a EPA
United States
Environmental Protection
Agency
Office of Chemical Safety and
Pollution Prevention (7101)
EPA 712-C-10-001
July 7, 2010
Product Performance
Test Guidelines
OPPTS 810.3700:
Insect Repellents to be
Applied to Human Skin
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NOTICE
This is one of a series of test guidelines established by the Office of Chemical Safety and
Pollution Prevention (OCSPP) (formerly the Office of Prevention, Pesticides and Toxic
Substances (OPPTS) prior to April 22, 2010),United States Environmental Protection Agency for
use in testing pesticides and chemical substances to develop data for submission to the Agency under
the Toxic Substances Control Act (TSCA) (15 U.S.C. 2601, et seq.\ the Federal Insecticide, Fungicide
and Rodenticide Act (FIFRA) (7 U.S.C. 136, etseq.), and section 408 of the Federal Food, Drug and
Cosmetic Act (FFDCA) (21 U.S.C. 346a), referred to hereinafter as the harmonized test
guidelines.
The harmonized test guidelines serve as a compendium of accepted scientific methods for
research intended to provide data to inform regulatory decisions under TSCA, FIFRA, and/or FFDCA.
This document provides guidance for conducting appropriate tests, and is also used by EPA, the
public, and the companies that are required to submit data under FIFRA. These guidelines are not
binding on either EPA or any outside parties, and EPA may depart from them where circumstances
warrant and without prior notice. The methods described in these guidelines are strongly
recommended for generating the data that are the subject of the guidelines, but EPA recognizes that
departures may sometimes be appropriate. You may propose and alternatives to the methods described
in these guidelines, with your supporting rationale. The Agency will assess such proposals on a case-
by-case basis.
For additional information about the harmonized test guidelines and to access the guidelines
electronically, please go to http://www.epa.gov/oppts and select "Test Methods & Guidelines" from
the navigation menu at the top of the screen. You may also access the guidelines in
http://www.regulations.gov grouped by Series under Docket ID #s: EPA-HQ-OPPT-2009-0150
through EPA-HQ-OPPT-2009-0159, and EPA-HQ-OPPT-2009-0576.
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OPPTS 810.3700: Insect Repellents to be Applied to Human Skin
(a) Introduction.
(1) Scope and Purpose. This guideline provides recommendations for the design
and execution of studies to evaluate the performance of pesticide products
intended to repel insects and other arthropods in connection with the products'
registration under the Federal Insecticide, Fungicide, and Rodenticide Act
(FIFRA) (7 U.S.C. 136, et seq.). This guidance applies to products in any
formulation—such as lotion, liquid, or spray—intended to be applied directly to
human skin. It does not apply to products applied to or impregnated into clothing
or fabric, or used to repel insects from indoor or outdoor spaces. This guidance
recommends appropriate study designs and methods for selecting subjects,
statistical analysis, and reporting.
(2) General Considerations. Any protocol developed using this guidance must meet
the requirements set forth in several statutes and regulations, including, but not
limited to, the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA, 7
U.S.C. 136, et seq.) under which EPA regulates repellents, and EPA's rules for
the protection of human subjects of research, 40 CFR part 26, subparts K through
Q. Because these studies would support a FIFRA registration and involve
intentional exposure of human subjects to the test repellents, review of each
protocol by an Institutional Review Board (IRB), by EPA, and by EPA's Human
Studies Review Board (HSRB) is required before the study is initiated. (See 40
CFR §26.1109, §26.1125, and §26.1601.)
This guideline does not supersede or overrule the regulations governing research
conducted on human subjects contained in 40 CFR part 26, subparts K through Q,
or any other Agency regulations. To the extent there are any unintended conflicts
between this guideline and any EPA regulation, the regulation at issue governs.
(3) Related Requirements.
(i) FIFRA Informed Consent Requirement. Any research conducted under
this guideline is subject to §12(a)(2)(P) of FIFRA, which defines it as an
unlawful act for any person "to use any pesticide in tests on human beings
unless such human beings (i) are fully informed of the nature and purposes
of the test and of any physical and mental health consequences which are
reasonably foreseeable therefrom, and (ii) freely volunteer to participate in
the test." Regulations implementing this statutory provision and defining
associated record-keeping requirements can be found at 40 CFR §169.2(j)
and (k).
(ii) EPA's Rule for the Protection of Human Subjects of Research. Any
research conducted under this guideline is covered by the requirements of
EPA regulations for the protection of human subjects of research set out at
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40 CFR part 26, subparts K, L, and M. Persons conducting and submitting
topical repellent efficacy studies should ensure compliance with all
applicable requirements of that rule; the following paragraphs highlight a
few of them:
(A) Applicability. Subparts K and L of 40 CFR part 26 apply to
regulated third parties who conduct or sponsor research involving
intentional exposure of human subjects which is intended for
submission to EPA for consideration under the pesticide laws—
FIFRA and §408 of the Federal Food, Drug, and Cosmetic Act
(FFDCA) (21 U.S.C. 346a). (40 CFR §26.1101) Efficacy testing
of topically applied repellents typically meets the regulatory
definitions of "research" with "human subjects" involving
"intentional exposure," and must therefore be conducted and
submitted to EPA in compliance with the requirements of these
subparts. (See definitions at 40 CFR §26.1102.) Subpart M
applies to any person who submits data from research with human
subjects for consideration in connection with any action that may
be performed by EPA under FIFRA or section 408 of FFDCA.
(B) Prohibition of research involving pregnant or nursing women
or children. 40 CFR §26.1203 as amended effective August 22,
2006, provides:
"Under no circumstances shall a person conduct or sponsor
[covered] research that involves intentional exposure of any human
subject who is a pregnant woman (and therefore her fetus), a
nursing woman, or a child."
(C) Required review by Institutional Review Board (IRB). 40 CFR
§26.1109(a) provides:
"An IRB shall review and have authority to approve, require
modifications in (to secure approval), or disapprove all research
activities covered by this subpart."
(D) Required Pre-testing submissions to EPA. 40 CFR §26 1125
provides:
"Any person or institution who intends to conduct or sponsor
human research covered by §26.1101(a) shall, after receiving
approval from all appropriate Institutional Review Boards (IRBs),
submit to EPA prior to initiating such research all information
relevant to the proposed research specified by §26.1115(a), and, to
the extent not already included:
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(a) A discussion of:
(1) The potential risks to human subjects;
(2) The measures proposed to minimize risks to the
human subjects;
(3) The nature and magnitude of all expected benefits
of such research, and to whom they would accrue;
(4) Alternative means of obtaining information
comparable to what would be collected through the
proposed research; and
(5) The balance of risks and benefits of the proposed
research.
(b) All information for subjects and written informed consent
agreements as originally provided to the 1KB, and as
approved by the 1KB.
(c) Information about how subjects will be recruited, including
any advertisements proposed to be used.
(d) A description of the circumstances and methods proposed
for presenting information to potential human subjects for
the purpose of obtaining their informed consent.
(e) All correspondence between the 1KB and the investigators
or sponsors.
(f) Official notification to the sponsor or investigator, in
accordance with the requirements of this subpart, that
research involving human subjects has been reviewed and
approved by an 1KB."
(E) Required post-testing submissions to EPA. 40 CFR §26 1303
provides:
"Any person who submits to EPA data derived from human
research covered by this subpart shall provide at the time of
submission information concerning the ethical conduct of such
research. To the extent available to the submitter and not
previously provided to EPA, such information should include:
(a) Copies of all of the records relevant to the research
specified by §26.1115(a) to be prepared and maintained
by an 1KB.
(b) Copies of all of the records relevant to the information
identified in §26.1125(a) through (f).
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(c) Copies of sample records used to document informed
consent as specified by §26.1117, but not identifying
any subjects of the research."
(F) Prohibition of EPA reliance on unethical human research. 40
CFR §26.1705 provides:
"... EPA shall not rely on data from any research initiated after
April 7, 2006, unless EPA has adequate information to determine
that the research was conducted in substantial compliance with
subparts A through L of this part, or if conducted in a foreign
country, under procedures at least as protective as those in subparts
A through L of this part."
(iii) Good Laboratory Practice Standards. Good Laboratory Practice
Standards (GLP) as defined in 40 CFR part 160 apply to both laboratory
and field studies of repellent efficacy. According to 40 CFR §160.17:
"EPA may refuse to consider reliable for purposes of supporting an
application for a research or marketing permit any data from a study which
was not conducted in accordance with this part." 40 CFR §160.12(b)
requires with any submitted research data "[a] statement describing in
detail all differences between the practices used in the study and those
required by this part." Additionally, 40 CFR part 158 specifies that
"applicants must adhere to the good laboratory practice (GLP) standards
described in 40 CFR part 160 when conducting studies." (40 CFR
§158.70(b)).
(iv) State Requirements. Investigators and Sponsors should ensure research
is conducted in compliance with any applicable state laws or regulations,
which are independent of and additional to those cited here.
(4) Organization of the Guideline. This guideline begins with definitions of special
importance in understanding this guideline (Section b). Sections (c) through (h)
provide general guidance applicable to all topical repellent efficacy testing,
whether conducted in the laboratory or in the field, and to all target species. Each
of these six sections discusses one of the primary stages of repellent testing.
(c) Development of protocols for repellent studies.
(1) Scientific design of repellent studies.
(2) Ethical justification for repellent studies.
(3) Subject selection and informed consent.
(4) Protection of subject privacy and confidentiality
(d) Review of protocols for repellent studies.
(e) Changes to IRB-approved research before execution.
(f) Execution of repellent studies.
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(g) Reporting of completed repellent studies.
(h) Retention of records.
Sections (i) through (1) provide guidance specific to different kinds of testing of
repellents. Section (i) addresses empirical estimation of a typical consumer dose,
and sections (j) through (m) address testing of repellency in the laboratory or field
to particular species of arthropods.
(i) Specific guidance for dose-determination studies.
(j) Specific guidance for laboratory studies of mosquito or biting fly
repellency.
(k) Specific guidance for field studies of mosquito or biting fly repellency.
(1) Specific guidance for laboratory studies of tick or chigger repellency.
