United States Prevention, Pesticides EPA712-C-96-174
Environmental Protection and Toxic Substances August 1996
Agency (7101)
&EPA Residue Chemistry
Test Guidelines
OPPTS 860.1340
Residue Analytical
Method
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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
(7U.S.C. I36,etseq.).
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), internet: http://
fedbbs.access.gpo.gov, or call 202-512-0132 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 860.1340 Residue analytical method.
(a) Scope—(1) Applicability. This guideline is intended to meet test-
ing requirements of both the Federal Insecticide, Fungicide, and
Rodenticide Act (FIFRA) (7 U.S.C. 136, et seq.) and the Federal Food,
Drug, and Cosmetic Act (FFDCA) (21 U.S.C. 301, et seq.).
(2) Background. The source material used in developing this har-
monized OPPTS test guideline is OPP 171-4 Results of Tests on the
Amount of Residue Remaining, Including A Description of the Analytical
Methods Used (Pesticide Assessment Guidelines, Subdivision O: Residue
Chemistry, EPA Report 540/9-82-023, October 1982). This guideline
should be used in conjunction with OPPTS 860.1000, Background.
(b) Purpose. (1) Based on plant and animal metabolism study results,
the Agency requires tolerance petitioners to develop analytical methods
to determine all components of the total toxic residue (TTR). In some
cases, it is not possible to develop a single method that can determine
all components of the residue, and several methods are required. Residue
analytical methods are used to obtain residue data on which dietary expo-
sure assessments and tolerances are based, and to enforce the tolerance
after it is established. Enforcement methods are validated by an independ-
ent laboratory before submission to the Agency as required by PR Notice
96-1 (see paragraphs (c)(6) and (e)(3) of this guideline). The Agency vali-
dates each new analytical method using a method trial to ensure that the
procedures are appropriate for tolerance enforcement.
(2) The methods for residue analyses should serve two functions:
They must provide the residue data upon which judgements are made as
to the identity and magnitude of residues resulting from the proposed use,
and they must provide a means for enforcement of the tolerance. The meth-
ods described in the FDA Pesticide Analytical Manual (PAM), Volume
II, and the Official Methods of Analysis of the Association of Official
Analytical Chemists (as referenced in paragraphs (e)(7) and (e)(l) of this
guideline) can be used as examples of suitable analytical methods.
(c) Test method—(1) General, (i) The analytical methods must be
described in a stepwise fashion in sufficient detail to enable competent
analysts to apply the method even though they are unfamiliar with the
procedure. Residue analytical methods should be practical, rapid, and
quantitate the TTR in the tolerance expression. While the Agency, on a
case-by-case basis, may accept best-available methods for the TTR that
require state-of-the-art equipment, the equipment must be commercially
available in the United States. Reprints of published methods may be sub-
mitted. However, where modifications have been made to adapt a basic
method to other crops for which a tolerance is proposed, details of the
modifications are needed. This includes application to processed byprod-
ucts and meat, milk, poultry, or eggs, if these are a consideration.
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(ii) The method should not be subject to substrate-related inter-
ferences or those arising from reagents. Appropriate clean-up measures
should be incorporated to reduce or eliminate spurious responses that
might jeopardize the results. For example, in gas-liquid chromatographic
(GLC) methods, separation should be sufficiently distinct to yield reason-
ably discrete peaks for the components of interest rather than a response
which appears as a shoulder on an interfering peak.
(iii) The Agency encourages submission of more direct and easily
performed methods for tolerance enforcement. However, the subject meth-
ods must meet the Agency's stringent criteria for enforcement procedures
(see paragraph (c)(5) of this guideline). Although methods used solely for
data collection do not need to meet all the requirements of enforcement
methods, they must be validated in a similar manner to assure the Agency
they are adequate for measuring the TTR.
(iv) The use of the FDA multiresidue methods (MRMs) in PAM, Vol-
ume I, under paragraph (e)(6) of this guideline, as primary enforcement
methods is encouraged. Petitioners are required to submit MRM test data
for the parent compound and all regulated metabolites (see OPPTS
860.1360). It is recommended that petitioners determine the suitability of
the MRMs for measuring a new pesticide prior to developing a single-
analyte method. If one of the MRMs is found to be acceptable as the
enforcement method, an independent laboratory validation as described in
paragraph (c)(6) of this guideline will not be required. However, the peti-
tioner will still be expected to provide a single analyte confirmatory meth-
od to be published in PAM, Volume II (see paragraph (e)(7) of this guide-
line).
