United States
Environmental Protection
Agency
Prevention, Pesticides
and Toxic Substances
(7101)
EPA 712-C-96-348
April 1996
&EPA Ecological Effects Test
Guidelines
OPPTS 850.7100
Data Reporting for
Environmental
Chemistry Methods
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Introduction
This guideline is one of a series of test guidelines that have been
developed by the Office of Prevention, Pesticides and Toxic Substances,
United States Environmental Protection Agency for use in the testing of
pesticides and toxic substances, and the development of test data that must
be submitted to the Agency for review under Federal regulations.
The Office of Prevention, Pesticides and Toxic Substances (OPPTS)
has developed this guideline through a process of harmonization that
blended the testing guidance and requirements that existed in the Office
of Pollution Prevention and Toxics (OPPT) and appeared in Title 40,
Chapter I, Subchapter R of the Code of Federal Regulations (CFR), the
Office of Pesticide Programs (OPP) which appeared in publications of the
National Technical Information Service (NTIS) and the guidelines pub-
lished by the Organization for Economic Cooperation and Development
(OECD).
The purpose of harmonizing these guidelines into a single set of
OPPTS guidelines is to minimize variations among the testing procedures
that must be performed to meet the data requirements of the U. S. Environ-
mental Protection Agency under the Toxic Substances Control Act (15
U.S.C. 2601) and the Federal Insecticide, Fungicide and Rodenticide Act
(7 U.S.C. 136, etseq.).
Public Draft Access Information: This draft guideline is part of a
series of related harmonized guidelines that need to be considered as a
unit. For copies: These guidelines are available electronically from the
EPA Public Access Gopher (gopher.epa.gov) under the heading "Environ-
mental Test Methods and Guidelines" or in paper by contacting the OPP
Public Docket at (703) 305-5805 or by e-mail:
guidelines@epamail.epa.gov.
To Submit Comments: Interested persons are invited to submit com-
ments. By mail: Public Docket and Freedom of Information Section, Office
of Pesticide Programs, Field Operations Division (7506C), Environmental
Protection Agency, 401 M St. SW., Washington, DC 20460. In person:
bring to: Rm. 1132, Crystal Mall #2, 1921 Jefferson Davis Highway, Ar-
lington, VA. Comments may also be submitted electronically by sending
electronic mail (e-mail) to: guidelines@epamail.epa.gov.
Final Guideline Release: This guideline is available from the U.S.
Government Printing Office, Washington, DC 20402 on The Federal Bul-
letin Board. By modem dial 202-512-1387, telnet and ftp:
fedbbs.access.gpo.gov (IP 162.140.64.19), or call 202-512-0135 for disks
or paper copies. This guideline is also available electronically in ASCII
and PDF (portable document format) from the EPA Public Access Gopher
(gopher.epa.gov) under the heading "Environmental Test Methods and
Guidelines."
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OPPTS 850.7100 Data reporting for environmental chemistry meth-
ods.
(a)	Scope—(1) Applicability. This guideline is intended to meet test-
ing requirements of the Federal Insecticide, Fungicide, and Rodenticide
Act (FIFRA) (7 U.S.C. 136, et seq.) and the Toxic Substances Control
Act (TSCA) (15 U.S.C. 2601).
(2) [Reserved]
(b)	Data reporting. (1) Registrants must provide EPA with accurate
and precise data on the performance of those soil and water chemistry
methods that are used to develop laboratory and/or field residue data to
support exposure, environmental fate, and ecological effects studies for
registration and reregistration. This includes all single or multianalyte
methods for the parent compounds, toxicologically significant metabolites,
and degradates in each environmental matrix. Registrants shall provide
EPA with clearly written and complete analytical methods that describe
the exact procedure, materials, and equipment to be used by regulatory
chemists to validate their methods. Analytical methods shall be practical,
rapid, and quantitate analytes of interest. The Agency will accept best
available methods for those parent compounds, toxicologically significant
metabolites, and degradates that have significant risks and require state-
of-the-art equipment to measure trace amounts of analytes. Both practical
and rapid and best available methods must use equipment that is commer-
cially available in the United States. If methods use conventional gas chro-
matography, liquid chromatography, etc., the registrants shall submit con-
firmatory methods using GC/MS, LC/MS, second column or other suitable
procedures. GC/MS and LC/MS methods that are used as the primary
method to generate study data do not require another confirmatory proce-
dure. Regulatory chemists should be able to validate practical and rapid
analytical methods using a set of samples in twenty-four hours (e.g. three
eight-hour working days); however, EPA recognizes that best available
methods may require additional time.