The final section (m) lists references considered in the development of this
guideline.
Three appendices are attached as well:
Appendix A: Checklist of Elements Required by 40 CFR §26.1125.
Appendix B: Framework for Science and Ethics Reviews of Proposed
Human Research.
Appendix C: Checklist of Elements Required by 40 CFR §26.1303.
(b) Definitions. The following definitions are of special importance in understanding this
guideline. They apply only in the context of this guideline and are not intended to be
more generally applicable.
(1) The following events may indicate a failure of repellent efficacy.
(i) A landing is the act of a flying or jumping insect or other arthropod
alighting on human skin without probing or biting.
(ii) A probe is the act of penetrating human skin by the mouthparts of an
insect or other arthropod without ingestion of blood.
(iii) A bite is the act of penetrating human skin by the mouthparts of an insect
or other arthropod with ingestion of blood, typically associated with
abdominal swelling and color change.
(iv) A crossing is the act of passage by a tick or chigger from an area of
untreated skin to an area of treated skin. A crossing may be quantified
either or both by the distance the tick or chigger moves onto treated skin
or by how long the tick or chigger remains on treated skin.
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(2) An unconfirmed event is a landing, probe, bite, or crossing not followed by
another similar event within 30 minutes.
(3) A confirmed event is one landing, probe, bite, or crossing followed by another
similar event within 30 minutes. The first event is confirmed by the second; the
second event is the confirming event.
(4) A human subject is a living individual about whom an investigator conducting
research obtains either data through intervention or interaction with the individual
or identifiable private information. By this definition, both untreated control
subjects and treated subjects are considered human subjects of repellent efficacy
testing.
(5) Questing is the behavior of ticks or chiggers actively seeking a host.
(6) A repellent is a product intended to disrupt the host-seeking behavior of insects or
other arthropods, driving or keeping them away from treated human skin.
(7) Complete Protection Time (CPT) is the time from application of a repellent until
efficacy failure as it is defined in each study—for example, the time from
application until the first efficacy failure event confirmed within 30 minutes by a
second similar event.
(8) Dose determination is a testing procedure used to estimate a "typical consumer
dose" of a topical repellent.
(c) Development of protocols for repellent studies. The first major stage of repellent
testing is development of a protocol. Under EPA's amended rules for the protection of
human subjects—40 CFR part 26—protocol development is the focus of far more
attention than was the case before that rule took effect in April 2006. A detailed protocol
must be approved by an Institutional Review Board (IRB) and, accompanied by
supporting documentation meeting the requirements of 40 CFR §26.1125, must then be
reviewed by EPA and the HSRB before research is initiated.
It is critical to all later stages of repellent testing that protocols contain the elements
required by EPA's Good Laboratory Practices regulations at 40 CFR §160.120 and meet
all other applicable scientific and ethical standards. The broad topics of scientific design,
ethical justification, subject selection and informed consent, and protection of subject
privacy and confidentiality are discussed in detail below.
(1) Scientific Design of Research. To be scientifically justified, the proposed
research should address an important research question that cannot be answered
by existing data. In addition, the design should be such as to be likely to provide
a definitive answer to the research question. The design should include a detailed
description of the experimental design, addressing topics (i) through (x).
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(i) Objectives. The objective of most repellent efficacy testing to support
registration is to estimate how long after treatment a repellent will
continue to protect users from the target pest; this period of effective
repellency is measured as CPT. Sometimes the objective of repellent
efficacy testing is to compare the efficacy of one treatment to another
treatment or to an untreated control; this ratio is expressed as Relative
Protection (RP). In dose-determination testing associated with repellent
testing the objective is typically to estimate a "typical consumer dose" for
use in later repellent testing. In all cases the scientific objective should be
stated clearly.
(ii) Test materials. Repellent efficacy should be tested using the end-use
formulation as registered or as proposed for registration. Test materials
should be stored at ambient temperature and humidity before use.
(iii) Choice of endpoints and measures. Endpoints chosen for the study
should be appropriate for the specific objectives of proposed research and
likely to provide a robust answer to the research question while
minimizing the risks to subjects. Considerations in endpoint selection for
dose determination and repellency include:
(A) Dose determination. To estimate a "typical consumer dose" the
endpoint of concern is a rate of application, typically expressed
either by weight as mg/cm2 of treated skin surface or
volumetrically as ml/cm2 of treated skin surface. Because each
subject is likely to apply a different amount of repellent in
uncontrolled trials, the "typical dose" should be calculated as the
mean of multiple applications by each of many subjects.
(B) Repellency. The endpoint of repellency testing should be selected
to show a failure of repellent efficacy for subjects treated with a
"typical consumer dose." Efficacy failure in a test to determine
Complete Protection Time (CPT) may be defined either as the first
event—i.e., the first landing, bite, or crossing—or as the first
confirmed event—i.e., the first landing, bite, or crossing confirmed
within 30 minutes by another similar event. The Agency
encourages the use of the first confirmed failure event to estimate
complete protection time for individual subjects, and the median
CPT across all subjects in each treatment arm as the summary
measure of CPT.
(7) In repellency testing with mosquitoes or biting flies the
study design should choose either landings or bites as the
event showing failure of repellent efficacy. The Agency
encourages the choice of landings for field testing to reduce
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the risk of exposing subjects to vector-borne diseases.
Even in laboratory testing using laboratory-reared, disease-
free insects, the choice of landings reduces the risk of an
adverse reaction to bites. A proposal to use bites as an
endpoint in either field or laboratory testing should be
justified.
(2) In repellency testing with ticks or chiggers the appropriate
event to show failure of repellent efficacy is a "crossing"
from untreated skin onto treated skin.
(iv) Duration. Repellency testing should continue long enough to fairly
assess the duration of protection provided by the repellent, and long
enough that all or nearly all subjects experience efficacy failure.
If testing ends before some subjects experience a failure of efficacy the
resulting data-set is said to be "right-censored." Right-censorship of data
poses challenges for meaningful and accurate analysis. Right-censorship
can be reduced through appropriate care in study design—for example, by
planning a test of longer duration or, when appropriate, by pre-treating
subjects well before their first exposure to target pests. Methods for
analyzing right-censored datasets are discussed in paragraph (c)(l)(xi)
below.
(v) Sample size. The sample should be large enough to be likely to yield a
definitive answer to the research question being addressed, and its size
should be justified statistically in each protocol, taking into account the
specific characteristics of the proposed research and the desired accuracy
and precision of the results. Researchers are encouraged to consult a
statistician to help determine appropriate sample size.
Withdrawal of test subjects from the study before failure of efficacy
decreases the sample size and may compromise the validity or utility of
test results. The protocol should fully describe how the proposed sample
size was determined, and how data for subjects who withdraw from the
test prematurely will be treated.
Other factors which may affect sample size are the number of treatments,
the experimental design, and the heterogeneity of the target population
(e.g., by age, sex) and of the environment (different habitats, different
conditions, different species and population densities).
(vi) Allocation of subjects to treatments. Subjects should be allocated to
treatments randomly and, to the extent possible, blindly.
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Multiple treatments per subject may be permissible if all treatments
contain the same active ingredient at comparable concentrations, but care
must be taken in the study design and execution to prevent cross-
contamination or confounding interference. It is particularly important for
subjects receiving different treatments on different limbs not to rub their
limbs together or otherwise contaminate the repellent treatments, to keep
results independent. The number of treatments per subject should be
limited by the feeding behavior of the target species. For example, for
mosquito species feeding close to the ground on lower limbs, treatments
should be applied only to subjects' lower legs, and only two treatments per
subject will be possible. Treating the same subject more than once with
the same treatment does not increase the sample size; a single subject can
only be counted once in determining the sample size.
When assessing relative protection provided by more than one product
tested simultaneously, a Latin square design, in which each subject is
tested with each treatment over the course of the study, is often
appropriate.
(vii) Untreated controls. To minimize risks to untreated control subjects, both
the number of untreated subjects and the duration of their exposure should
be minimized. Untreated controls should be exposed intermittently, and
then only long enough to confirm adequate pest pressure throughout the
study. Recent research (Barnard, et al. 2002) has shown that treated
subjects should not serve as their own untreated controls.
(viii) Positive controls. Positive controls are desirable to support comparison
of results from different repellency studies or from testing on different
days or at different locations. The recommended positive control material
is 20% deet in ethanol, applied at a standard dose rate of 1 ml per 600 cm2.
Positive controls are particularly valuable when the objectives of the study
include comparisons between formulations, and when testing spans
different days. To minimize the likelihood of interference, subjects treated
with the test material should not also serve as concurrent positive controls.
(ix) Preparation of subjects. Before treatment with a test repellent or use as
an untreated or positive control, subjects' limbs should be washed with an
unscented detergent and carefully rinsed and dried. Subjects should avoid
alcohol, tobacco, and scented products (perfume, cologne, hair spray,
lotion, soap, etc.) for at least twelve hours before and throughout the test.
Subjects should avoid activities that increase perspiration, and avoid
abrading, rubbing, touching, or wetting the treated area.
(x) Treatment of subjects. Subjects in trials designed to estimate a typical
consumer dose should self-treat with the repellent, which should be
provided in the type of container and delivery system (e.g., pump spray,
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aerosol spray, towelette, or lotion) and with the directions for use intended
for commercial distribution. Subjects in repellency trials should all be
treated at the same standard dose rate, and treatment should be delivered
so as to ensure consistent application and uniform coverage.
(xi) Statistical analysis plan. Protocols should include a full description,
explanation, and justification for the statistical methods proposed to
analyze both dose determination and repellency test results, taking into
account the specific study objectives and variables.
The statistical analysis plan should provide for testing results for normal
distribution. When results are normally distributed, it may be appropriate
to report the mean CPT across all treated subjects with its standard error.
When the data do not fit a normal distribution—more typical of repellency
datasets—it may be possible to transform them to fit a distribution for
which a parametric method of analysis can be employed. When the data
do not fit and cannot be transformed to fit an underlying distribution, non-
parametric analyses, such as Kaplan-Meier survival analysis1, are
suggested.
