United States Environmental Protection Agency Office of Chemical Safety and Pollution Prevention (7101) EPA 712- [insert number] [insert date, using “DRAFT” before the date, until finalized] 

# Product Perfomance Test Guidelines

OPPTS 810.3700 : Insect Repellents to be Applied to Human SkinNOTICE

This is one of a series of test guidelines established by the Office of Chemical Safety and Pollution Prevention (OCSPP), 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, et seq. ), and section 408 of the Federal Food, Drug and Cosmetic Act (FFDCA) (21 U.S.C. 346a).

OPPTS test guidelines serve as a compendium of accepted scientific methods for research intended to provide data to inform regulatory d ecisions 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 da ta 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. Th e Agency will assess such proposals on a case-by-case basis.

For additional information about OCSPP harmonized test guidelines and to access the guidelines electronically, please go to http://www.epa.gov/ocspp 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.

DRAFT DOCUMENT DISCLAIMER: This draft document is distributed solely for the purpose of external review. It has not been formally dissemi nated by the EPA and should not be construed to represent any Agency determination or policy. The information correction process under the Agency’s Information Quality Guid elines does not apply until this doc ument is formally disseminated by the EPA in its final form. This draft documen t should only be cited or quoted in the context of providing comments.OPPTS 810.3700: Insect Repellents to be Applied to Human Skin

(a) Introduction.

(1) Scope and Purpose. This guideline provides r ecommendations for the design and execution of studies to evaluate the performance of pesticide products intended to repel insects and other arth ropods in connection with the products’ registration under the Federal Insectic ide, Fungicide, and Rodenticide Act (FIFRA) (7 U.S.C. 136, et seq. ). This guidance applie s to products in any formulation—such as lotion, liquid, or spra y—intended to be a pplied directly to human skin. It does not apply to products applied to or impr egnated into clothing or fabric, or used to repe l insects from indoor or out door spaces. This guidance recommends appropriate study designs and methods for selecting subjects, statistical analysis, and reporting.

(2) General Considerations. Any protocol developed us ing this guidance must meet the requirements set forth in several st atutes and regulations , including, but not limited to, the Federal Insecticide, F ungicide, 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 s ubjects of research, 40 CF R part 26, subparts K through Q. Because these studies would s upport a FIFRA registration and involve intentional exposure of huma n subjects to the test re pellents, review of each protocol by an Institutiona l 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 ove rrule 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 regula tion, 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 a ny 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 me ntal health consequences which are reasonably foreseeable therefrom, and (i i) freely volunteer to participate in the test.” Regulations implementing this statutory provi sion and defining associated record-keeping requireme nts 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 guidelin e is covered by the requirements of EPA regulations for the protection of hu man subjects of research set out at

140 CFR part 26, subparts K, L, and M. Persons conducting and submitting topical repellent efficacy studies sh ould 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 s ubjects 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 c onnection with any action that may be performed by EPA under FIFR A 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 in tentional exposure of any human subject who is a pregnant woma n (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 approv al), or disappro ve all research activities covered by this subpart.”

(D) Required Pre-testing submissions to EPA. 40 CFR §26.1125 provides:

“Any person or inst itution who intends to conduct or sponsor human research covered by §26.1101( a) shall, af ter receiving approval from all appropriate Inst itutional Review Boards (IRBs), submit to EPA prior to initiating such research all information relevant to the proposed resear ch specified by §26.1115(a), and, to the extent not already included:

2(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 a nd benefits of the proposed research.

(b) All information for subjects and written informed consent agreements as originally pr ovided to the IRB, and as approved by the IRB.

(c) Information about how subj ects 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 th eir informed consent.

(e) All correspondence between th e IRB and the investigators
or sponsors.

(f) Official notification to the sponsor or investigator, in accordance with the requiremen ts of this subpart, that research involving human subjects has been reviewed and approved by an IRB.”

(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 reco rds relevant to the research specified by §26.1115(a) to be prepared and maintained by an IRB.

(b) Copies of all of the records relevant to the information identified in §26.1125(a) through (f).

