location where study was conducted
‘5 Test dates
‘6 Location of raw data and final report
‘7 Test substance
- common and trade name
- chcm;cal name and composition
• percent active ingredient
• source, lot number, or code
‘8 Preparation of test material:
- vehicle used to dissolve or dilute test substance
- preparation methods
- total amount of test material used
‘9 Test insect.s
- source of supply of test bees
- methods used to obtain test bees from colonies
• history of test bees
- health status of test bees
- age of bees at test initiation
*10 Test design:
• Size and composition of test cages
- Procedures used to prepare toicicant solution
- Criteria used to determine effects
- Amount of test material per bee or per acre
* Minimum requirements
H-9
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December 24. i9S9
PE-L SE 3 TECHNICAL GUIDANCE
SUBDIVISION L CONTINUED
- Method of administration I application
- Name of treated crop
- Weather conditions during and after application
- Method used to harvest treated foliage
- Amount of treated foliage per cage
- Rationale for selection of method, route, or frequency of application, if different from that
recommended in Subdivision L
- Method used in assigning bees to test and control groups
- Number of treated levels
- Number of replicates used
- Number of bees per treatment level and control
- Method used to determine treatment levels
- Number of bees per cage
- Number and types of controls
- Environmental conditions (ambient temperature, humidity, weather conditions)
- Source and availability of food and water
- Length of total observation period
- Frequency and duration of observations
- Length of study
11 Statistical analysis (method or references)
12 Reported results:
- LDSO in micrograms per bee, with 95% confidence limits
- No-observed-effect level
- Slope of the dose-response line, if the method provides a slope
- Percent mortality of test bees at each treatment level and time interval
- Raw mortality data
- Detailed description of all observed toxic effects
• Description of any deviation from recommended protocol
* Minimum requirements
H-IO
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DcLcmhcr 24 l 9
PHASE 3 TECHNICAL GLIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION ‘4
The following listing covers studies for submission under Subdivision M for microbial pest control
agents, including laboratory, simulated or actual field testings as appropriate Biochemical pest
control agents are tested using protocol similar to conventional chemical pesticides and, for
biochemical pesticides, the record maintenance requirements for conventional chemical testing
should be followed Records to be maintained for microbial pesticides include but are not
necessarily limited to the following’
‘1. Name and address of testing facility and Principal Investigator
‘2. Name and address of registrant / test sponsor
‘3 Test substance
a All criteria used to arrive at characterization, including taxonomic placement of the
microorganism.
b Information on the developmental biology and reproduction cycle of the microorganisms.t
c Confinement measures of the microorganisms
d Known pathogen features of the organism that may vary with the environment
e Units of active microbial ingredient in dosing I test material.
f Expected limits of detection of the microorganism.
g. Maintenance of test substance.
h. Preparation of test material for dosing.
i. Characterization of any microbial contamination
Source for each cell line / parental material
k. Survival of the microorganisms over the course of the test
1. Storage requirements (temperature)
m Application procedures
n. Stability of test substance in mixtures
t Required only for field studies
• Minimum requirements
H-Il
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December 24. 19 S9
PHASE 3 TECHNICAL GUIDAJ’ CE
SUBDIVISION M CONTINUED
4 Descripuon and location of test site.’
*5 Methodolo i / standard or EPA approved procedures
a Description of test protocols used
b. Analytical methods for cultunng, isolation, identifying, and assuring the purity of the
microbial cultures used in the experiments.
6 Environmental or testing conditions: water depth, pH, temperature. relative humidity, etc
•7 Study design
a. Size, sex, age, number, etc. of animals
b Acclimation of test system, housing
c. Animal receipt, randomization, di.stnbution
d. Animal health records prior to the start of the study
8 Census ol ex,sting populationt
a. Mammalian, avian, amphibians, insects, etc.
b. Vegetation, soil t - ‘. etc.