(v) Whenever possible, GLC retention times and response values
should be reported relative to those of a stable reference compound, par-
ticularly when the residue is converted to a derivative prior to gas chroma-
tography. GLC and high-performance liquid chromatography (HPLC) pa-
rameters should be reported in a form such as the guidelines outlined for
the multiresidue method (MRM) protocols found under paragraph (e)(6)
of this guideline.
(2) Validation of method by petitioner, (i) Methods must be vali-
dated by control sample data and recovery data for all components of the
TTR on an adequate representation of the commodities involved. Control
values should be reasonably low in relation to the proposed tolerance, pref-
erably less than 20 percent of the proposed tolerance. Recoveries should
be at fortification levels appropriate to the proposed tolerance and should
include the limit of quantitation (LOQ). Recoveries should lie between
70 percent and 120 percent of the known quantity of the pesticide and
its metabolites spiked into the substrate controls, and should not vary sig-
nificantly from sample to sample. Methods in which recoveries are consist-
ently greater than 100 percent are considered questionable. If 70 percent
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recovery is not attainable, the Agency will accept, on a case-by-case basis,
methods having lower recoveries for active ingredients that are not acutely
toxic or for minor metabolites.
(ii) Petitioners are to report individual values for recoveries, standard
deviations, and confidence limits for all components of the TTR. The resi-
due levels being measured is a major factor in determining the acceptable
variability of a method. Appropriate coefficients of variation (CVs) or rel-
ative standard deviations (RSDs) as a function of residue level are dis-
cussed in references such as paragraph (e)(5) of this guideline. The Agency
will consider the variability in recovery values when determining the ac-
ceptability of methods with recoveries outside the 70-120 percent range.
For example, a method with average recovery of 65 percent and a low
CV (e.g., 5 percent) may be viewed more favorably than a method show-
ing 95 percent average recovery and a CV greater than 20 percent.
(iii) The raw agricultural commodity (RAC), or a macerate thereof,
should be fortified, rather than crop, animal tissue, milk, or egg extracts.
The portion of the crop to be analyzed is specified under paragraph (e)(6)
of this guideline, in 40 CFR 180.1, and in Table 1 in OPPTS 860.1000.
Petitioners are also advised to consult the proposed regulation (40 CFR
180.45) on portion of food commodities to be analyzed (see (e)(6) of this
guideline). The petitioner should state the estimates of the practical limits
of detection (LOD) and quantitation as applied to each of the subject crops
or tissues. The estimates of the practical LOD and LOQ should be based
on the least concentration of pesticide which can be detected or quantitated
with a reasonable degree of assurance, taking into account the size and
variation of blanks (instrument response due to crop extractives and re-
agents). The petitioner should describe how the values for LOD and LOQ
were calculated and cite any appropriate references.
(iv) 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 (e.g. root and tuber vegetables, leafy vegetables (except
Brassica vegetables), and cereal grains), the method needs to be validated
on only the representative crops for the group as specified in 40 CFR
180.41. The report submitted on the method itself needs to include recov-
ery data on only a representative number of crops. However, in crop field
trial reports, additional validation data should be provided on any crop
that was not tested for the analytical method report.
(v) With respect to animal commodities, validation data are required
for milk, eggs, and all tissues for which residue data are collected in feed-
ing studies and/or for which tolerances need to be established. The tissues
normally include cattle muscle, fat, liver, and kidney and poultry muscle,
fat, and liver. The recovery data for cattle commodities will in most cases
cover the products of goats, hogs, horses, and sheep.
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(3) Extraction efficiency, (i) Conventional recovery experiments as
discussed under paragraphs (c)(2) and (c)(3) of this guideline do not nec-
essarily reflect the efficiency with which aged (weathered) residues are
extracted from crops. There should be some assurance that aged residues
are completely extracted by the procedure. Utilization of radiolabeled sam-
ples from the plant and/or livestock metabolism studies (referred to as
radiovalidation of the method) provides the best evidence on completeness
of extraction. Analytical methods should be radiovalidated to determine
whether the TTR is extracted from weathered plant matrices and/or live-
stock tissues, milk, or eggs. Samples should undergo the extraction proce-
dure employed in the method. The petitioner needs to demonstrate that
the extracted radioactivity accounts for most of the TTR that was identified
in the metabolism study. This may involve performing the determinative
step of the method or other procedures which will identify and quantitate
the components of the extracted radioactivity. Simply quantitating the total
extracted radioactivity will not suffice in most cases. If a method is to
be used on both plant and animal commodities, it should be radiovalidated
on a plant matrix, an animal tissue, and either milk or eggs. Matrices for
which extraction is expected to be most difficult should be used. In the
case of plants this would normally be a dry sample (e.g. straw, fodder)
containing residues which have been on the plant for a relatively long
period. Petitioners should provide a rationale for the samples used in the
radiovalidation. If the data collection and enforcement methods have sig-
nificantly different extraction steps, each method should be radiovalidated.