(2) EPA regulatory chemists should be able to quickly evaluate soil
and water methods for each test level and matrix. Registrants shall submit
performance data to EPA demonstrating that an adequate number of sam-
ples for each test level were extracted, cleaned up, and analyzed. Each
set should have an appropriate number of samples with quality control
samples intermingled. This data should support the limit of detection
(LOD) or the method detection limit (MDL), the established limit of quan-
titation (LOQs) and precision and accuracy for each method. The mean
recovery at each spiking level, at or above the LOQ, should lie between
70 and 120 percent of the known quantity of the pesticide/metabolite/
degradate spiked into the matrix blanks during the method validation. Reg-
istrants are to report individual values for recoveries, standard deviations
and confidence limits for the pesticide parent, significant metabolites and
degradates in fortified samples at each spiking level. The relative standard
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deviation (RSD) of replicate measurements of recoveries should not exceed
the target level of plus or minus 20 percent at or above the LOQ for each
spiking level. EPA recognizes that some methods may not be able to meet
these precision requirements. The registrant shall describe how they cal-
culated the values for MDLs or LODs and LOQs and cite any references
that they believe are appropriate. Registrants shall not correct sample val-
ues for recoveries. Registrants shall also describe any matrix or solvent
effects that result in signal enhancement, masking or suppression and the
impact those effects have on the test results. Registrants shall clearly iden-
tify those laboratories that developed the data for the soil and water meth-
ods they submit to EPA. Registrants shall use an independent lab to vali-
date (ILV) soil and water methods that are used to support appropriate
exposure, environmental fate and ecological effects studies. A separate re-
port will be prepared by the registrant for the ILV testing and submitted
to EPA with the appropriate study. Methods will not be rejected outright
for failure to comply with each and every aspect of this DRG but will
be reviewed on a case-by-case basis to determine their suitability by
science reviewers and chemists in OPP.
(c) Data reporting format. The following suggested reporting format
is presented to aid the applicant/registrant in generating reports which are
compatible with the Agency's review process.
(1)	Title/cover page.
(2)	Certification, (i) Certification of authenticity by the Study Director
(including signature, typed name, title, affiliation, telephone number, and
date, etc.).
(ii)	Statement of adherence to the FIFRA GLP.
(iii)	Statement of claims for non-confidentiality and that the method
contains no trade secrets or proprietary data.
(3)	Table of contents.
(4)	Summary. Provide a brief description of paragraphs (c)(6)(ii) and
(c)(6)(iii) of this guideline.
(5)	Materials, (i) Equipment (list and describe).
(ii)	Reagents and standards (provide MSDS sheets and list and de-
scribe the source, purity and stability of analytical grade standards; de-
scribe the source and preparation of reagents.
(iii)	Safety and Health (describe any special precautions that need
to be taken with solvents or reagents and any procedural steps that require
special precautions to avoid safety or health hazards).
(6)	Methods, (i) Principles of the analytical methods.
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(ii)	Analytical procedures (describe in a detailed step wise fashion):
(A)	Source and characterization of soil samples (e.g. soil (textural
class and percent organic matter)) and water samples (pH, turbidity) etc.
(B)	Preparation of samples.
(C)	Extraction (demonstrate efficiency in specific soil samples from
field sites).
(D)	Fortifications (i.e. during method validation runs).
(E)	Clean-up.
(F)	Derivatization (if any).
(iii)	Instrumentation. (A) Description (e.g. make/model, type/specifity
of detector(s), column(s) (packing materials, size), carrier gas(es), etc.
(B)	Operating conditions (e.g. flow rate(s), temperature(s) voltage,
etc.).
(C)	Calibration procedures (frequency and stability).
(iv)	Potential interference(s) (describe the effects of the following on
signal enhancement, masking or suppression of signal and their impact
on the test results):
(A)	Sample matrices.
(B)	Other pesticides used on the test site.