c. Mobile species (transients): fish, wildlife, humans)
•9 Sample collection datat
• 10. Unforeseen circumstances and activities around field / test site that may have an effect on the test
resultst
‘11 Signed final report
12. Test substance receipt and distnbution.
t Required only for field tests
* Minimum requirements
H-12
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December 24 l9 9
PHASE 3 TECHNICAL GUIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION N
The following listing covers studies for submission under Subdivision N Records to be maintained
include but are not necessarily limited to the following
* 1 Name and address of testing facility and Principal Investigator
‘2 Name and address of registrant / test sponsor
‘3 Test substance type, source, identification (batch or lot number) and purity
‘4 Test system source, identification and preparation (also processing, if appropriate)
‘5 Test locations
- Geographical location of facilities
- Specific locations of treatment plots
*6 Application of test substance to test system
- How and when test substance was prepared
- How and when test substance was applied
- Amou’it applied
- Mete’rological conditions at application
- E4uipment conditions at application
‘7 Test system sampling and sample identification
*8 Sample preser ation and shipment to laboratory
‘9 All analytical data with quality control determinations
* 10 Stability data, for test substance and I or metabolites: frozen, if analyses are more than one week
after sampling; not frozen for analyses within one day after sampling.
‘11. Signed final report
12. Test substance receipt and distnbution
13 List of participants and their training and experience
‘14 Inspection / calibration / standardization of equipment
‘15 Standard Operating Procedures
*16 Correspondence
* Minimum requirements
H-13
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December 24. 1989
PHASE 3 TECHNICAL GUIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION 0
The following listing covers studies for submission under Subdivision 0 Records to be maintained
include but are not necessarily limited to the following
* 1 Name and address of testing facility and Principal Investigator
‘2 Name and address of registrant / test sponsor
‘3 Test substance type, source, identification (batch or lot number) and strength
‘4 Test plot locations
- General geographical location of facility
- Specific locations of treatment and controls within facility
- Weather conditions at the test plot
- History of any previous pesticide use on the test plot
‘5 Test system, identification and preparation (also processing, if appropriate)
‘6 Test substance preparations and applications
- How and when test substance was prepared (diluted)
- How md when test substance was applied
- A,m’i”nt of test substance applied
‘7 Test system smmp(ing preparation, including procedures for subsampling (when and how) and sample
identification
‘8 Sample preservation and shipment to laboratory
‘9 All analstical raw data, including quality control determinations
1O Stability data, if appropriate, for test substance and I or metabolires. fTozen, if analyses are more
than one week after sampling; not frozen for analyses within one day after sampling.
‘ii. Signed final report
12. Test substance receipt and distribution
13. List of participants and their training and experience
‘14. Inspection / calibration I standardization of equipment
15. Standard Operating Procedures
16. Correspondence
* Minimum requirements
H-14
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Decemh r .- . IY 9
PHASE 3 TECHNICAL GLIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION R
The following listing covers studies for submission under Subdivision R Records to be maintained
include but are not necessarily limited to the following
1 Name and address of testing facility and Principal Investigator
*2 Name and address of registrant / test sponsor
‘3 Test substance type, source, identification (batch or lot number) and purity
‘4 Test system source and preparation
‘5 Test locations
- Geographical location of facilities
- Specific locations of treatment plots
‘6 Application of test substance to test system
- Flow iest substance was prepared
- How and when test substance was applied
- Quantity applied
• Meteorological conditions at application
- Equipment conditions at application
‘7 Test system sampling and sample identification
*8 Sample preservation and shipment to laboratory
*9 All analytical data with quality control determinations
‘10. Stability data for test substance until analyzed
‘11 Signed final report
12 Test substance receipt and distnbution
13. List of participants and their training and experience
‘14 Inspection / calibration / standardization of equipment
‘15 Standard Operating Procedures / Protocols
‘16 Correspondence
* Minimum requirements
H-15
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December 24. 1989
PHASE 3 TECHNICAL GUIDANCE
RECORDS TO BE MAINTAINED FOR SUBDIVISION U
The following listing covers studies for submission under Subdivision U Records to be maintained
include but are not necessarily limited to the following
*1 Name and address of testing facility and Principal Investigator
‘2. Name and address of registrant / test sponsor
‘3 Test substance type, source, identification (batch or lot number) and strength
‘4 Identification of facility and test plots