3(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 though L of this part.”

(iii) Good Laboratory Practice Standards. Good Laboratory Practice Standards (GLP) as defined in 40 CF R part 160 apply to both laboratory and field studies of repellent efficacy. According to 40 CFR §160.17: “EPA may refuse to consider reli able for purposes of supporting an application for a research or marke ting permit any data from a study which was not conducted in accordance wi th this part.” 40 CFR §160.12(b) requires with any submitted research data “[a] statement describing in detail all differences between the pr actices used in the study and those required by this part.” Additiona lly, 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 a pplicable 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 repellen t efficacy testing, whether conducted in the laborat ory or in the field, and to all target species. Each of these six sections discusses one of th e 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.

4(g) Reporting of completed repellent studies. (h) Retention of records.

Sections (i) through (l) provide guidance specific to diffe rent 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 st udies of mosquito or biting fly repellency. (k) Specific guidance for field studies of mosquito or biting fly repellency. (l) Specific guidance for laboratory st udies 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 Scie nce 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 spec ial importance in understanding this guideline. They apply only in the context of this guideline and ar e 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 jump ing insect or other arthropod alighting on human skin w ithout probing or biting.

(ii) A probe is the act of penetrating huma n skin by the mouthparts of an insect or other arthropod without ingestion of blood.

(iii) A bite is the act of penetrating human sk in 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 tic k or chigger moves onto treated skin or by how long the tick or chigger remains on treated skin.

5(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 firs t event is confirme d by the second; the second event is the confirming event

.
(4) A human subject is a living individu al about whom an investigator conducting research obtains either data through interv ention 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 ch iggers 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 th em 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 failur e 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 protocol s 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 de velopment 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 requi rements of 40 CFR §26.1125, must then be reviewed by EPA and the HSRB befo re research is initiated.

It is critical to all later stages of repellen t testing that protocol s contain the elements required by EPA’s Good Labor atory Practices regulations at 40 CFR §160.120 and meet all other applicable scie ntific and ethical standards. The broad topics of scientific design, ethical justification, subject selection and in formed 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 re search 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 ques tion. The design should include a detailed description of the experimental desig n, addressing topics (i) through (x).

6(i) Objectives. The objective of most repellent efficacy testing to support registration is to estimate how l ong after treatment a repellent will continue to protect users from the ta rget pest; this period of effective repellency is measured as Complete Protection Time (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 Pr otection (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 propo sed for registration. Test materials should be stored at ambient temp erature 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. Cons iderations in endpoi nt 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/cm 2 of treated skin surface or volumetrically as ml/cm 2 of treated skin surface. Because each subject is likely to apply a di fferent amount of repellent in uncontrolled trials, the “typical dose ” should be calculated as the mean of multiple applicati ons by each of many subjects.

(B) Repellency. The endpoint of repellency testing should be selected to show a failure of repellent e fficacy 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 la nding, bite, or crossing confirmed within 30 minutes by another similar event. The Agency encourages the use of the first c onfirmed failure event to estimate complete protection time for indi vidual subjects, and the median CPT across all subjects in each treatment arm as the summary measure of CPT.

( 1 ) In repellency testing with mo squitoes or biting flies the study design should choose either landings or bites as the event showing failure of repe llent efficacy. The Agency encourages the choice of landings for field testing to reduce

7the risk of exposing subjects to vector-borne diseases. Even in laboratory testing us ing laboratory-reared, diseasefree insects, the choice of landi ngs reduces the risk of an adverse reaction to b ites. 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 repe llent 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 pr ovided by the repellent, and long enough that all or nearly all subjec ts 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 ca re in study design—for example, by planning a test of longer duration or , when appropriate, by pre-treating subjects well before their first expo sure to target pests. Methods for analyzing right-censored datasets ar e discussed in paragraph (c)(1)(xi) below.