If the study objective is to determine CPT across all subjects, any right-
censorship of repellency data will lead to underestimation of both the
mean and the variance around it. Because right-censorship is common in
repellency testing, EPA recommends use of the median CPT with its 95%
confidence limits as the summary measure of CPT.
(xii) QA/QC plan. Protocols should provide for periodic quality assurance
inspections adequate to ensure the integrity of the study and consistent
with the requirements of EPA's Good Laboratory Practices regulations (40
CFR part 160.)
(2) Ethical Justification for Research. Because repellent efficacy testing is never
directly beneficial to the subjects of the research, the risks to subjects must be
minimized for the research to be ethically justified. The expected benefits to
society from the knowledge likely to be gained in the research must also outweigh
the minimized risks to subjects. (40 CFR §26.1111)
(i) Alternatives to research with human subjects. If the research question
can be answered without conducting new research with human subjects,
human research is not justifiable. Investigators should consider possible
alternatives to human research; protocols should discuss possible
alternatives to human research and explain why they are infeasible or
would not answer the research question.
1 For guidance see, for example, WHO (2009) Guidelines for Efficacy Testing of Mosquito Repellents for Human
Skin: WHO/HTM/NTD/WHOPES/2009.A. Annex 3: Estimation of Median and Confidence Interval of Complete
Protection Time Using the Kaplan-Meier Survivor Function.
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(ii) Prerequisite research. Before testing a repellent's efficacy with human
subjects, acute toxicity to animals of the active ingredients should be
tested and reported in the protocol. Acute dermal toxicity, dermal
irritation, eye irritation, and skin sensitization studies are needed to
estimate the margin of exposure (MOE) for subjects participating in a
repellent test. All additional components in the materials tested should be
cleared by EPA for use in repellent formulations.
(iii) Risks to subjects. Protocols should include a complete discussion of risks
to subjects associated with their participation in the research,
characterizing all risks and their likelihood, and describing steps proposed
to minimize each of them.
(A) Risk characterization. The nature of all risks to subjects should
be described and the probabilities of occurrence of each type of
risk should be estimated in the protocol. Potential risks in repellent
testing typically include (but may not be limited to) reactions to the
test substance or to arthropod bites, acquisition of vector-borne
illness, physical risks (e.g., stresses imposed by the requirements
or conditions of the test), or possible psychological risk associated
with a breach of confidentiality in handling the results of
pregnancy testing.
(B) Risk minimization. As a condition of approval of proposed
research, an 1KB must determine that risks to subjects have been
minimized by using procedures which are consistent with sound
research design and which do not unnecessarily expose subjects to
risk. (See 40 CFR §26.111 l(a)(l).) Different actions are effective
in minimizing different risks; protocols should identify specific
steps taken to minimize each identified risk to subjects.
Protocols should discuss specific steps proposed to prevent
research-induced harm—for example, by training subjects, using
appropriate eligibility criteria, ensuring adequate supervision of
subjects, and having a physician on call during testing. In addition,
protocols should specify how any research-induced harm will be
reversed—for example, by the investigator's or sponsor's
committing to pay for uninsured costs of medical treatment of
subjects for injuries or illnesses resulting from their participation in
the research, and for longer-term treatment, if needed, of any
arthropod-borne disease contracted during testing.
Study protocols should include procedures for monitoring the
safety of subjects, stopping rules specifying conditions under
which a subject would be withdrawn from the research or the
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research would be terminated to protect subjects, and a medical
management plan covering foreseeable contingencies.
Methods of risk minimization are ultimately study-specific, but
ensuring that risks to subjects are minimized in repellent testing
may involve some or all of the following:
• Monitoring of potential field testing sites for the presence
of disease vectors at least weekly during the month before
testing. Sites at which infected vectors are known to be
present should not be used for field testing;
• Serological or DNA-based assays of insects collected at the
site of field testing to determine the presence or absence of
disease organisms. Results of such testing should be
reported to subjects after completion of the field test;
• Using the minimum number of untreated controls
consistent with statistical soundness;
• Exposing untreated control subjects intermittently for only
the minimum time required to confirm continued pest
pressure;
• Training subjects to use an aspirator to capture landing
insects before they have time to bite;
• Using only pathogen-free laboratory-reared insects in
laboratory tests;
• Excluding subjects known to be sensitive to repellents or to
insect bites;
• Using bites as the endpoint in either field or laboratory
studies only when fully justified;
• Ensuring that enough sub-investigators are present at all
times so that the Principal Investigator can, if needed,
attend to the safety of a subject without compromising the
integrity of the research or endangering other subjects;
• Initiating post-study contact with subjects to inquire about
any signs of study-related injury or illness.
(iv) Expected benefits of research. Participation as a subject in repellent
testing has no direct benefit for individual subjects, yet is not risk-free,
even after risks have been minimized. Thus the ethical justification for the
research ultimately depends on the expected benefits to society of the
information likely to be gained from the research. Protocols should
characterize all anticipated benefits of the information to be gained from
the research, and to whom or to what segment in society each identified
benefit is likely to accrue. In addition, protocols should estimate the
likelihood of achieving each identified societal benefit.
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Societal benefits include only those directly resulting from the knowledge
likely to be gained from the research. Benefits potentially resulting from
use of effective repellents should not be attributed to the proposed
research. Payments to subjects or other incentives for participation should
not be treated as benefits, either to the subjects or to society.
(v) Relation of benefits to risks. As a condition of approval of proposed
research, an 1KB must also determine that minimized "risks to subjects are
reasonable in relation to anticipated benefits, if any, to subjects, and the
importance of the knowledge that may reasonably be expected to result. In
evaluating risks and benefits, the 1KB should consider only those risks and
benefits that may result from the research. . . . The 1KB should not
consider possible long-range effects of applying knowledge gained in the
research ... [to be] within the purview of its responsibility." (See 40 CFR
§26.111 l(a)(2).) The protocol should discuss explicitly the balance of
risks to subjects and anticipated societal benefits of the research.
(3) Subject selection and informed consent. Subject selection is important both to
the scientific merit of research and to its ethical acceptability. Selection of
representative subjects is critical to the generalizability of the results of research,
and to be ethically acceptable, subject selection must be equitable. (40 CFR
§26.111 l(a)(3)) Fully informed and fully voluntary consent of subjects is
fundamental to ethical human research. (FIFRA §12(a)(2)(P) and 40 CFR
§26.1116)
(i) Representative sampling. The results of repellent testing should be as
generalizable as possible to the target population of repellent users.
Samples should include adults of various ages, of both sexes, and of a
variety of races and ethnicities, and protocols should describe the
demographic characteristics of the pool from which subjects will be
recruited. Attractiveness of recruited candidates to the target species
should be verified before they participate in repellency testing.
Testing with a sample known to be unrepresentative is discouraged; if it is
proposed, it should be fully justified. Arguments based on the
convenience of the investigators or on the difficulties associated with
recruiting a more representative sample do not justify testing with an
unrepresentative sample.
(ii) Inclusion/Exclusion factors. All exclusion/inclusion criteria proposed
should be supported by a clear and explicit rationale. Because testing a
topical repellent necessarily involves intentional exposure of subjects,
children under 18 and pregnant or nursing women must be excluded. (See
40 CFR §26.1203.) Because older adults are more susceptible to
arthropod-borne diseases, candidates over 55 should be excluded as
subjects in field testing, or their inclusion should be specifically justified.
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Members of certain vulnerable populations, including people of limited
mental capacity, those not in good health or with compromised immune
systems, those sensitive to chemicals, and students or employees of the
investigators or sponsors should always be excluded. Other potential
subjects who may be in a vulnerable position, including the educationally
or economically disadvantaged, or those who communicate with difficulty
because of language differences or disabilities, should not be excluded
arbitrarily if appropriate specific provision can be made to ensure their
safely and welfare.
Subjects should generally be recruited from populations in the area where
testing will be conducted. An offering of distant travel may unduly
influence a candidate's choice to enroll, and a subject who has accepted
long-distance transportation may feel less than free to withdraw from a
study. If it is proposed, transportation of subjects to distant locations
should be justified, and specific mechanisms should be proposed to
prevent any coercion or undue influence.
(iii) Methods of recruiting. Protocols should describe in detail the proposed
recruiting process, from the first contacts with potential candidates
through all discussions of the research and the subjects' enrollment. Any
advertisements or flyers proposed for use in recruiting should be appended
to the protocol, and should reflect IRB review and approval. If recruiting
will be done through telephone calls, the script for recruiting calls should
be appended to the protocol, and should reflect IRB review and approval.
If any candidates may prefer to speak or read a language other than
English, procedures for accommodating their needs in the recruiting
process and in the conduct of the research itself should be described in
detail.
(iv) Compensation of subjects. It is reasonable to compensate participants in
repellent efficacy research for their time and trouble. The level of
compensation should not be so high as to constitute an undue influence in
the choice to participate, nor should it be so low as to make participation
in the research attractive only to the economically disadvantaged.
Compensation should not be used or administered so as to compromise the
freedom guaranteed to subjects to withdraw from participation at any time
for any reason, without sacrificing benefits to which they are entitled.
(v) Informed consent. It is a fundamental requirement for ethical research
with human subjects that participation of subjects be both fully informed
and fully voluntary.
(A) Process. Informed consent is an extended process, involving
much more than simply obtaining a subject's signature on a form.
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It begins with recruiting, and continues after signature of the
consent form throughout the conduct of the research. A single
discussion in the protocol of the recruiting and consent processes
as a continuous whole is recommended.
As a condition of approval of proposed research, an Institutional
Review Board (1KB) must determine that informed consent will be
sought from each prospective subject in accordance with, and to
the extent required by §26.1116, and that informed consent will be
appropriately documented, in accordance with and to the extent
required by §26.1117. (See 40 CFR §26.111 l(a)(4) and (5).)