Alternatively, analyses of several split samples bearing field weathered,
nonradiolabeled residues showing similar results with the two methods
may be submitted.
(ii) Methods utilizing only surface stripping are not acceptable except
for commodities where extensive data have established that the TTRs are
in fact only surface residues.
(iii) Certain components of the TTR may occur bound with naturally
occurring plant constituents, and thus may not be recovered by extraction
techniques that are satisfactory for the free components. Whenever there
are indications of the formation of bound components which may not be
recovered by the extracting solvent but are of toxicological concern, modi-
fications should be made in the procedure that will free and recover the
liberated components. One such modification would be the initial hydroly-
sis of the treated crop under acidic, basic, or enzymatic conditions to free
the components. In some cases these bound components may also be re-
covered with polar solvents. These bound residues should not be confused
with those fragmentary components which may be so tightly bound or
incorporated into the plant's metabolic pool that they are not recoverable
by any chemical means. Such components are of interest, but are not usu-
ally of toxicological concern. Refer also to the discussion on nonextract-
able residues in OPPTS 860.1300.
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(4) Determination of TTRs. (i) The methods employed should meas-
ure the TTR found in the metabolism studies outlined in OPPTS 860.1300,
Nature of the Residue. Often all components of toxicological concern will
contain a common chemical moiety so that the method may be adapted
to determine all compounds simultaneously. However, in some cases, it
may be necessary to adapt separate extraction-cleanup procedures, or even
another complete method, to measure the TTR or a significant component
of the residue. In other cases, one or more components of the residue
will be significantly more toxic than other components of the residue and
will have to be determined separately.
(ii) In some cases the Agency will accept a tolerance enforcement
method that measures only a portion (typically the parent compound) of
the TTR in order to ease burdens on enforcement agencies and/or to har-
monize with international maximum residue limits. This may be referred
to as an indicator or marker compound. However, in order to have suffi-
cient data for dietary risk assessment, a data collection method will nor-
mally still be needed to quantitate the TTR. Petitioners contemplating use
of an indicator or marker compound in either enforcement or data collec-
tion methods are advised to contact the Agency about the acceptability
of this approach.
(5) Requirements for regulatory methods, (i) One or more of the
methods proposed in the petition must be acceptable to enforce the pro-
posed tolerance. Where applicable, use of the FDA multidetection meth-
odology outlined in paragraph (e)(6) of this guideline is strongly rec-
ommended. Also, the enforcement method should be as simple as possible
to decrease the cost of monitoring for pesticide residues.
(ii) A method which may be valid for gathering residue data is not
necessarily suitable for enforcement purposes. In general:
(A) An enforcement method should not require:
(7) The use of a sample of the untreated commodity as a blank.
(2) Exotic equipment or reagents (or reagents that are no longer man-
ufactured).
(B) An enforcement method should be:
(7) Reasonably rapid in execution. In general, residue analytical meth-
ods for regulatory purposes should require one working day for comple-
tion. Methods taking longer than one working day will be considered ac-
ceptable on a case-by-case basis. Methods taking less than 24 hours (total
time from initiation to completion of analysis) will be required for acutely
toxic residues because of the possibility of enforcement action from acci-
dent or misuse situations.
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(2) Sufficiently specific to measure and identify the residue in the
presence of residues of other pesticides which could reasonably be ex-
pected to be present on the same commodity.
(3) Sufficiently sensitive in relation to the tolerance proposed.
(4) Practicable without the use of extremely hazardous or toxic re-
agents.
(iii) The Agency does not have a specific list of the types of analytical
methods which are acceptable for enforcement of tolerances. Any proce-
dure (e.g. GC, HPLC, MS, immunochemical) which meets the criteria de-
scribed above would be acceptable. Methods based on cholinesterase inhi-
bition are not regarded as suitable for enforcement purposes. Although
methods based on paper or thin layer chromatography which visually
measure the residue may not be adequately quantitative for enforcement
purposes, they may be useful as confirmatory methods to help identify
the residue.