(C)	Solvents.
(D)	Labware.
(v)	Confirmatory techniques. Describe confirmatory techniques, i.e.
GC/MS, LC/MS, second column, etc.
(vi)	Time required for analysis. Give the time required to take a sam-
ple/set completely through the analytical procedure, including sample prep-
aration, extraction, cleanup, derivatization, and determination steps. Iden-
tify those steps that could significantly increase the time required to com-
plete the method.
(vii)	Modifications or potential problems. Specify any allowable
variances and describe any unique steps where little variation is allowed
in the method. Describe any potential problems and/or modifications that
were made to the analytical procedures.
(viii)	Methods of calculation. Describe calculations in a step wise
fashion and include calibration factors, calibration curves for parent
compound, metabolites, and degradates, etc.
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(ix)	Copies of chromatograms/spectra. Representative sample
chromatograms/spectra should be submitted for each analyte measured in
each matrix at all spiking levels, including method and matrix blanks. In
addition, chromatograms/spectra with the highest levels of background and
poorest resolution will also be sent to EPA. Copies of the chromatograms/
spectra are also required for the standards that were used to quantitate
the analyte(s) in the representative matrix chromatograms/spectra submit-
ted to EPA.
(x)	Other. Describe any and all additional information the registrant
considers appropriate and relevant to provide a complete and thorough de-
scription of the soil and water method.
(7)	Results/discussion, (i) Method validation results (include tables
of test levels and results of analysis).
(ii)	Accuracy (mean, range of recoveries, standard deviations and con-
fidence limits for specific concentration levels, such as the LOQ or 10X
the LOQ).
(iii))	Precision (relative standard deviation at specific concentration
levels).
(iv)	Limit of detection (provide definitions).
(v)	Limit of Quantitation (provide definitions).
(vi)	Ruggedness testing (if performed).
(vii)	Discussion of selectivity and specificity of method.
(viii)	Limitations.
(ix)	Independent laboratory validation.
(8)	Conclusion. Discuss applicability of the soil and water method
for measuring specific test compound(s) in various matrix(ces), ranges, ex-
pected recoveries, interference(s), etc.
(9)	Tables/figures.
(10)	References.
(d) Independent laboratory validation—(1) Guidelines for the reg-
istrant. (i) The registrant should apply the same criteria of quality in se-
lecting an independent laboratory (IL) for environmental chemistry method
validations as they would for any other analytical work. An independent
lab can be privately or publicly owned or in the registrant's own organiza-
tion. If that lab is located in the registrant's organization, or is in anyway
associated with the development of the original environmental chemistry
method (ECM), the same people, equipment, instruments, and supplies (i.e.
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glassware, solvents, reagents, standard reference materials) should not be
utilized to validate the ECM. The Agency does not expect the registrant
to synthesize or purchase another lot of authentic grade analytical standard
but it does expect the independent lab to use a new aliquot of that standard
in order to prepare spiking solutions for the ILV.
(ii)	Furthermore, the personnel conducting the ECM validation should
not report to the same study director who was involved in developing the
original method or who may have used the method to develop data for
field studies to support pesticide registration or reregistration actions. The
lab chemists chosen to conduct the independent laboratory validation must
be unfamiliar with the method both in its development and subsequent
use in analyzing samples collected from field studies. Those individuals,
however, should be trained and experienced pesticide residue chemists.
The ECM given to the independent lab should be the same one that was
used by the registrant to generate field study data. Any significant changes
that are made to that method by the chemists at the independent lab should
be incorporated into the original method and reported in writing imme-
diately to the registrant. If those changes impact the performance of the
original method, such as its precision, accuracy or limit of quantitation,
the registrant should report those changes to the Agency.
(iii)	Soil and water samples should be identified and/or supplied to
the independent lab by the registrant; however, chemists at the independent
lab should use those samples to prepare matrix spikes for the validation
study. Registrants should select the most difficult matrix to analyze from
the appropriate field study identified on the front of this document to dem-
onstrate how the method performs.
(2) Guidelines for the independent laboratory, (i) The laboratory
conducting the ILV must use the method exactly as it is written by the
registrant or another private or public laboratory and should only contact
the registrant or developers to clarify minor deficiencies in the method.