The rule at §26.1116 requires that consent be sought "only under
circumstances that provide the prospective subject. . . sufficient
opportunity to consider whether or not to participate and that
minimize the possibility of coercion or undue influence. The
information that is given to the subject shall be in language
understandable to the subject." EPA recommends that consent
discussions with individual candidates be conducted in private, and
that all consent documents be written at or below an 8th grade
reading level, as measured by the Flesch-Kincaid Grade Level
score or another standard tool for assessing readability. (A tool to
calculate the Flesch-Kincaid Grade Level score is a standard
feature available to users of MS-Word.) If consent materials are
translated into another language, it is important to ensure the
translations are also at an appropriate reading level.
The informed consent process should describe how the investigator
will confirm each candidate's understanding of the research and of
what it will involve before he or she is enrolled.
The rule at §26.1116 further defines the general requirements for
informed consent, including at §26.1116(a) the basic elements
always required, at §26.1116(b) other elements conditionally
required, and at §26.116(e) a requirement to identify the pesticide
and the nature of its pesticidal function.
(B) Documentation of informed consent. The rule at §26.1117
provides two options for documenting informed consent. The first
option is a "written consent document that embodies the elements
of informed consent required by § 26.1116." The second is "a
short form written consent document stating that the elements of
informed consent required by § 26.1116 have been presented orally
to the subject." The short form requires that a witness be present
in the consent interview. In repellent efficacy testing the short
form is generally inappropriate; if use of a short form is proposed,
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it should be justified, and prior discussion with EPA is strongly
encouraged.
Ensure that any elements appearing in both the protocol and the
consent form are in harmony. Elements common to both
documents may include, for example, descriptions of the purpose
of the research, subject inclusion/exclusion factors, risks to
subjects and how they will be minimized, and what will happen in
the course of the research. The protocol and the consent form
should be consistent in substance, although the presentations
should usually not be the same, because the readers and purposes
differ. Consent forms should address candidates consistently in the
second person—as "you"—and should present all information
from the subject's point of view. By contrast protocols should be
written in the third person, and should present all information from
the investigators' point of view.
In repellent testing, the consent form should tell subjects how
many bites, if any, they are likely to get, and what symptoms of
arthropod-borne disease they should be alert for after participating
in a field study.
(4) Protection of subject privacy and confidentiality. It is important to protect the
privacy and confidentiality of subjects in repellent efficacy research, and to
inform candidates in the recruiting and consent processes of the extent, if any, to
which confidentiality of records identifying them will be maintained.
(i) Subject identification. Subjects should not be identified by name in data
collection forms or study reports. Subject names unavoidably must appear
on consent forms and on administrative documents, but subjects should be
identified only by an arbitrary code on other study documents. The key
linking identifying codes to subjects should be stored securely, away from
other study records.
(ii) Photographic images. If photographs or videos are made to document
the research, care should be taken to minimize making recognizable
images of subjects. If faces or identifying marks cannot be excluded from
a photograph or video image, the image should be altered to protect the
identity of the subject(s).
(iii) Pregnancy testing. Testing of female candidates for pregnancy should be
handled with care and discretion. The investigator is responsible for
ensuring that pregnant or nursing women are excluded as subjects (See 40
CFR §26.1203). It is not necessary to meet this responsibility by testing
all female candidates for pregnancy. For example, it is neither respectful
nor informative to require women who are post-menopausal or surgically
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incapable of pregnancy to take a pregnancy test. Investigators should
consider how best to meet their responsibilities while fully respecting
female subjects.
When pregnancy testing is conducted, the circumstances of testing should
protect the privacy of the candidate, both during the testing itself and in
handling the results. Unexpected news of a pregnancy can cause
significant psychological distress, which can be increased by any breach
of discretion in handling the information. It is a good practice to recruit
more candidates than are required by the study design, so that the design
would not be compromised by withdrawal of one or more subjects. This
approach permits a candidate with a positive pregnancy test to withdraw
without stating a reason. No records of a positive pregnancy test should
be retained.
(d) Review of protocols for repellent studies. EPA's Rule for the Protection of Human
Subjects of Research requires extensive review of protocols before research is initiated.
The rule requires that the complete protocol, consent form, and supporting materials be
reviewed and approved by an IRB (See 40 CFR §26.1109), and having been approved by
the IRB, that it be submitted, along with specified additional supporting documentation,
to EPA for review by EPA and by the HSRB (See 40 CFR §26.1125 and §26.1601.)
(1) Review by IRB. The membership, functions, general procedures, and decision
criteria for IRBs are defined in the rule at 40 CFR §26.1107-§26.1111. Records
to be maintained by IRBs are defined at 40 CFR §26.1115. Most IRBs review
many other kinds of human research in addition to repellent efficacy studies.
Extensive guidance to and about IRBs can be found on the website of the Office
of Human Research Protections (OHRP), at www.hhs.gov/ohrp.
(i) Required elements in submission to IRB. Each IRB has its own
application forms and procedures. Typical requirements are for a
complete protocol, a consent form, and all materials intended for use in the
recruiting process or to be provided to candidates or subjects. A protocol
responsive to section (c) of this guideline is likely to satisfactorily address
most requirements of a reviewing IRB.
(ii) Criteria for IRB approval. The criteria for IRB approval of proposed
research are defined in the rule at 40 CFR §26.1111. In summary, this
passage of the rule requires that the IRB determine that proposed research
meets all these criteria:
• Risks to subjects are minimized;
• Risks to subjects are reasonable in relation to the importance of the
knowledge that may be reasonably expected to result from the
research;
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• Selection of subjects is equitable;
• Informed consent will be sought from each prospective subject;
• Informed consent will be appropriately documented;
• When appropriate, adequate provision is made for monitoring
collected data to ensure the safety of subjects; and
• When appropriate, adequate provision is made to protect the
privacy of subjects and to maintain the confidentiality of data.
In addition, the 1KB ensures that when any subjects are likely to be
vulnerable to coercion or undue influence, additional safeguards are
included in the study design to protect their rights and welfare.
(iii) Documentation of 1KB approval. IRBs have the authority to approve,
require modifications in (to secure approval), or disapprove research
proposals. (40 CFR §26.1109.) It is common for IRBs to grant
conditional approval subject to the investigator's making specified
modifications in the proposal; once required modifications are made, the
IRB will issue formal written approval of the research. This letter of
approval, along with all other correspondence between the investigators or
sponsors and the reviewing IRB, must be included in the subsequent
submission to EPA. (See 40 CFR § 26.1125(e) and (f).)
(2) Reviews by EPA and the HSRB. After receiving approval from an IRB, the
protocol and supporting material must be submitted to EPA for review by EPA
and by the HSRB. (See 40 CFR § 26.1125.)
(i) Required elements in submission to EPA. The elements to be included
in a protocol submission to EPA are defined in the rule at 40 CFR
§26.1125 and summarized in Appendix A to this guideline. Records of
two kinds are required.
(A) Records describing the research proposal itself. 40 CFR
sections 26.1125(a)-(d) call for a discussion of the risks and
benefits of the proposed research, for the consent forms as
submitted to and as approved by the IRB, for information about
how subjects will be recruited, and for a description of the
circumstances and methods proposed for presenting information to
subjects and seeking their consent. A protocol responsive to the
guidance in section (c) of this guideline will already include this
range of information.
In addition, 40 CFR §26.1125(e) and (f) call for submission of all
correspondence between the IRB and the investigators or sponsors
(§26.1125(e)) and official notification of IRB review and approval
(§26.1125(f)). For purposes of §26.1125(e), "correspondence"
does not include attachments transmitted by correspondence, such
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as the protocol or consent form, which are otherwise required to be
submitted by other provisions of the rule.
(B) Records of the 1KB review. 40 CFR §26.1125 also requires
submission of copies of records relevant to the proposed research
which the 1KB is required to maintain by 40 CFR §26.1108(a).
There is some overlap between these records and those discussed
above, but there are some important differences. 40 CFR
§26.1108(a) calls for, among other records:
• Minutes of IRB meetings, in sufficient detail to show
attendance; actions taken; votes for, against, and
abstaining; the basis for requiring changes, and
summarizing the discussion of controverted issues;
• A list of IRB members identified by name, earned degrees,
representative capacity, indications of experience sufficient
to describe each member's chief anticipated contributions
to IRB deliberations, and any employment or other
relationship between each member and the research
institution; and
• Written procedures for the IRB in the same detail described
in §26.1108(a) and §26.1108(b).
These records can only be obtained from the IRB. By prior
arrangement, however, an IRB may submit them directly to EPA.
(ii) EPA Review. Timely EPA review of submitted proposals for research is required
by the rule at 40 CFR §26.1601. EPA will first review submitted proposals for
satisfaction of the requirements of 40 CFR §26.1125, and for substantiation of any
claims of business confidentiality associated with the submission. If a submission
is found to be incomplete, or to include unsubstantiated claims of business
confidentiality, the submitter will be promptly notified; the submission will not be
reviewed further until these deficiencies are corrected. EPA evaluates complete
proposals in a single substantive review addressing both scientific and ethical
aspects of the proposal, following the general outline in Appendix B to this
guideline.
EPA will provide its completed review to the submitter. EPA's review may call
for revision of the proposal before it is reviewed by the HSRB. If EPA
determines that the proposal is ready for HSRB review, it will be scheduled for
HSRB review at the earliest opportunity. The submitter may respond to EPA's
review in writing to the HSRB docket, and may also comment orally at the HSRB
meeting.
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(iii) HSRB Review. As provided in 40 CFR §26.1601 (d), "following initial
evaluation of the protocol by Agency staff, EPA shall submit the protocol and all
supporting materials, together with the staff evaluation, to the Human Studies
Review Board." The HSRB is an advisory committee, chartered under the
Federal Advisory Committee Act, charged with advising EPA on scientific and
ethical aspects of proposed and completed research with human subjects. The
HSRB meets several times a year in public session, with opportunities for public
participation. All materials they consider are kept in a public docket, except any
which are subject to a substantiated claim of business confidentiality. HSRB
recommendations are documented in a public report of each meeting, prepared as
soon as possible after the meeting—typically within 60-90 days. As soon as it is
available, EPA will provide a copy of the HSRB's report to submitters of research
proposals discussed by the HSRB. Additional information about the HSRB,
including agendas and reports from past meetings and schedules of future
meetings, can be found at the HSRB website: www.epa.gov/osa/hsrb.