(iv) Although certain gas and liquid chromatographic detection sys-
tems possess inherent specificity, methods based on these systems should
usually be supplemented by a confirmatory method which is significantly
different from the primary enforcement method. In general, confirmation
by mass spectrometry is suitable. The specificity may also be enhanced
by the use of special extraction-cleanup procedures, derivatization, or par-
allel and/or alternate columns. Provided that a specific confirmatory meth-
od is available, the Agency will not require that an interference study be
conducted to show whether other pesticides registered on the same com-
modities interfere with determination of the pesticide of interest.
(v) The Agency accepts use of common moiety methods on a case-
by-case basis. Toxicological differences among all metabolites of concern
which can be determined by the method are taken into consideration when
evaluating the suitability of a common moiety method. In those cases
where a common moiety method is proposed as the primary enforcement
method and other regulated pesticides produce the same common moiety,
a confirmatory method specific for the residue of concern should be avail-
able to enforcement laboratories. This is especially critical in those in-
stances where two pesticides generating a common moiety are registered
on the same crop but have different tolerance levels.
(vi) The methods proposed for enforcement may be subjected to trials
in Agency laboratories if the pesticide is new, if the analytical methods
is new and unfamiliar, or if the commodity is known to be difficult to
analyze. The burden of proof is on the petitioner, and should the method
fail to perform as expected in these trials, the petitioner will be asked
to resolve the difficulties. Also, the petitioner will be responsible for im-
proving such a method and furnishing new residue data by the improved
method. If the method performs satisfactorily and is acceptable as an en-
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forcement method, it will be made available to interested parties by publi-
cation or reference in the PAM under paragraph (e)(7) of this guideline.
Thus, a petition must include a copy of the analytical method which is
not claimed to be, or stamped, confidential business information. Methods
will be released to enforcement agencies prior to the establishment of a
permanent tolerance if a FIFRA section 18 emergency exemption or a tem-
porary tolerance is in effect. Prior to publication in the PAM under para-
graph (e)(7) of this guideline, methods can be obtained directly from the
Agency.
(6) Independent laboratory method validations. This paragraph de-
scribes performance of independent laboratory validation (ILV) trials for
submission as part of the pesticide petition.
(i) ILV trials required to accompany petitions for tolerance. Results
of ILV trials of new analytical methods are required for the parent pes-
ticide, including metabolites of toxicological concern, and must accompany
the following types of petitions:
(A) The first tolerance petition including temporary tolerance petitions
for residues of a pesticide in a RAC or processed food/feed.
(B) Any new tolerance petition for residues of a pesticide with pre-
viously established tolerances if a new method is proposed for enforce-
ment.
(C) Any new tolerance petition for residues of a pesticide with pre-
viously established tolerances if the previously approved enforcement
method has been significantly modified to accommodate the new commod-
ity. If the petitioner is uncertain whether a method change is significant,
the Agency should be contacted.
(ii) Results of an ILV trial are usually not required for an enforcement
method which the Agency deems superior to the currently accepted en-
forcement method. An ILV trial is also not normally required for confirm-
atory methods. However, at the discretion of the Agency, an ILV trial
may be required for confirmatory methods on a case-by-case basis. One
particular instance when the ILV trial is likely to be needed is for a con-
firmatory method associated with a compound whose primary enforcement
method is a common moiety procedure which also detects other registered
pesticides.
(iii) The laboratory personnel including the study director chosen to
conduct the ILV trials must be unfamiliar with the method, both in its
development and in its subsequent use in analyzing field samples. Provided
this criterion is met and the same equipment, instruments, and supplies
are not used, the laboratory chosen to conduct the ILV trials may be in
the petitioner's organization. Other possibilities include laboratories at
state enforcement agencies, at universities, or private laboratories. The pe-
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titioner should apply the same criteria of quality in selecting a laboratory
for ILV trials as would be done for any analytical work.