For example, if the characteristics of the clean up column are not ade-
quately described in the method, the independent laboratory should contact
the registrant or developers for clarification. EPA recognizes that chemists
in the independent laboratory need to establish the method and that they
will need to compute the instrument detection limits and determine the
retention times of the analytes.
(ii) They will also need to establish the relationship between the in-
strument responses and concentrations of analytes and to verify that matrix
control samples are free of interferences at the appropriate retention time,
wavelength or detector setting. All quality control conditions must be satis-
fied in order to demonstrate that the method is under control before the
IL analyzes any performance samples to be reported to EPA. Any contact
with the registrant or developers during the establishment of the method
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must be documented in writing in the final report submitted by the inde-
pendent laboratory.
(iii)	The independent laboratory (IL) should be able to quickly evalu-
ate soil and water methods for each test level and matrix. ILs shall submit
performance data to registrants demonstrating that an adequate number of
samples for each test level were extracted, cleaned up, and analyzed. Each
set should have an appropriate number of spiked matrix samples with a
method and matrix blank intermingled. This data, and subsequent computa-
tions, should support the registrant's limit of detection (LOD) or the meth-
od detection limit (MDL), the established limit of quantitation (LOQs) and
precision and accuracy for each method. Individual recoveries at each spik-
ing level, at or above the LOQ, should lie between 70 and 120 percent
of the known quantity of the pesticide, metabolite and degradate spiked
into the matrix blanks during the method validation. Independent labs are
to report mean and individual values for recoveries and standard deviations
for the pesticide parent, toxicologically significant metabolites and/or
degradates in fortified samples at each spiking level. They should also
report the confidence intervals (95 or 99%) for the true average recoveries
at each spiking level. The relative standard deviation (RSD) of replicate
measurements of pesticide concentrations should not exceed the target
level of less than or equal to 20 percent for each spiking level at or above
the LOQ. EPA recognizes that some methods may not be able to meet
these precision requirements. The IL will use the predetermined values
from the registrant for LOQs and 10XLOQ to establish appropriate spiking
levels. ILs shall not use data from matrix controls (blanks) to correct val-
ues from spiked matrix blanks for recoveries. Interferences with peak areas
that are less than fifty percent at the MDL/LOD, are considered negligible.
ILs shall also describe any matrix or solvent effects that result in signal
enhancement, masking or suppression and the impact those effects have
on the test results. ILs will prepare a well documented laboratory report
to send to the registrant for the ILV and that report with appropriate
changes recommended by the registrant will be submitted to EPA with
the field study.
(iv)	Independent labs will be allowed to analyze three sample sets
in order to validate the method as written. A complete set should consist
of a reagent blank, two unspiked matrix control samples and five matrix
control samples spiked at the LOQ and another five matrix control samples
spiked at 10XLOQ for each distinct matrix. A complete set may include
more than thirteen samples depending on the number of reagent, unspiked
and spiked control matrix samples. It may be necessary, however, to divide
a complete set into two subsets for efficient handling. Each subset should
contain a reagent blank, two unspiked matrix control samples and five
matrix control samples spiked at the LOQ or 10XLOQ.
(v)	If the performance data on the first set of samples at any of the
required spiking levels is unsuccessful, the independent laboratory may
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contact the registrant to clarify the directions given in the method. Any
contact with the registrant or developers during the method validation must
be documented in writing in the final report submitted by the independent
laboratory. If the independent laboratory cannot generate performance data
that is similar to the registrant's or developers' after the second set of
spiked samples, the independent laboratory may contact the registrant to
further clarify the directions given in the method. If the independent lab-
oratory cannot generate performance data that is similar to the registrant's
or developers' after the third set, the method should be failed and a report
will be sent to the registrant explaining why the method failed. The reg-
istrant should then decide whether to repeat the independent laboratory
validation at another laboratory, further develop the method or withdraw
it.
(vi) A successful ILV will require performance data on at least one
complete set of samples that meets those criteria described in paragraphs
(d)(2)(iii) and (d)(2)(iv) of this guideline. The revised or rewritten method,
performance data, chromatograms and computations from all sets and/or
subsets will be sent to the registrant. The registrant, with the assistance
of the IL, will decide how to present that information to EPA.
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