(e) Changes to IRB-approved research before execution. After EPA and the HSRB have
reviewed and commented on proposed research, revisions to the protocol, consent form
or other materials may be needed before the research is executed. When IRB-approved
materials are revised prior to initiation of the research to address concerns raised in EPA
and/or HSRB reviews or for any other reason, those revised materials must be re-
submitted to the IRB for review and approval before the research is initiated. This
requirement applies to all changes to an IRB-approved protocol, to all changes to an IRB-
approved consent form, and to all changes to any other materials approved by an IRB.
(40 CFR §26.1108(a)(4)). Although an IRB has the discretion to conclude that some
changes do not require review by the full IRB, investigators and sponsors do not have this
discretion. EPA will treat as a breach of the regulations any planned changes to proposed
research which are implemented without prior approval by the IRB.
(f) Execution of repellent studies
(1) Execution of protocol. When EPA and HSRB reviews are completed and the
IRB has approved any revisions resulting from those reviews, the research can be
initiated. Subjects can be recruited and data can be collected as specified in the
protocol. Care should be taken to ensure and document that these events occur in
the proper sequence. Initiation of subject recruitment before obtaining IRB
approval of the final protocol will be treated as a breach of the regulations.
(2) Quality Assurance (QA) oversight. Repellent efficacy testing, whether
conducted in the field or in the laboratory, is subject to the Good Laboratory
Practices regulations at 40 CFR part 160. These rules require that each testing
facility include an independent QA unit to monitor execution of each protocol and
document its conduct in accordance with the GLP regulations. (40 CFR §160.35)
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(3) Deviations from protocol. In executing even the best designed and most
comprehensive protocols, unanticipated deviations from the protocol may occur.
All such deviations from the protocol should be promptly and fully reported to the
1KB, and both the deviations and their impact on the research should be discussed
in the study report submitted to EPA. (40 CFR §160.185) EPA makes a
distinction between planned changes to a protocol and deviations from a protocol.
Planned changes must be treated as amendments to the protocol, and must be
approved by the IRB before they are implemented. (40 CFR §26.1108(a)(4))
(4) Changes in IRB-approved research after initiation. Any amendment to the
protocol, consent form, or other materials approved by the IRB must be submitted
to the IRB for review, and must be approved by the IRB before it is implemented,
unless immediate implementation is required to avert an imminent hazard to
subjects. (40CFR§26.1108(a)(4))
(5) Reports of adverse events. Any adverse events affecting the subjects of a
repellent efficacy test must be promptly reported to both the overseeing IRB and
to EPA, including adverse events not established to be related to product exposure
or study participation. (40 CFR §26.1108(b)(l))
(g) Reporting of completed repellent studies
(1) Study report. In addition to the standard elements required by the GLP
regulations at 40 CFR §160.185 and the appendices described below, the primary
report of the study should include these elements, as applicable.
(i) Study identification: Title, identifying study number(s), sponsor, study
director, investigators, name and location of the testing facility, and dates
of the study.
(ii) Approved or intended label directions for use.
(iii) Recruiting and enrollment statistics, including the following:
• Demographic characteristics of the pool from which subjects were
recruited and of the subject sample selected;
• Numbers of candidates contacted, interviewed, and screened; and
• Numbers of subjects enrolled, withdrawn voluntarily, withdrawn
involuntarily, and completing the research.
(iv) Test arthropods: Genus, species, subspecies or strain (if information is
available) of target pests; and pest pressure in each cage or field site. For
laboratory studies, the source of test arthropods, methods used to establish
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and maintain their disease-free status, rearing techniques and conditions,
developmental stage, age and sex of target arthropods and methods used to
sex them; preparation of arthropods before testing, and density per cage or
container.
(v) Test conditions: For field studies, the location and type of habitat, species
distribution, climate and environmental conditions. For laboratory studies,
a complete description of cages or containers, temperature, relative
humidity, ambient light, and air flow.
(vi) Test procedures, including but not limited to:
• Preparation of subjects: Training, demonstrating attractiveness,
clothing and protective equipment worn, calculation of skin area
and individual dose, washing, rinsing, and drying of limbs;
• Allocation of subjects to treatments; number of treatments per
subject; blinding key, if used;
• Method of dose determination;
• Method of application of repellent to each subj ect and individual
dose applied;
• Time of application of repellent to each subj ect;
• Time of start and end of exposure of each subj ect.
(vii) Complete accounting for all events during the testing period.
(viii) Results for each subject.
(ix) Reports of all statistical analyses
(x) Conclusions
(xi) Discussion
(2) Appendices.
(i) Complete protocol as approved by the IRB.
(ii) Consent form and any additional recruiting materials as approved by the
IRB.
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(iii) All amendments to the protocol, consent form, or other materials as
approved by the 1KB.
(iv) Reports of all deviations from the protocol and assessment of their impact
on the integrity of the research.
(v) Raw data and data collection sheets.
(vi) Documentation of ethical conduct of the research, as required by 40 CFR
§26.1303, including all correspondence with IRB not previously submitted
to EPA. The requirements of this section of the rule are summarized in
Appendix C to this guideline.
(h) Retention of Records. The following record-keeping requirements apply to some or all
records of research covered by this guideline.
(1) The record-keeping requirements in 40 CFR §26.1115 apply to Institutional
Review Boards (IRBs) that review human research conducted under EPA's
Human Research Rule.
(2) The record-keeping requirements of 40 CFR §169.2(j) apply to investigators who
conduct pesticide research with humans subject to FIFRA §12(a)(2)(P).
(3) The record-keeping requirements of 40 CFR §169.2(k) apply to any person who
submits the results of research to EPA in support of a petition for a tolerance or
tolerance exemption or in support of registration or an application for registration.
(4) The record-keeping requirements of 40 CFR §160.190 and §160.195 apply to
records of any study conducted under the Good Laboratory Practices rule.
(i) Specific guidance for dose-determination studies. All subjects participating in
repellency testing should be treated with the test material at a standard dose rate,
expressed by weight of repellent per unit area, mg/cm2 of treated skin, or volumetrically
as ml/cm2 of treated skin. Recommended methods for choosing the standard dose depend
in part on the active ingredient(s) and formulation(s) of the test material. For testing of
lotion formulations containing DEET (N,N-diethyl-meta-toluamide) eet, a standard dose
rate of 1 g per 600cm2 (equivalent to 1.67 mg per cm2) has been used in many tests of
repellency. Repellents in lotion form containing DEET may be tested for repellency at
this standard dose rate, or at a "typical consumer dose" rate determined empirically
through a dose-determination study. For other ingredients and formulations no
comparable standard dose has been identified, and it is recommended that a "typical
consumer dose" be estimated through empirical dose-determination testing as described
below. The rationale for the dose rate actually used in repellent testing should be
documented in all study reports.
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(1) Test material. The formulated product should be used as it is or will be
marketed, in the same type of container and bearing the same directions for use.
(2) Subjects. Subjects used for dose determination should be recruited and informed
as described above, and must provide written informed consent for their
participation. The same or very similar eligibility criteria would apply as for
repellency testing, although subjects participating only in dose-determination
testing would not be exposed to any insects or to any of the risks associated with
exposure to insects. The number of subjects used for dose determination should
be justified statistically. The sample should include adults of both sexes and
varied race, and should be as representative as possible of the repellent user
population, consistent with ethical and feasibility constraints.
(3) Measuring subject's skin area. The surface area of subject's limb in cm2 can be
estimated by measuring the circumference of the forearm in centimeters at the
wrist and elbow, or of the leg at the ankle and knee, and in either case at one or
two equally spaced intermediate points; then multiplying the average
circumference by the length of the limb in centimeters from the wrist to elbow or
from the ankle to the knee.
(4) Methods. Each subject should be instructed to apply the test repellent to his or
her own limbs—to forearms or lower legs, or both—as they normally would
apply a repellent to achieve complete coverage. Once the quantity applied has
been measured and recorded, the applied repellent should be washed completely
off the limb. This process should be repeated at least 3 times for each limb
treated by each subject.
The applied quantity of a lotion or towelette formulation can be determined by
weighing the container or towelette before and after use. Because less than all the
spray released from a pump-spray or aerosol container will reach the target limb,
a different method is needed to estimate the applied quantity of a spray
formulation. The target limb should be wrapped in gauze "bracelets" of known
area so that the bracelets cover only part of the skin. So long as the area of the
bracelets is known, the quantity of a spray formulation applied to the skin per cm2
can be estimated by weighing the bracelets before and after application. Skin
should be left exposed between bracelets to help subjects determine when they
achieve complete coverage. Dose determination with spray formulations should
be conducted out-of-doors.
(5) Calculating standard dose for use in repellency trials. The mean dose applied
by each subject to each limb and the grand mean across all subjects of mean doses
applied to forearms and to lower legs should be calculated. The specific gravity
of the test material should be used to convert the dose expressed by weight as mg
per cm2 to a volumetric dose expressed as ml per cm2; this calculation should be
included in the study report. The volumetric dose should be used as the standard
dose for repellency testing, scaling it to the treated surface area of each subject's
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limb or limbs. The protocol should include the specific gravity of all test
materials and should specify this calculation, and should specify doses both by
weight and by volume.
(j) Specific guidance for laboratory studies of mosquito or biting fly repellency.
(1) Species. Mosquito tests should be conducted with Aedes aegypti, an Anopheles
species such as An. albimanus, or a Culex species, preferably Cx.
quinquefasciatus. Stable fly tests should be conducted with Stomoxys calcitrans.
The source of test insects should be reported, and if possible, they should be
identified by subspecies or strain.
(2) Stage, age, and sex. Mosquito testing should be conducted with adult females 5-
10 days old; methods of sexing should be reported. Stable flies should be male or
female adults 3-10 days old. The age range of test insects should be reported.
(3) Rearing techniques. Mosquito and biting fly larvae should be reared under
optimal conditions for the species—typically at 27° ± 3°C, relative humidity 80%
± 10%, and photoperiod 16:8 hours (light:dark). Alternative rearing methods for
mosquitoes are discussed by Gerberg et al. (1998). Use of alternative rearing
techniques for biting flies and/or mosquitoes should be fully documented and
justified.