(iv) Requirements for ILV trial. (A) ILV trials must be conducted
under FIFRA Good Laboratory Practice (GLP) standards as specified in
40 CFR part 160. A successful ILV trial will require adequate results for
the TTR on at least one set of samples, and the laboratory conducting
the ILV trial will be allowed to run up to three sets of samples using
the method on a given commodity. A set of samples consists of two con-
trol samples, two control samples fortified at the proposed tolerance, and
two control samples fortified at the LOQ. If the tolerance is proposed at
the LOQ, the second fortification level should be twice the LOQ. At the
discretion of the petitioner, one additional fortification at another level may
be included in the set of samples. The method must be performed as writ-
ten with no significant modifications. If additional commodities are ana-
lyzed by the same method, they will be considered to be separate ILV
trials.
(B) The laboratory conducting the ILV trial may contact the devel-
opers or previous users of the method prior to running the first set of
samples, but all communications must be logged and reported to the Agen-
cy. Under no circumstances should anyone from the petitioner, developer,
or any previous users be allowed to visit the laboratory during the ILV
trial to observe, offer help, or assist the chemists or technicians. If the
first (or second) set is not successful, and the laboratory requires additional
contact with the developers or other users of the method, all communica-
tions should be recorded. Any subsequent additions or modifications to
the original method resulting in improved performance should be incor-
porated into the method write-up that is sent to the Agency for validation.
(C) If one method is to be used for several commodities, the ILV
trial should be carried out on that commodity the petitioner has had the
most difficulty analyzing. If the same method is used for both plant and
animal commodities, separate ILVs should be run on both the most dif-
ficult plant and most difficult animal matrices. The rationale for selection
of the commodity should be provided. If, after three sets of samples have
been analyzed, the ILV trial has failed to produce adequate results, the
petitioner must revise the method and perform a second confirmatory trial
using a different laboratory.
(D) The results on one set of samples, after conducting no more than
three sets, must be similar to those achieved by the petitioner to constitute
a successful ILV trial. Recovery rates should be 70-120 percent and inter-
ference should be negligible compared to the proposed tolerance level.
(v) Information to be reported to the Agency. If the ILV trial is suc-
cessful, the following should be submitted by the petitioner:
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(A) Name, address, and telephone number of study director and other
contact person for ILV laboratory.
(B) Description of the analytical method.
(C) All recovery and control values for all commodities that were
obtained during all ILV trials.
(D) Representative chromatograms/spectra for each analyte in each
matrix.
(E) Description of the instruments used and operating parameters.
(F) Description of any problems encountered and a written description
of any changes or modifications that were made during the ILV.
(G) Any steps considered critical, i.e. steps where little variation is
allowable or directions must be followed precisely.
(H) The number of worker-hours required to complete one set of sam-
ples.
(I) The number of calendar days required for one set of samples.
(J) Any contact between the independent laboratory and the method
developers or others familiar with the method, including the reasons for
the contact, any changes in the method that resulted, and the time of this
communication with respect to the progress of the confirmatory trial (i.e.
after the first set, during the second set, etc.).
(K) A statement of adherence to FIFRA GLP standards under 40 CFR
160.12.
(vi) The Agency will continue to conduct method validations. If the
Agency determines that the petitioner has submitted results of a successful
ILV trial by an independent laboratory, the method will be validated by
Agency chemists.
(7) Internal and procedural standards, (i) The Agency accepts the
addition of an internal standard to the final extract just prior to injection
to serve as a calibration for retention times and/or peak heights/areas and
to improve the precision of quantitation. 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 that the analyte and internal standard behave identically in each
step (extraction, cleanup, etc.). For chromatographic methods, the peaks
for the analyte and internal standard should elute close to one another,
but be well resolved from each other. An example of an internal standard
method which the Agency has accepted is the use of isotopically labeled
dioxins for dioxin analysis by mass spectrometry (see paragraph (e)(2) of
this guideline). As with any other reagent or reference standard used in
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an enforcement method, the internal standard must be available to enforce-
ment laboratories. If an internal standard is not commercially available,
the petitioner must ensure a supply of the chemical to the Agency.
(ii) Procedural standards are considered to be standards which are
generated by subjecting the reference standard to some or all of the sample
preparation procedures specified in the method. The Agency will accept
methods using procedural standards generated from a derivatization step
under certain conditions. If a procedural standard is used, the petitioner
should supply the Agency with not only the pesticide analytical standard,
but also the derivatized standard. Availability of the derivatized standard
would allow the enforcement laboratory to determine the efficiency of the
standard preparation. If the derivatized standard is unstable or cannot be
provided, the petitioner must provide data to demonstrate the efficiency
and reproducibility of the procedure.