(v) Sample size. The sample should be large e nough to be likely to yield a definitive answer to the research question being addressed, and its size should be justified statistically in ea ch protocol, taking into account the specific characteristics of the proposed research and the desired accuracy and precision of the resu lts. 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 co mpromise the validity or utility of test results. The protoc ol should fully describe how the proposed sample size was determined, and how data fo r subjects who withdraw from the test prematurely will be treated.

Other factors which may affect sample size are the numbe r of treatments, the experimental design, and the hete rogeneity of the target population (e.g., by age, sex) and of the environm ent (different habitats, different conditions, different species a nd population densities).

(vi) Allocation of subjects to treatments. Subjects should be allocated to treatments randomly and, to the extent possible, blindly.

8Multiple treatments per subject may be permissible if all treatments contain the same active ingredient at comparable concentrations, but caremust be taken in the study design and execution to prevent crosscontamination or confounding interference. It is particularly important for subjects receiving different treatment s 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 feedin g close to the ground on lower limbs, treatments should be applied only to subjects’ lower legs, an d only two treatments per subject will be possible. Treating th e 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 protecti on 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 untr eated 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 ade quate 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 de sirable 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 cm 2

. Positive controls are particularly valu able when the objectives of the study include comparisons between formul ations, and when testing spans different days. To minimize the like lihood 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 c ontrol, subjects’ limbs s hould be washed with an unscented detergent and carefully rinsed and dried. Subjects should avoid alcohol, tobacco, and scented product s (perfume, cologne, hair spray, lotion, soap, etc.) for at least twelve hours before and throughout the test. Subjects should avoid activities th at increase perspiration, and avoid abrading, rubbing, touching, or we tting the treated area.

(x) Treatment of subjects. Subjects in trials desi gned 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,

9aerosol spray, towelette, or lotion) a nd with the directi ons 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 a pplication and uniform coverage.

(xi) Statistical analysis plan. Protocols should include a full description, explanation, and justific ation for the statistical methods proposed to analyze both dose determination and re pellency test results, taking into account the specific study object ives and variables.

The statistical analysis plan should provide for testing results for normal distribution. When results are normall y distributed, it may be appropriate to report the mean CPT across all treate d subjects with its standard error. When the data do not fit a normal dist ribution—more typical of repellency datasets—it may be possible to transf orm 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, nonparametric analyses, such as Kaplan-Meier survival analysis 1 or maximum likelihood methods are suggested.

If the study objective is to determin e CPT across all subjects, any rightcensorship of repellency data will lead to underestimation of both the mean and the variance around it. Beca use 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 st udy 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, th e risks to subjects must be minimized for the research to be ethically justified. The expe cted benefits to society from the knowledge likely to be gain ed 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 ne w 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 Testi ng 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.

10(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 se nsitization studies are needed to estimate the margin of exposure (MOE ) for subjects participating in a repellent test. All additional componen ts 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 thei r 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 prot ocol. Potential risks in repellent testing typically include (but may not be limited to) reactions to the test substance or to arthropod b ites, acquisition of vector-borne illness, physical risks (e.g., stresses imposed by the requirements or conditions of the test), or po ssible 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 Institutional Review Board (IRB) 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 §26.1111(a)(1).) Different actions are effective in minimizing different risks; protocols should identify specific steps taken to minimize each identified risk to subjects.

Protocols should discuss sp ecific steps proposed to prevent research-induced harm—for exampl e, 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 inve stigator’s or sponsor’s committing to pay for uninsured costs of medical treatment of subjects for injuries or illnesses re sulting from their participation in the research, and for longer-term treatment, if needed, of any arthropod-borne disease cont racted during testing.

Study protocols should include pr ocedures for monitoring the safety of subjects, stopping ru les specifying conditions under

11which a subject would be withdr awn from the research or the 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 ar e minimized in repellent testing may involve some or all of the following:

• Monitoring of potential field te sting sites for the presence of disease vectors at least w eekly during the month before testing. Sites at which inf ected 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 determ ine 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 labor atory-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-inves tigators are present at all times so that the Principal Investigator can, if needed, attend to the safety of a s ubject without compromising the integrity of the research or endangering other subjects;

• Initiating post-study contact wi th 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 indi vidual subjects, yet is not risk-free, even after risks have been minimized. Thus the ethical justification for the research ultimately depends on the e xpected benefits to society of the information likely to be gained fr om 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 accr ue. In addition, protocols should estimate the likelihood of achieving each identified societal benefit.