(4) Preparation of insects. Test mosquitoes should be fed 10% sucrose and no
blood meal before use in a test, and starved for 12 to 24 hours immediately before
the test. Test stable flies may be fed dry sugar cubes with a separate water source;
they should also receive no blood meal before use in a test, and should be starved
for 24 hours immediately before the test. Test insects should be established to be
free of disease; the source of insects and the methods used to ensure they are
disease-free should be reported.
(5) Disposition of insects. Test insects should be used only once and destroyed
immediately after the trial.
(6) Test cages. Cage size may vary, but is typically 2' x 2' x 2' (8 cubic feet in
volume, roughly equivalent to 232,000 cm3), square or rectangular in plan, with
one or more sleeved openings for the subject's arms. A mirror on the bottom of
the cage helps observers see insects landing on the underside of subjects' arms.
Larger cages may be used for two subjects at a time, so long as they are treated
with the same repellent. Cage dimensions and design should be reported in detail.
(7) Insect density. There should be at least 200 mosquitoes in each 2'x2'x2' cage
(equivalent to one for each 1,160 cm3) or at least 50 stable flies per cage (one for
each 4,640 cm3.) After each exposure period all stable flies should be removed by
vacuuming from each cage, and 50 fresh stable flies should be added to each cage.
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(8) Testing conditions. Ambient temperature during the test should be maintained at
21° ± 8°C, and relative humidity at 55-85%. Ambient light appropriate to the
preferred feeding time for the test species should be provided—e.g., more light
for day feeders like Aedes aegypti, and less for night feeders like Anopheles spp.
and Culex spp. When testing in subdued light, care should be taken not to
compromise the investigators' ability to observe insect activity. Investigators and
subjects should avoid exhaling into the test cage; introduction of CC>2 could bias
insects towards biting.
(9) Treated area size and preparation. The recommended treatment area is the
subject's forearm (wrist to elbow). This area should be washed with unscented
soap, rinsed with a solution of ethanol or isopropyl alcohol in water, and dried
with a clean towel. The surface area of each test subject's treated forearm (in
cm2) and the volumetric dose administered to each subject (in ml) should be
reported. Skin adjacent to the treated area should be covered with a light-colored
material that test insects cannot bite through. Hands should be covered with
impenetrable (e.g., latex) gloves.
(10) Untreated controls. One or two untreated control subjects should verify
continued landing pressure in tests of CPT. Results for these untreated subjects
should not be compared to those for treated subjects; multiple untreated controls
should be used as a standard of comparison only in studies of relative
protection/percent repellency. To minimize potential interference, untreated
subjects should not be treated on their other forearm. Forearms of untreated
subjects should be washed, rinsed, and dried exactly like treated forearms.
(11) Positive controls. A positive control group treated at a rate of 1 ml per 600 cm2
with 20% DEBT in ethanol is recommended to calibrate the test system.
(12) Establishing subject attractiveness to test insects. Before the test, subjects
should expose their untreated forearms to the target insects in a test cage to
establish their attractiveness. Five mosquito landings in one minute or less or 2
stable fly landings in two minutes or less are sufficient to establish attractiveness.
(13) Establishing and confirming landing pressure. Before exposing any treated
subjects during each exposure period, an untreated subject's forearm should be
inserted into each cage to confirm landing pressure. Landing insects should be
removed from the subject's arm, by shaking it or by other means, before they
have time to bite. The untreated subject should remove his or her forearm from
the cage as soon as it has received the requisite number of landings. If at any time
fewer than 5 mosquitoes land on the untreated control forearm within one minute,
all mosquitoes should be removed from all cages in the study and fresh insects
should be added to each cage. If at any time fewer than 2 stable flies land on the
untreated control forearm within two minutes, all stable flies should be removed
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from that cage and 50 fresh flies should be added. The aggressiveness of the fresh
insects should be confirmed before repellency testing continues.
(11) Exposure period. Approximately 30 minutes after treatment with the repellent,
test subjects should insert their treated forearms into the cage for the first time.
Exposures of pre-defined duration should be repeated at regular intervals—e.g., 5-
minute exposures at 30-minute intervals—until efficacy failure for each subject or
the end of the study, whichever occurs first. Subjects should avoid rubbing their
arms when inserting them into or removing them from the cage and between
exposure periods.
(k) Specific Guidance for field studies of mosquito or biting fly repellency.
(1) Pre-test subject preparation. Subjects in field studies of repellency should be
trained in the laboratory to identify landings, probes, and bites of the target
species and to use aspirators to collect landing insects before they have time to
bite.
(2) Choice of field testing sites. Field tests for mosquito repellency should be
conducted in at least two distinct habitats (e.g., forest, grassland, salt marsh,
wetland, beach, barns, or urban environments) where the predominant mosquito
species differ. So long as the environments and species distribution differ, it is
not important for test sites to be widely separated geographically. Repellency
against black flies, stable flies, or other biting flies typically occurring in only one
habitat may be tested in only one habitat.
Potential sites for testing mosquito repellency should be monitored at least weekly
for a month before testing is scheduled. To minimize risks to subjects, field
testing should not be conducted where West Nile virus (WNV) or other mosquito-
vectored diseases have been detected within the previous two weeks. Because
biting flies are not known to vector human diseases in the USA, no comparable
site monitoring is required for studies of repellency to biting flies.
(3) Species identification. Mosquito tests should be conducted where more than one
species is present. If tests are conducted outside the U.S., the relevance of the
study for U.S. regulatory purposes should be justified in the study report.
Landing insects should be aspirated or trapped before and during the test, and
labeled with the time of collection. After the field study, collected insects should
be identified by genus and species, and if possible, by subspecies or strain. The
number in each taxon collected in each time period should be reported.
After identification, mosquitoes should be subjected to serological or other
analysis to determine the presence or absence of WNV or other disease
organisms. The results of these analyses should be reported to subjects and
included in the study report.
27
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(4) Untreated controls. Untreated control subjects are necessary in all study
designs. To minimize the potential for interference, untreated controls should be
subjects who are not treated with a repellent on any limb.
When the objective is to estimate CPT, two untreated control subjects are
sufficient. They should expose an untreated arm or leg briefly at regular intervals
during the test to confirm continued acceptable landing pressure. Results for the
untreated controls should not be compared to results for treated subjects. When
the objective is to evaluate relative protection, more untreated control subjects are
needed to provide a standard of statistical comparison.
Untreated limbs used as controls should be washed, rinsed, and dried exactly like
treated limbs.
(5) Positive controls. A positive control group treated at a rate of 1 ml per 600 cm2
with 20% deet in ethanol is recommended to calibrate the test system.
(6) Minimum landing pressure to initiate or continue test. Landing pressure
should be measured before treatment and intermittently throughout the course of
the test by untreated control subjects. Testing should not be conducted or
continued unless landing pressure of the target species is at least one mosquito
landing within one minute, or at least one stable fly, black fly, ceratopogonid or
tabanid landing within five minutes. Insects landing on untreated subjects should
be collected for later identification, and labeled with the time of collection.
(7) Treated area size and preparation. The recommended treatment area is the
subject's forearm (wrist to elbow) or lower leg (ankle to knee), depending on the
feeding behavior of the predominant species at the selected test site. A smaller
treated area on the appropriate limb of at least 250 cm2 is also acceptable. The
treated area should be washed with unscented soap, rinsed with a solution of
ethanol or isopropyl alcohol in water, and dried with a clean towel. The surface
area treated for each test subject limb and the specific dose administered (ml) to
each subject should be recorded. With the exception of the treated area, subjects'
heads, trunks, and limbs should all be covered with light-colored material through
which insects cannot bite.
(7) Exposure period. Exposure of treated subjects may be intermittent—for
example, for 1 or 2 minutes at 15-minute intervals, or for 5 minutes at 30-minute
intervals—to reduce risks to subjects and allow them to rest between exposures.
When testing a repellent with a long period of effectiveness, it is critical to the
reliability of the data to minimize "right-censoring" of the data—i.e., the number
of subjects who do not experience a failure of efficacy before the end of the test.
Reliable results may be obtained for extended periods of protection by treating
subjects up to several hours before the first field exposure, with subsequent
28
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exposures timed to coincide with periods of target insect activity. This approach
reduces prolonged exposure of subjects to insects in the field, helps to reduce
early withdrawal of subjects attributable to excessively long trials, and increases
the likelihood that most or all subjects will experience a failure of efficacy.
Another acceptable design for assessing long-term repellency involves treating
different subjects at different times (e.g., 2, 4, 6, 8, or 10 hours before exposure)
and then exposing all subjects at the same time when target insects are active.
(8) Minimizing variation. Many factors contributing to variability in repellent field
studies cannot be controlled, but minimizing variation when possible and
designing studies with insect feeding patterns in mind can make a study design
more efficient. Standardization is possible for such factors as time of testing,
allocation to treatment, subject attractiveness to target insects, etc. Other factors
which may be managed to minimize variation include targeting a predominant
species in its habitat, synchronizing exposure periods with peak feeding activity
of target insects, and treating limbs consistent with the preference of the target
species to feed on legs, arms, or both.
(9) Environmental conditions. The time of day at which subjects are treated and at
which exposure to target insects begins and ends should be recorded and reported.
Weather conditions during testing (including temperature, relative humidity,
cloud cover, precipitation, light intensity, and wind speed) should be monitored
periodically throughout the study and reported. Testing should be not be
conducted or continued if wind speed exceeds 16 kph/10 mph.
(10) Test subject placement and behavior. Subjects may work in pairs to assist each
other to identify landings and collect insects. Because clustering of subjects may
confound results, at least 3 m/10 ft should be maintained between pairs. Subjects
should avoid strenuous exercise and sweating, since these could affect test results.
(11) Data collection and reporting. Under the supervision of the investigator, an
associate or another subject should record the number and timing of each landing
or bite during each exposure period for each subject. If possible, all landing
insects should be collected for identification and labeled with the time of
collection.