‘5. Informed consent documentation for testing on humans
‘6 Test system (dosimeter) preparation and locations
‘7 Test substance preparations and applications
- How and when test substance was prepared (diluted)
- Ho nd when test substance was applied
• Amc unt applied
‘8 Sample identification, preservation and transfer to laboratory
‘9 All analytical raw data, including quality control determinations
*10 Stability data, if appropriate, for test substance if analyses are more than one week after sampling
‘11 Signed final rLport
12. Test substance receipt and distribution
13. List of participants and their training and experience
14 Inspection / cahbration / standardization of equipment
15. Standard Operating Procedures
16. Correspondence
* Minimum requirements
H-16
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Di c mber 4, 19S9
PHASE 3 TECHNICAL GUIDANCE
APPENDD( I; GUIDELINES FOR IDENTIFYING ENFORMA11ON CONCERNiNG UNREASONABLE
ADVERSE EFFECtS PURSUANT TO FIFRA SECTION 4(e)(I)(E)
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December 24. 1Y 9
PHASE 3 TECHNICAL GUIDANCE
GUIDELINES FOR IDENTIFYING INFORMATION CONCERNING UNREASONABLE kD’vERSE
EFFECTS PURSUANT TO FEFRA SECTION 4(e)(l)(E)
What the law requires of registrants
Section 6 (a)(2) of the Federal Insecticide, Fungicide. and Rodenticide Act (FIFRA) states If at
any time after the registration of a pesticide the registrant has additional factual information
regarding unreasonable adverse effects on the environment of the pesticide. he shall submit such
information to the Administrator
Section 4(e)(1)(E) of FIFRA requires a registrant seeking reregistration to submit “an identiflcation
of the data that are required to be submitted to the Administrator under Section 6(a)(2) indicating
an adverse effect of the pesticide
Section 4(e)(4) of FIFRA requires the Administrator to issue guidelines to be follov.ed b
registrants in identifying adverse information Compliance with these guidelines will saiisfy a
registrant’s obligations to submit data pursuant to section 4(e)(1)(E)
2 Section 4(e (1’)(E) Submission
Who Must Submit Information Each registrant who submitted a Phase II response indicating his
intent to reregister his product(s) and who agreed that he is responsible for generating data is
required to dentify and submit adverse effects information required by this guidance document
This incluiks only registrants who submitted Parts A and B in their Phase II response
Deli nit on
“Pesticide uie.ins all active ingredients. inert ingredients, impurities, metaholites. and degrad tes of
the pesticide product for which reregistration is sought
“Formal Re iew” means Special Review, Rebuttable Presumption Against Registration ‘RPAR),
FIFRA section 6(c) proceeding, or FIFRA section 6(b) cancellation proceeding.
“Established Levels” means tolerances, food additive regulations, action levels, or other limitations
on residues imposed by law, reg ilation or other authority.
“Water Reference Level” means the Maximum Contaminant Level (MCL), if one has been
established by EPA. or if it has not, the most recent draft or final long term Health Advisory Level
for the most sensitive subpopulation, such as children. If EPA has published no such levels, the
water reference level is any amount of pesticide detected.
What information must be identified and submitted
Information must be identified and submitted if the registrant, or any officer, employee, or agent
of the registrant possesses or knows of the information.
Information must be identified and submitted if it meets one or more of paragraphs 2 through 10.
Interim test results, raw test data, and other information from an ongoing or uncompleted study
must be identified and submitted if:
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DcLemher 24. 1989
PI-IASE 3 TECHNICAL GUIDANCE
adverse effects which would be reportable in a completed study have been obser ed. and
the registrant has been advised, or a reasonable person would conclude, that the effects
be attributable to exposure to the substance tested, or
the pesticide has been detected in food, feed, and water in amounts that would require the
information to be reported under paragraph 6, if the study were complete.
Opinion Information Opinion information must be identified and submitted if it consists of
opinion(s) or conclusion(s) expressed by a person:
Who was employed or retained (directly or indirectly) by the regiStraflt 2!
From whom the registrant requested the opinion(s) or conclusion(s) in question;
Who by virtue of his knowledge, skill, experience, training, or education could be considered
to be an expert with regard to the matter on which he rendered the opinion.
Clearly erroneous information . Information need not be submitted if prior to the date the Phase
III submission is due to be filed with EPA.
The registrant discovers that any analysis, conclusion, or opinion was predicated on data
that were erroneously generated, recorded, or transmitted, or on computational errors;
E c ’r author of each such analysis, conclusion, or opinion has acknowledged in ri ing that
the analysis, conclusion, or opinion was improper because of the use of the erroneous data.
and has corrected the onginal analysis, conclusion, or opinion accordingly;
As a result of the correction, the information is no longer required to be reportt .1 under
FIFRA section 6(a)(2).