12Societal benefits include only thos e 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 othe r 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 IRB must also determine th at minimized “risks to subjects are reasonable in relation to anticipated be nefits, if any, to subjects, and the importance of the knowledge that may r easonably be expected to result. In evaluating risks and benefits, the IRB should consider only those risks and benefits that may result from the research. . . . The IRB 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 §26.1111(a)(2).) The protocol should discuss explicitly the balance of risks to subjects and anticipated so cietal 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 cr itical to the generalizability of the results of research, and to be ethically acceptable, subject selection must be e quitable. (40 CFR §26.1111(a)(3)) Fully informed and fully voluntary consent of subjects is fundamental to ethical human resear ch. (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 targ et population of repellent users. Samples should include adults of vari ous ages, of both sexes, and of a variety of races, and protocols should describe the demographic characteristics of the pool from wh ich subjects will be recruited. Attractiveness of recru ited candidates to the ta rget species should be verified before they particip ate in repellency testing.

Testing with a sample known to be unrep resentative is discouraged; if it is proposed, it should be fully jus tified. Arguments based on the convenience of the investigators or on the difficulties associated with recruiting a more representative samp le do not justify testing with an unrepresentative sample.

(ii) Inclusion/Exclusion factors. All exclusion/incl usion criteria propose should be supported by a clear and exp licit rationale. Because testing a topical repellent necessarily involve s intentional exposure of subjects, children under 18 and pregnant or nurs ing women must be excluded. 40 CFR §26.1203.) Because older adults are more susceptible to arthropod-borne diseases, candidate s over 55 should be excluded as d

(See

subjects in field testing, or their inclusion should be specifically justified.

13Members of certain vul nerable popula tions, including people of limited mental capacity, those not in good hea lth or with compromised immune

y ulty

erally be recruite d from populations in the area where ting will be conducted. An of fering of distant travel may unduly

a

(iii) cruiting process, from the firs t contacts with potential candidates

ended

in

(iv) nsation of subjects. It is reasonable to compensate participants in pellent efficacy research for thei r time and trouble. The level of

ation

e y time

(v) h ith human subjects that participatio n of subjects be both fully informed

med consent is an extended process, involving much more than simply obtaining a subject’s signature on a form. systems, those sensitive to chemicals, and students or employees of the investigators or sponsor s should always be excluded. Other potential subjects who may be in a vulnerabl e position, including the educationall or economically disadvantaged, or th ose who communicate with diffic because of language differences or disabilities, should not be excluded arbitrarily if appropriate specific provision can be made to ensure their safety and welfare.

Subjects should gen
tes influence a candidate’s choice to en roll, and a subject who has accepted long-distance transportation may feel less than free to withdraw from study. If it is proposed, transportation of subjects to distant locations should be justified, and specific m echanisms should be proposed to prevent any coercion or undue influence.

Methods of recruiting. Protocols should describe in detail the proposed

re through all discussions of the research and the subjects’ enrollment. Any advertisements or flyers proposed for use in recruiting should be app to the protocol, and should reflect IR B 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 sp eak or read a language other than English, procedures for accommodati ng their needs in the recruiting process and in the conduct of the re search itself should be described detail.

Compe

re 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 particip in the research attractive only to the economically disadvantaged. Compensation should not be used or admi nistered so as to compromise th freedom guaranteed to subjects to withdr aw from participation at an for any reason, without sacrificing be nefits to which they are entitled.

Informed consent. It is a fundamental requi rement for ethical researc

w and fully voluntary.

(A) Process. Infor

14It begins with recruiting, and continues after signature of the consent form throughout the conduct of the research. A single discussion in the protocol of th e recruiting and consent proces as a continuous whole is recommended.