(1) Specific guidance for laboratory studies of tick and chigger repellency. Because
reliable field tests have not been developed, EPA recommends testing for repellency
against ticks and chiggers in the laboratory. If field tests are conducted, proposed label
directions for re-application frequency should reflect field study results.
(1) Summary of recommended method. Each subject should place the fingers of
one hand on a flat surface, with the elbow above the wrist and the forearm held at
an angle of 30° or more to the surface. With a suitable instrument (such as an
29
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artist's paintbrush, forceps, or a cotton swab) the investigator should place a tick
or chigger, one at a time, on the subject's wrist, at a release point marked 3 cm
below the boundary of the treated area of the forearm. The tick or chigger should
be oriented gently (e.g., with paint brush, forceps, or cotton swab) toward the
treated area. After its first movement up the arm toward the margin of the treated
area, each tick or chigger should be allowed 3 minutes to move across the
boundary onto the treated area. A tick or chigger that crosses at least 3 cm into
the treated area (toward the elbow) is reported as 'not repelled'. One that does not
cross into the treated area, or that crawls into the treated area but immediately
turns back or falls off, is reported as 'repelled.' Fresh ticks or chiggers are
exposed to the treated area one at a time, at regular intervals for the duration of
the test.
While the general definition of a "crossing" is constant, the details of the method
of scoring a crossing must be operationalized in each protocol in a way
appropriate to the species and life stage of ticks to be tested. A release point 3 cm
distant from the treated area and scoring of a crossing when a tick moves at least 3
cm into the treated area have worked effectively in tests with nymphal
Dermicentor variabilis and Ixodes scapularis, and are provided here as examples.
(2) Species. Tick tests should be conducted using laboratory colonies of the tick
species the label claims to repel. Common tick species in the United States
include the blacklegged tick (deer tick, Ixodes scapularis), western blacklegged
tick (deer tick, Ixodespacificus\ lone star tick (Amblyomma americanum\
American dog tick (Dermacentor variabilis), and relapsing fever tick (softbacked
tick, Ornithodoros turicatd). Chigger tests should be conducted using laboratory
colonies of chiggers in the Trombiculidae family; Eutrombicula splendens; or E.
cinnabarrs. Test arthropods should be identified by genus and species, and if
possible by subspecies or strain. It is permissible to test two species concurrently,
alternating so that each subject qualifies and tests a tick or chigger from each
species within each exposure period.
(3) Stage, age, and sex. When testing with blacklegged (deer tick), lone star, or soft-
backed ticks, either adult or nymphal life stages are appropriate for testing. Only
the adult American dog tick is recommended, since nymphs of this species do not
feed on humans. Tests with chiggers should use immature chiggers. The age or
age range of all test arthropods should be reported.
(4) Rearing techniques. Ticks and chiggers should be reared at 25° ± 3°C, at high
relative humidity (> 90%), and photoperiod of approximately 16:8 hours
(lightdark). Any alternative rearing techniques should be fully described and
justified.
(5) Preparation of test arthropods. Ticks and chiggers should be disease-free. The
source of test animals and the methods used to ensure they are disease-free should
be reported.
30
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(6) Disposition of test arthropods. Each tick or chigger should be used only once
and should be destroyed immediately after use.
(7) Number of ticks or chiggers. Ticks or chiggers are tested for questing behavior
and for repellency one at a time, according to the study design. Some studies
using a single species of tick test 5 ticks in each 30-minute exposure period; other
studies test one tick of each of two species in each 15-minute exposure period.
Either pattern is acceptable.
(8) Testing conditions. Temperature should be maintained during the test at 20° to
25°C, with relative humidity at > 35% and indirect ambient light 50 to 80%. The
lights should be kept on.
(9) Subject preparation. Before treatment, both forearms of each subject are
washed with unscented detergent and carefully rinsed and dried. One forearm
from wrist to elbow is treated with the test material, and two lines are drawn on
the subject's wrist: a 'boundary line' at the edge of the treated area, and another
line—the 'release line'—3 cm distant from the boundary line, outside the treated
area toward the fingers. Similar lines are drawn in the same positions on the wrist
of the subject's other, untreated arm—one line at the wrist, and another 3 cm
away toward the fingers.
(10) Untreated control. The untreated forearm of each treated subject is used to
screen ticks or chiggers for questing behavior; only actively questing ticks or
chiggers should be selected for repellency testing. With an appropriate
instrument, such as an artist's paint brush, forceps, or a cotton swab, each tick or
chigger should be picked up carefully to prevent damaging its body or forelegs
and placed on the release line on the wrist of the subject's untreated arm. A tick
or chigger that moves steadily from the release line across the boundary line and
upward along the subject's untreated forearm is actively questing, and appropriate
for use in repellency testing. A tick or chigger failing this test should be
immediately destroyed.
(11) Positive controls. A positive control group treated at a rate of 1 ml per 600 cm2
with 20% deet in ethanol is recommended to calibrate the test system.
(12) Test procedure. With the treated forearm held upright above the table, one fresh
tick or chigger at a time, immediately after demonstrating active questing on the
subject's untreated arm, is placed at the release line on the treated arm. A
"crossing" is recorded if the test organism crosses the boundary line at least 3 cm
into the treated area within 3 minutes, and remains in the treated area for at least
one minute.
(13) Exposure period. At intervals of 15 to 30 minutes according to the study design,
fresh ticks or chiggers are placed one at a time at the release point on the
31
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untreated arm and tested for active questing. Qualified ticks or chiggers are then
placed at the release point on the treated arm and exposed to the treated area. The
cycle continues until efficacy failure or the end of the study, whichever occurs
first.
(14) Data collection and reporting. Recording of crossings and repelled test ticks or
chiggers arthropods during the exposure period should be supervised by
investigators. The investigator, an associate, or another subject should record the
number and timing of all events occurring during in each exposure period for each
subject.
(m) References. The following publications were consulted in developing this guideline.
(1) American Society for Testing and Materials. (2006) E 939-94 Standard Test
Method of Field Testing Topical Applications of Compounds as Repellents for
Medically Important and Pest Arthropods (Including Insects, Ticks and Mites): I
Mosquitoes.
(2) American Society for Testing and Materials. (2003) E 1488-02 Standard Guide
for Statistical procedures to use in Developing and Applying test methods.
(3) Barnard, D.R.; Ulrich, R.B.; Xue, R.; and Debboun, M. (2007) Chapter 5:
Standard methods for testing mosquito repellents. In Insect Repellents: Principles,
Methods and Uses. Debboun, Frances, and Strickman, eds. CRC Press. (495 p.)
(4) Bernard, D. R., Bernier, U. R., Posey, K. H. and Xue, R-D (2002). Repellency of
IR 3535, KBR 3023, para-menthane-3,8,-diol, and deet to Black Salt Marsh
Mosquitoes (Diptera:Culicidea) in Everglades National Park. J. Med.
Entomology39(6): 895-899
(5) Barnard, D.R. (1998) Mediation of Deet repellence in mosquitoes (Diptera:
Culicidae) by species, age, and parity. J. Med. Entomol. 35(3): 340-343.
(6) Barnard, D.R.; Posey, K.H.; Smith, D.; and Shreck, C.E. (1998) Mosquito
density, biting rate and cage size effects on repellent tests. Med. Vet. Entomol.
12:39-45.
(7) Carroll, S. P. (2007) Chapter 12: Evaluation of topical insect repellents and
factors that affect their performance. In Insect Repellents: Principles, Methods
and Uses. Debboun, Frances, and Strickman, eds. CRC Press. (495 p.)
(8) Frances, S.P. (1994) Response of a chigger, Eutrombicula hirsti (Acari:
Trombiculidae) to repellent and toxicant compounds in the laboratory. J. Med.
Entomol. 31(4): 628-630.
32
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(9) Frances, S.P.; Eikarat, N.; Sripongsai, B.; and Eamsila, C. (1993) Response of
Anopheles dims and Aedes albopictus to repellents in the laboratory. J. Am. Mos.
Con. Assoc. 9(4): 474-476.
(10) Gerberg, E. J. ; Barnard, D.R.; and Ward, R.A. (1998) Manual for Mosquito
Rearing and Experimental Techniques. AMCA Bull. 5.
(11) Govere, J.M.; and Durrheim, D. N. (2007) Chapter 8: Techniques for evaluating
repellents. In Insect Repellents: Principles, Methods and Uses. Debboun, Frances,
and Strickman, eds. CRC Press. (495 p.)
(12) Huntsberger, D. and Billingsley, P. (1981) Elements of Statistical Inference. 5th
edition. Allyn and Bacon. Inc., Boston.
(13) Klun, J.A. and Debboun, M. (2000) A new module for quantitative evaluation of
repellent efficacy using human subjects. J. Med. Entomol. 37(1): 177-181.
(14) Schofield, S.; Tepper, M.; and Gadawski, R. (2007) Field evaluation against
mosquitoes of regular and polymer-based deet formulations in Manitoba, Canada,
with comment on methodological issues. J. Med. Entomol. 44: 457-62.
(15) Smith, C.N. (1955) Insect repellents. Quarterly Report, Entomological Research.
Entomology Research Branch, U.S. Department of Agriculture. 8 p.
(16) Verwey, R.E. (1996) Laboratory method for testing insect repellents on human
test subjects against chiggers in the laboratory. Unpublished document prepared
by S.C. Johnson & Sons, Inc., Racine, WI. 3 p.
(17) WHO (2009) Guidelines for Efficacy Testing of Mosquito repellents for Human
Skin: CDS/NTD/WHOPES/2009.4.
(18) WHO (1996) Report of the WHO Informal Consultation on the Evaluation and
Testing of Insecticides: CTD/WHOPES/1C/96.1.