(d) Data reporting format. The following format is suggested for
data reporting. However, other formats are also acceptable provided the
information described in this paragraph is included.
(1) Title/Cover page. Title page and additional documentation require-
ments (such as requirements for data submission and procedures for claims
of confidentiality of data) if relevant to the study report should precede
the content of the study.
(2) Table of contents. The table of contents should provide page num-
bers for the essential elements of the study:
(3) Body of report—(i) Introduction and summary. (A) Scope (suit-
able matrices) and source of method (e.g. the PAM, company reports, etc.).
(B) Description of the principles of the analytical procedure, including
identification of the chemical species determined and the limits of detec-
tion and quantitation.
(ii) Materials and methods. (A) Equipment—list and describe.
(B) Reagents and standards—list and describe source and preparation.
(C) Analytical procedure—detail in a stepwise fashion, with special
emphasis on reagents or procedural steps requiring special precautions to
avoid safety or health hazards.
(7) Preparation of sample.
(2) Extraction—demonstrate efficiency, if relevant (e.g. dry crop sub-
strates, bound residues, etc.); radiovalidation data may be provided in a
separate report at the discretion of the petitioner.
(3) Fortification, if applicable—i.e. during method validation runs.
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(4) Clean-up.
(5) Derivatization (if any).
(D) Instrumentation—include information on:
(7) Description (e.g. make/model, type/specificity of detectors, col-
umns (packing materials, size), carrier gases, etc.).
(2) Operating conditions (e.g., flow rates, temperatures, voltage, etc.).
(3) Calibration procedures.
(E) Interferences—describe tests.
(7) Sample matrices.
(2) Other pesticides.
(3) Solvents.
(4) Labware.
(F) Confirmatory techniques—describe.
(G) Time required for analysis (to carry a sample/set completely
through the analytical procedure, including the determinative step).
(H) Modifications or potential problems, if any, in the analytical
methods (detail circumstances and corrective action to be taken).
(I) Methods of calculation (describe in a stepwise fashion).
(7) Calibration factors.
(2) Analyte in sample.
(J) Other. Any and all additional information the petitioner considers
appropriate and relevant to provide a complete and thorough description
of residue analytical methodology and the means of calculating the residue
results.
(iii) Results and discussion. Describe expected performance of meth-
od.
(A) Accuracy (expected mean and range of recoveries)—include,
preferably in tabular format, the individual recovery values, average recov-
eries, and relative standard deviation thereof for each component of the
TTR in each commodity tested during the petitioner's method validation.
(B) Precision.
(C) Limits of detection and quantitation (provide definition).
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(D) Ruggedness testing, if performed.
(E) Limitations.
(iv) Conclusions. Discuss applicability of analytical procedure for
measuring specific test compounds in various test substrates, ready avail-
ability of equipment, interferences, etc.
(4) Certification. Certification of authenticity by the study director
(including signature, typed name, title, affiliation, address, telephone num-
ber, date).
(5) Tables and figures.
(6) References.
(7) Appendices. Include: Representative chromatograms, spectra, etc.
(as applicable) and any relevant material not fitting in any of the other
sections to this report.
(e) References. The following references should be consulted for ad-
ditional background material on this test guideline.
(1) Association of Official Analytical Chemists (AOAC) Inter-
national, Official Methods of Analysis, latest edition. Available from
AOAC International, 2200 Wilson Boulevard, Arlington, VA 22201.
(2) Environmental Protection Agency, EPA Test Methods for Evaluat-
ing Solid Waste (SW-846), Method 8280 and EPA Test Methods for Or-
ganic Chemical Analysis, Method 613.
(3) Environmental Protection Agency, PR Notice 96-1, Tolerance En-
forcement Methods—Independent Laboratory Validation by Petitioner,
Feb. 7, 1996.
(4) FEDERAL REGISTER 58:50888-50893, Sept. 29, 1993.
(5) Horwitz, W. et al, Quality Assurance in the Analysis of Foods
for Trace Constituents, JAOAC, Vol. 63, No. 6, pp. 1344-1354 (1980).
(6) Pesticide Analytical Manual. Food and Drug Administration. Vol-
ume I. (1994). Available from the National Technical Information Service,
Springfield, VA.
(7) Pesticide Analytical Manual. Food and Drug Administration. Vol-
ume II. (1994). Available from the National Technical Information Serv-
ice, Springfield, VA.
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