As a condition of approval of proposed res ses

earch, an Institutional Review Board (IRB) must determin e that informed consent will be

r ircumstances that provide the prospective subject . . . sufficient

t te, and

l to

e investigator ill confirm each candidate’s understanding of the research and of

quirements for formed consent, including at §26.1116(a) the basic elements

de

(B) he rule at §26.1117 provides two options for documenting informed consent. The first

ally 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 CF R §26.1111(a)(4) and (5).)

The rule at §26.1116 requires that consent be sought “only unde

c opportunity to consider whether or not to participate and that minimize the possibility of coercion or undue influence. The information that is given to th e subject shall be in language understandable to the subject.” EPA recommends that consen discussions with indivi dual candidates be conducted in priva that all consent documents be written at or below an 8 th grade reading level, as measured by the Flesch-Kincaid Grade Level score or another standard tool for assessing readabilit y. (A too calculate the Flesch-Kincaid Grade Level score is a standard feature available to users of MS-W ord.) 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 th

w what it will involve before he or she is enrolled.

The rule at §26.1116 further define s the general re in always required, at §26.1116(b) other elements conditionally required, and at §26.116(e) a requirement to identify the pestici and the nature of its pesticidal function.

Documentation of informed consent. T

option is a “written consent docum ent that embodies the elements of informed consent required by § 26.1116.” The second is “a short form written consent documen t stating that the elements of informed consent required by § 26.1116 have been presented or 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,

15it should be justified, and prior di scussion with EPA is strongly encouraged.

Ensure that an y elements appear ing in both the protocol and the consent form are in harmony. Elements common to both

es in the

om

orm should tell subjects how any bites, if any, they are likely to get, and what symptoms of ting

(4) Protection of nd confidentiality. It is important to protect the privacy and confidentiality of subjects in repellent efficacy research, and to

e in data collection forms or study reports. S ubject names unavoidably must appear

(ii) s. If photographs or videos are made to document the research, care should be take n to minimize making recognizable

he

(iii) ng of female candidates for pregnancy should be handled with care and discretion. The investigator is responsible for

ng documents may include, for example, descriptions of the purpose of the research, subject inclusio n/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 substa nce, although the presentations should usually not be the same, because the readers and purpos differ. Consent forms should addre ss candidates consistently second person—as “you”—and shou ld present all information from the subject’s point of view. By contrast protocols should be written in the third person, and s hould present all information fr the investigators’ point of view.

In repellent testing, the consen t f

m arthropod-borne disease they should be alert for after participa in a field study.

subj ect privacy a

inform candidates in the recr uiting 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 nam

on consent forms and on administrativ e 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.

Photographic image

images of subjects. If faces or id entifying marks cannot be excluded from a photograph or video image, the imag e should be altered to protect t identity of the subject(s).

Pregnancy testing. Testi

ensuring that pregnant or nursing wo men are excluded as subjects (See 40 CFR §26.1203). It is not necessary to meet this responsibility by testi all female candidates for pregnancy. Fo r example, it is neither respectful nor informative to require women wh o are post-menopausal or surgically

16incapable of pregnancy to take a pr egnancy test. Investigators should consider how best to meet their re sponsibilities while fully respecting female subjects.

When pregnancy testing is conducted, the circumstances of testing should protect the privacy of the candidate, bo th during the testing itself and in

ecruit

(d) eview of protocol s for repellent studies. EPA’s Rule for the Protection of Human Subjects of Research requires ex tensive review of protocols be fore research is initiated.

criteria for IRBs are defined in the rule at 40 CFR §26.1107-§26.1111. Records handling the results. Unexpected news of a pregnancy can cause significant psychological distress, wh ich can be increased by any breach of discretion in handling the informa tion. It is a good practice to r 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 record s of a positive pregnancy test should be retained.