3 Data that must be submitted: toxicological and ecological studies .
The results of a completed study of the toxicity to any human or other non-target organism of a
pesticide must be submitted if the substance tested had a toxic or adverse effect which has not been
previously reported to EPA in a study which meets the requirements of 40 CFR Part 160, or which
in comparison to such a previously submitted valid study shows a toxic or adverse effectS
- in a different organ or tissue of the test organism; or
- at a lower dosage, or after a shorter exposure penod, or after a shorter latency period; or
- at a higher incidence or frequency; or
- in a different species, strain, sex, or generation of test organism; or
- by a different route or medium of exposure; or
- through a different pharmacokinetic, metabolic, or biological mechanism; or
- of greater severity.
However, a study that demonstrates toxic or adverse effect of a pesticide must be identified and
submitted if the pesticide is or has been the subject of a Formal Review.
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December 4. 1989
PHASE 3 TECHNICAL GLIDA,NCE
4 Data that must be submitted Human epidemioLogical studies and residue rnoriitc ring data
Information which concerns any study (or portion thereof) upon which a reasonable person would
conclude that a positi e correlation or association may exist between exposure to a pesticide and
adverse effects in humans, or residues of these pesticides in human tissue or body fluid, must he
submitted regardless of whether or not the registrant considers any observed correlation or
association to be significant
5 Data that must be submitted’ Efficacy studies
Information which concerns any study of the efficacy of a pesticide must be submitted if
- The information demonstrates that the pesticide may not perform in accordance with any
claim by the registrant regarding uses intended for control of organisms which may pose
a hazard to human health, or
The information shows any deficiency or reduction in claimed efficacy of the pesticide, if
the pesticide is, or in the past has been, the subject of a Formal Review
6 Data that must be submitted Studies of pesticides on food or feed, or in water
Food and Feed ’ Information must be submitted if it is from a study which may show that the
pesticide is present on food or feed at a level in excess of established levels, except thai info-mation
on excess residues resulting solely from studies conducted under authority of FIFRA section 5 need
not be suhrn ‘;cd.
Water ’ Information must be submitted if it is from a study which may show that the pesticide.
- Is present in waters of the United States, (as defined at 40 CFR 122 2, except paragraph
(U) of section 122.2) above the “Water Reference Level”, as defined in these guiiclirles.
- Is present is groundwater above the “Water Reference Level”, as defined in these guidelines
Is present in finished drinking water above the “Water Reference Level”, as defined in these
guidelines.
However, information need not be submitted regarding the presence of the pesticide if it is
registered for use in finished drinking water or surface water and the amounts do not exceed those
amounts reported by a registrant, in his application for registration, as resulting from legal
applications of pesticides to water.
7 New metabolites, degradates, contaminants
Information which shows the existence of any substance which appears to be a metabolite, degradate,
or contaminant of a pesticide must be submitted if:
The substance may occur under conditions of use of the pesticide, and
• The existence of the substance or the association of the substance with the pesticide has
not previously been reported to EPA. and
• The registrant has concluded, or has been advised that the substance is of toxicological or
ecological concern, or -
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PHASE 3 TECHNICAL GUIDANCE
- The registrant has concluded, or has been advised that the substance. or analogous
chemicals, may degrade less than 10% in 30 days, or
- A reasonable person would conclude that the Substance may be of toxicological or ecological
concern based on
• the physical chemical properties of the new substance, and/or
- data regarding analogous chemicals, and/or
• data regarding chemical reactivity of the new substance and analogous substances.
and/or
- data ott the new substance.
8 Data that must be submitted. Toxic or adverse effect incident reports
Single incidents : Information regarding a single toxic or adverse effect incident must be submitted
if
- The registrant has been informed that a person or other non-target organism suffered an
adverse effect; and
• The registrant has been informed that the affected person or other non-target organism may
have been exposed to the pesticide, or to one or more of its ingredients, and
• Either
• The registrant has received sufficient information to investigate the reported incident
and has failed to conduct a reasonable investigation pnor to the date of the Phase
III submission; or
- The registrant has concluded or has been advised that there is a causal relationship
between the adverse effect and exposure to its pesticide.