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Appendix A: Checklist of Elements Required by 40 CFR §26.1125
40 CFR 26.1125 Prior submission of proposed human research for EPA review
Any person or institution who intends to conduct or sponsor human research covered by §26.1101 (a) shall, after receiving
approval from all appropriate IRBs, submit to EPA prior to initiating such research all information relevant to the proposed
research specified by §26.1115(a), and the following additional information, to the extent not already included:
Requirement Y/N Comments/Page Refs
All information relevant to the proposed research specified by § 26.1 1 1 5(a)
The following Information, to the
extent not already included:
(1) Copies of
• all research proposals reviewed by the IRB,
• scientific evaluations, if any, that accompanied the proposals
reviewed by the IRB,
• approved sample consent documents,
• progress reports submitted by investigators, and reports of injuries
to subjects.
(2) Minutes of IRB meetings ... in sufficient detail to show
• attendance at the meetings;
• actions taken by the IRB;
• the vote on these actions including the number of members voting
for, against, and abstaining;
• the basis for requiring changes in or disapproving research;
• a written summary of the discussion of controverted issues and
their resolution.
(3) Records of continuing review activities.
(4) Copies of all correspondence between the IRB and the investigators.
(5)
• A list of IRB members identified by name; earned degrees;
representative capacity; indications of experience such as board
certifications, licenses, etc., sufficient to describe each member's
chief anticipated contributions to IRB deliberations;
• any employment or other relationship between each member and
the institution, for example, full-time employee, a member of
governing panel or board, stockholder, paid or unpaid consultant.
(6) Written procedures for the IRB in the same detail as described in
§26.1 108(a) and §26.1 108(b).
(7) Statements of significant new findings provided to subjects, as required
by§26.1116(b)(5).
(1) The potential risks to human subjects
% (2) The measures proposed to minimize risks to the human
^ § subjects;
LO" '(/5 (3) The nature and magnitude of all expected benefits of such
•<- = research, and to whom they would accrue
«« .£ (4) Alternative means of obtaining information comparable to what
co would be collected through the proposed research; and
(5) The balance of risks and benefits of the proposed research.
§1125(b): All information for subjects and written informed consent
agreements as originally provided to the IRB, and as approved by the IRB.
§1 125(c): Information about how subjects will be recruited, including any
advertisements proposed to be used.
§1 125(d): A description of the circumstances and methods proposed for
presenting information to potential human subjects for the purpose of
obtaining their informed consent.
§1125(e): All correspondence between the IRB and the investigators or
sponsors.
§1 125(f): Official notification to the sponsor or investigator . . . that
research involving human subjects has been reviewed and approved by
an IRB.
34
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Appendix B: Framework for Science and Ethics Reviews of Proposed Human Research
Questions to be Addressed in EPA Protocol Review
Protocol Identification
(a) Title
(b) Date
(c) Principal Investigator and any sub-investigators
(d) Participating Laboratories
(e) Sponsor
(f) Reviewing 1KB
1. Societal Value of Proposed Research
(a) What is the stated purpose of the proposed research?
(b) What research question does it address? Why is this question important? Would the
research fill an important gap in understanding?
(c) How would the study be used by EPA?
(d) Could the research question be answered with existing data? If so, how?
(e) Could the question be answered without newly exposing human subjects? If so, how?
2. Study Design
(a) What is the scientific objective of the study? If there is an explicit hypothesis, what is it?
(b) Can the study as proposed achieve that objective or test this hypothesis?
2.1 Statistical Design
(a) What is the rationale for the choice of sample size?
(b) What negative and positive controls are proposed? Are proposed controls appropriate
for the study design and statistical analysis plan?
(c) How is the study blinded?
(d) What is the plan for allocating individuals to treatment or control groups?
(e) Can the data be statistically analyzed?
(f) What is the plan for statistical analysis of the data?
(g) Are proposed statistical methods appropriate to answer the research question?
(h) Does the proposed design have adequate statistical power to definitively answer the
research question?
2.2 How and to what will human subjects be exposed?
(a) To what will subjects be exposed?
(b) What is the rationale for the choice of test material and formulation?
(c) What is the rationale for the choice of dose/exposure levels and the staging of dose
administration?
(d) What duration of exposure is proposed?
35
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Appendix B: Framework for Science and Ethics Reviews of Proposed Human Research
2.3 Endpoints and Measures
(a) What endpoints will be measured? Are they appropriate to the question(s) being
asked?
(b) What steps are proposed to ensure measurements are accurate and reliable?
(c) What QA methods are proposed?
(d) How will uncertainty be addressed? Will reported point values be accompanied by
measures of uncertainty?
3. Subject Selection
3.1 Representativeness of Sample
(a) What is the population of concern? How was it identified?
(b) From what populations will subjects be recruited?
(c) Are expected participants representative of the population of concern? If not, why
not?
(d) Can the findings from the proposed study be generalized beyond the study sample?
3.2 Equitable Selection of Subjects
(a) What are the inclusion/exclusion criteria? Are they complete and appropriate?
(b) What, if any, is the relationship between the investigator and the subjects?
(c) If any potential subjects are from a vulnerable population, what is the justification for
including them?
(d) What process is proposed for recruiting and informing potential subjects?
(e) If any subjects are potentially subject to coercion or undue influence, what specific
safeguards are proposed to protect their rights and welfare?
3.3 Remuneration of Subjects
(a) What remuneration, if any, is proposed for the subjects?
(b) Is proposed remuneration so high as to be an undue inducement?
(c) Is proposed remuneration so low that it will only be attractive to economically
disadvantaged subjects?
(d) How and when would subjects be paid?
4. Risks to Subjects
4.1 Risk characterization
(a) Have all appropriate prerequisite studies been performed? What do they show about
the hazards of the test materials?
(b) What is the nature of the risks to subjects of the proposed research?
(c) What is the probability of each risk associated with the research? How was this
probability estimated?
36
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Appendix B: Framework for Science and Ethics Reviews of Proposed Human Research
4.2 Risk minimization
(a) What specific steps are proposed to minimize risks to subjects?
(b) How do proposed dose/exposure levels compare to established NOELs/NOAELs for
the test materials?
(c) What stopping rules are proposed in the protocol?
(d) How does the protocol provide for medical management of potential illness or injury
to subjects?
(e) How does the protocol provide for safety monitoring?
(f) How does the protocol provide for post-exposure monitoring or follow-up? Is it of
long enough duration to discover adverse events which might occur?
(g) How and by whom will medical care for research-related injuries to subjects be paid
for?
5. Benefits
(a) What benefits of the proposed research, if any, would accrue to individual subjects?
(b) What benefits to society are anticipated from the information likely to be gained through
the research?
(c) How would societal benefits be distributed? Who would benefit from the proposed
research?
(d) What is the likelihood that each identified societal benefits would be realized?
6. Risk/Benefit Balance
(a) How do the risks to subjects weigh against the anticipated benefits of the research, to
subjects or to society?
7. Independent Ethics Review
(a) What Institutional Review Board (1KB) reviewed the proposed research?
(b) Is this 1KB independent of the investigators and sponsors of the research?
(c) Is this 1KB registered with CHRP?
(d) Is this 1KB accredited? If so, by whom?
(e) Does this 1KB hold a Federal-Wide Assurance from CHRP?
(f) Are complete records of the 1KB review as required by 40 CFR §26.1125 provided?
(g) What standard(s) of ethical conduct would govern the work?
8. Informed Consent
(a) Will informed consent be obtained from each prospective subject?
(b) Will informed consent be appropriately documented, consistent with the requirements of
40 CFR §26.1117?
(c) Do the informed consent materials meet the requirements of 40 CFR §26.1116, including
adequate characterization of the risks and discomforts to subjects from participation in
the research, the potential benefits to the subject or others, and the right to withdraw from
the research?
37
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Appendix B: Framework for Science and Ethics Reviews of Proposed Human Research
(d) What is the literacy rate in English or other languages among the intended research
subjects?
(e) What measures are proposed to overcome language differences, if any, between
investigators and subjects?
(f) What measures are proposed to ensure subject comprehension of risks and discomforts?
(g) What specific procedure will be followed to inform prospective subjects and to seek and
obtain their consent?
(h) What measures are proposed to ensure fully voluntary participation and to avoid coercion
or undue influence?
9. Respect for Subjects
(a) How will information about prospective and enrolled subjects be managed to ensure their
privacy?
(b) How will subjects be informed of their freedom to withdraw from the research at any
time without penalty?
(c) How will subjects who decline to participate or who withdraw from the research be dealt
with?
38
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Appendix C: Checklist of Elements Required by 40 CFR §26.1303
§ 26.1303 Submission of Completed Human Research for EPA Review
Any person who submits to EPA data derived from human research covered by this subpart shall provide at the time of submission
information concerning the ethical conduct of such research. To the extent available to the submitter and not previously provided to
EPA, such information should include:
Requirement
evant to the research specified by
d and maintained by an IRB
CD CD
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8 ^
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I of the records relevant to
dentified in §26.11 25(a)-(f)
(b) Copies of a
the information
§1115(a)(1): Copies of
• all research proposals reviewed,
• scientific evaluations, if any, that accompany the proposals,
• approved sample consent documents,
• progress reports submitted by investigators, and reports of injuries to
subjects.
§1 1 15(a)(2): Minutes of IRB meetings which shall be in sufficient detail to show
• attendance at the meetings;
• actions taken by the IRB;
• the vote on these actions including the number of
• members voting for, against, and abstaining;
• the basis for requiring changes in or disapproving research;
• a written summary of the discussion of controverted issues and their
resolution.
§1115(a)(3): Records of continuing review activities.
§1115(a)(4): Copies of all correspondence between the IRB and the investigators.
§1115(a)(5):
• A list of IRB members identified by name; earned degrees; representative
capacity; indications of experience such as board certifications, licenses,
etc., sufficient to describe each member's chief anticipated contributions
to IRB deliberations;
• any employment or other relationship between each member and the
institution, for example, full-time employee, a member of governing panel
or board, stockholder, paid or unpaid consultant.
§1115(a)(6): Written procedures for the IRB in the same detail as described in §
26.1108(a) and § 26.1 108(b).
§1 1 15(a)(7): Statements of significant new findings provided to subjects, as
required by § 26.11 16(b)(5).
^ (1 ) The potential risks to human subjects;
„ ° (2) The measures proposed to minimize risks to the human subjects;
•5- .2 (3): The nature and magnitude of all expected benefits of such research,
CM
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