R

The rule requires that the complete protoc ol, 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, ge neral procedures, and decision

to be maintained by IRBs are define d 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 e in the otocol

(ii) a for IRB approval of proposed research are defined in the rule at 40 CFR §26.1111. In summary, this h

are minimized;

• Risks to subjects are reasonable in relation to the importance of the expected to result from the research; complete protocol, a consent form, and all materials intended for us recruiting process or to be provided to candidates or subjects. A pr responsive to section (c) of this guidelin e is likely to satisfactorily address most requirements of a reviewing IRB.

Criteria for IRB approval. The criteri

passage of the rule requires that the IRB determine that proposed researc meets all these criteria:

• Risks to subjects

k nowledge that may be reasonably

17• Selection of subjects is equitable;

• Informed consent will be sought from each prospective subject;

• Informed c onsent will be appropriately documented; ion is made for monitoring

ect the

included in the study design to pr otect their rights and welfare.

(iii) ve, arch proposals. (40 CFR §26.1109.) It is common for IRBs to grant

the r of
tors or

(2) Review protoco iew by EPA and by the HSRB. (See 40 CFR § 26.1125.)

efined in the rule at 40 CFR §26.1125 and summarized in Appendix A to this guideline. Records of

(a)–(d) call for a discussion of the risks and benefits of the proposed resear ch, for the consent forms as
ut

ation to

this

n the IRB and the investigators or sponsors (§26.1125(e)) and official notificatio n of IRB review and approval • When appropriate, adequate provis

c ollected data to ensure th e safety of subjects; and

• When appropriate, adequate provision is made to prot
p rivacy of subjects and to mainta in the confidentiality of data.

I n addition, the IRB ensures that when any subjects are likely to be vulnerable to coercion or undue influence, additional safeguards are

Documentation of IRB approval. IRBs have the authority to appro require modifications in (to secure approval), or disapprove rese

conditional approval subject to the investigator’s making specified modifications in the proposal; once required modifications are made, IRB will issue formal written approval of the research. This lette approval, along with all other correspondence betwee n the investiga sponsors and the reviewing IRB, must be included in the subsequent submission to EPA. (See 40 CFR § 26.1125(e) and (f).)

s by EPA and the HSRB. After receiving approval from an IRB, the l and supporting material must be submitted to EPA for rev

(i) Required elements in submission to EPA. The elements to be included in a protocol submission to EPA ar e d

two kinds are required.

(A) Records describing the research proposal itself. 40 CFR sections 26.1125

submitted to and as approved by the IRB, for information abo how subjects will be recruited, and for a description of the circumstances and methods proposed for presenting inform subjects and seeking their consent. A protocol responsive to the guidance in section (c) of this gui deline will already include range of information.

In addition, 40 CFR §26.1125(e) and (f) call for submission of all correspondence betwee

(§26.1125(f)). For purposes of §26.1125(e), “correspondence” does not include attachments transmitted by correspondence, such

18as the protocol or consent form, wh ich are otherwise required to be submitted by other provisions of the rule.

Records of the IRB review. 40 CFR §26.1125 also requires submission of copies of records re levant to (B) the proposed research which the IRB is required to maintain by 40 CFR §26.1108(a). ed

w gainst, and abstaining; the basis fo r requiring changes, and

d degrees, fficient to describe each member’s chief anticipated contributions

cribed nd §26.1108(b).

it them directly to EPA.

(ii) EPA Review. quired by the rule at 4 satisfaction of the requirements of 40 CFR §26.1125, and for substantiation of any

al

call of the proposal before it is reviewed by the HSRB. If EPA determines that the proposal is ready fo r HSRB review, it will be scheduled for

he HSRB There is some overlap between these records and those discuss above, but there are some im portant differences. 40 CFR §26.1108(a) calls for, among other records:

• Minutes of IRB meetings, in sufficient detail to sho attendance; actions taken; votes for, a

s ummarizing the discussion of controverted issues;

• A list of IRB members identified by name, earne representative capacit y, indications of experience su

t o IRB deliberations, and any employment or other relationship between each me mber and the research institution; and

• Written procedures for the IRB in the same detail des in §26.1108(a) a

T hese records can only be obtaine d from the IRB. By prior arrangement, however, an IRB may subm

Timely EPA review of submitted proposals for research is re 0 CFR §26.1601. EPA will first review submitted proposals for

claims of business confidentiality associat ed with the submission. If a submission is found to be incomplete, or to incl ude unsubstantiated claims of business confidentiality, the submitter will be prom ptly 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 ethic 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 for revision

HSRB review at the earliest opportunity. The submitter may respond to EPA’s review in writing to the HSRB docket, a nd may also comment orally at t meeting.