Except ons Information regarding a single toxic or adverse effect need not be submiited if
- It concerns minor irritation of human skin or the eye when the label provides adequate
notice of such a hazard; or
- It concerns non-lethal adverse effects to non-target plants which were at the use s tt at the
time the pesticide was applied.
• It concerns adverse effects to non-target plants which the registrant can conclusively
demonstrate were due to misuse of the pesticide.
Series of incidents . Information concerning any series or pattern of similar incidents attributable
to one of the registrant’s products must be reported unless the causal relationship between the
adverse effect and exposure to the registrant’s pesticide has been conclusively disproved. For
purposes of this section a series or pattern of incidents means 3 or more similar occurrences within
the 5 years preceding submission of information under these guidelines, except, for incidents of
adverse effects to humans, a series or pattern of incidents means 3 or more similar occurrences
within the 10 years immediately preceding submission under these guidelines. Notwithstanding the
above, information pertaining solely to non-lethal adverse effects to non-target plants at the use-
site at the time the pesticide was applied need not be submitted if adequate warnings of the effect
appear on the label of the registrant’s product.
9 Failure of performance incident reports.
Immediate hazard to human health . Information must be submitted which concerns an incident in
which:
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PHASE 3 TECHNICAL GULDA 4CE
• The registrant has been informed that a pesticide product did not pertorm as claimed
against target organisms. and
• The registrant has received sufficient information to conduct an investigation of the incident
and prior to the date of the Phase Ill submission, has not disproved the tailure of
performance, and
- The failure of the pesticide to perform as claimed involved the use against organisms which,
unless controlled, may pose an immediate hazard to human health
Risk to human health . Information must be submitted which concerns a series or pattern of 3 or
more similar incidents in which.
- The registrant has been informed that a pesticide product did riot perform as
claimed against target organisms, and
- The registrant has received sufficient information to conduct an investigation of 3
or more incidents prior to the date of the Phase III submission, and has not proved
that the failures of performance occurred less than 3 times; and
- The failures of the pesticide to perform as claimed involved the use against
organisms which, unless controlled, may pose a risk to human health, and
- The three or more incidents occurred within the 10 years preceding submission p1
information under these guidelines.
Products subiect to Formal Review Information must be submitted which concerns a series or
pattern of three or more individual incidents as to which:
- The rLgistrant has been informed of three of more similar type of failures to pt rf rm as
claimed against target organisms, and
- For at least three individual incidents the registrant has received sufficierii iritormat’on to
conduct an investigation and. prior to the date of the Phase III submission, has tailed to
establish that the reported failure of performance did not occur; and
- The failure of the pesticide to perform as claimed involved any use of a pesticide, if the
pesticide is, or in the past has been the subject of a Formal Review, and
• The three or more incidents occurred within the 10 years preceding submission of
information under these guidelines.
10 Incidents of pesticides on food or feed, or in water
Food and feed . Information must be submitted if it relates to an incident which shows that the
pesticide is present on food or feed at a level in excess of established levels.
Water . Information must be submitted if it is from an incident which shows that the pesticide:
- Is present in waters of the United States (as defined at 40 CFR 122 2, except paragraph
(d) of section 122.2) above the Water Reference Level, as defined in these guidelines.
- Is present in groundwater above the “Water Reference Level”, as defined in these guidelines.
- Is present in finished drinking water above the ‘Water Reference Level”, as defined in these
guidelines.
However, information need not be submitted regarding the presence of the pesticide if it is
registered for use in finished drinking water or surface water and the amounts do not exceed those
amounts reported by a registrant, in his application for registration, as resulting from legal
applications of pesticides to water.
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PHASE 3 TECHNICAL GUIDANCE
ii Reporting of other information
If the registrant knows of information regarding unreasonable adverse effects on the en ironment
of the pesticide, he shall submit such information to the Administrator
12 Note
EPA reserves the right to request additional information regarding exposure or other information
during reregistration phase IV Although EPA will use the above standards as thresholds for
adverse effects under FIFRA section 4(e)(1), during phase IV such information as exceedances of
State Water Quality Standards, monitonng data, or other data in registrants’ possession may be
required.
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