19(iii) of the protocol by Agency sta ff, EPA shall submit the protocol and all supporting materials, together with the staff evaluation, to the Human Studies

e

ublic

rch HSRB Review. As provided in 40 CFR §26.1601(d), “following initial evaluation

Review Board.” The HSRB is an advisory committee, chartered under th Federal Advisory Committee Act, charge d with advising EPA on scientific and ethical aspects of proposed and complete d research with hum an subjects. The HSRB meets several times a year in pub lic session, with opportunities for p 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 resea 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

.

es to IRB-approved research before execution. After EPA and the HS (e) Chang RB have reviewed and commented on proposed research, revisions to the prot ocol, consent form or other materials may be needed before th e research is executed. When IRB-approved

RB.

e

d

(f)

(1) Execution of protocol. When EPA and HSRB reviews are completed and the isions resulting from those revi ews, the research can be initiated. Subjects can be recruited and da ta can be collected as specified in the in

(2)

Practices regulations at 40 CFR part 160. These rules require that each testing col and .35) materials are revised prior to in itiation of the research to ad dress concerns raised in EPA and/or HSRB reviews or for any other reas on, those revised materials must be resubmitted 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 IRBapproved consent form, and to all changes to any other materials approved by an I (40 CFR §26.1108(a)(4)). Although an IRB has the discretion to conclude that som 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 propose research which are implemented w ithout prior approva l by the IRB.

Execution of repellent studies

IRB has approved any rev

protocol. Care should be taken to ensure and document that these events occur the proper sequence. Initiation of subject recruitment before obtaining IRB approval of the final protocol will be treated as a breach of the regulations.

Quality Assurance (QA) oversight. Repellent efficacy testing, whether conducted in the field or in the labora tory, is subject to the Good Laboratory

facility include an independent QA unit to monitor execution of each proto document its conduct in accordance with the GLP regulations. (40 CFR §160

20(3)

All such deviations from the protocol should be promptly and fully reported to the cussed

4))

(4) ed to the IRB for review, and must be appr oved by the IRB before it is implemented,

(5) ly reported to both the overseeing IRB and to EPA, including adverse events not esta blished to be related to product exposure

(g) Report

(1) Study report. In addition to the standard elements required by the GLP the append ices described below, the primary report of the study should include these elements, as applicable.

cility, and dates of the study.

ii) Recruiting and enrollment st atistics, including the following:

• Demographic characteristics of th e pool from which subjects were

Numbers of subjects enrolled, withdrawn voluntarily, withdrawn

(iv) age or field site. For laboratory studies, the source of test ar thropods, methods used to establish Deviations from protocol. In executing even the best designed and most comprehensive protocols, unanticipated devi ations from the protocol may occur.

IRB, and both the deviations and their im pact on the research should be dis in the study report submitted to EPA. (40 CFR §160.185) EPA makes a distinction between planned changes to a pr otocol and deviations from a protocol. Planned changes must be treated as amen dments to the protocol, and must be approved by the IRB before they ar e implemented. (40 CFR §26.1108(a)(

Changes in IRB-approved research after initiation. Any amendment to the protocol, consent form, or other materials approved by the IRB must be submitt

unless immediate implementation is requi red to avert an imminent hazard to subjects. (40 CFR §26.1108(a)(4))

Reports of adverse events. Any adverse events a ffecting the subjects of a repellent efficacy test must be prompt

or study participation. (40 CFR §26.1108(b)(1))

ing of completed repellent studies

regulations at 40 CFR §160.185 and

(i) Study identification: Title, id entifying study number(s), sponsor, study director, investigators, name and loca tion of the testing fa

(i i) Approved or intended label directions for use.

(i

recruited and of the s ubject sample selected;

• Numbers of candidates contacted, interviewed, and screened; and

• i nvoluntarily, and completing the research.

Test arthropods: Genus, species, su bspecies or strain (if information is available) of target pests; and pest pr essure in each c

21and maintain their disease-free stat us, rearing techniques and conditions developmental stage, age and sex of target arthropods and methods used to sex them; preparation of arthropods befo re testing, and density per cage or container.

Test conditions: For field studies, the location and type of habitat, species distribution ,

(v) , climate and environmenta l conditions. For laboratory studies, a complete description of cages or containers, temper ature, relative

• Preparation of subjects: Traini ng, demonstrating attractiveness, n, calculation of skin area and individual dose, washing, rinsing, and drying of limbs;

• Method of application of repellen t to each subject and individual

• Time of start and end of exposure of each subject.

(vii) Complete accounting for all events during the testing period.

(viii) esults for each subject.

x) Reports of all statistical analyses

) Conclusions

(xi) Discussion

(2) Appendices.

(i) Complete protocol as approved by the IRB.

(ii) Consent form and any additiona l recruiting materials as approved by the hum idity, ambient light, and air flow.

(vi) Test procedures, including but not limited to:

clothing and protective equipment wor

• Allocation of subjects to treatm ents; number of treatments per subject; blinding key, if used;

• Method of dose determination;

d ose applied;

• Time of application of repellent to each subject;

R

(i

(x

IRB.

22(iii ) All amendments to the protocol, consent form, or other materials as
appro ved by the IRB.

pact esearch.

i) Documentation of ethical cond uct of the research, as required by 40 CFR ce with IRB not previously submitted to EPA. The requirements of this se ction of the rule are summarized in

(h) Retention of R -keeping requirements apply to some or all records of research covered by this guideline.

an research conducted under EPA’s Human Research Rule.

(2) who h with hum ans subject to FIFRA §12(a)(2)(P).

nce or tolerance exemption or in support of registra tion or an application for registration.

(4)

(i) Specifi repellency testing should be treated with th e test material at a standard dose rate,

xpressed by weight of repellent per unit area, mg/cm 2 of treated skin, or volumetrically depend ng of

d

in (iv) Reports of all deviations from the protocol and assessment of their im on the integrity of the r

(v) Raw data and data collection sheets.

(v §26.1303, including all corresponden

Appendix C to this guideline.

ecords. The following record

(1) The record-keeping requirements in 40 CFR §26.1115 apply to Institutional Review Boards (IRBs) that review hum

The record-keeping requirements of 40 CFR §169.2(j) apply to investigators conduct pesticide researc

(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 tolera

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.

c guidance for dose-determination studies. All subjects participating in

e as ml/cm 2 of treated skin. Recommended methods for choosing the standard dose in part on the active ingredient(s ) and formulation(s) of the te st material. For testi lotion formulations containing deet, a standard dose rate of 1 g per 600cm 2 (equivalent to 1.67 mg per cm 2 ) 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 empirical ly through a dose-determination study. For other ingredients and formulati ons no comparable standard dose has been identified, an it is recommended that a “typical consumerdose” be estimated through empirical dosedetermination testing as described below. The rationale for the dose rate actually used repellent testing should be docum ented in all study reports.

23(1) Test material. The formulated product should be used as it is or will be marketed, in the same type of container and bearing t he same directions for use.

formed

participation. The same or very simila r eligibility criteria would apply as for

ed with ld

(3) an be ntimeters at the wrist and elbow, or of the leg at the ankle and knee, and in either case at one or

or

(4) ally would apply a repellent to achieve complete c overage. Once the quantity applied has

by
towe lette before and after use. Because less than all the spray released from a pump-spray or aeros ol container will reach the target limb,

n per cm

(5) and the grand mean across all subj ects of mean doses applied to forearms and to lower legs s hould be calculated. The specific gravity (2) Subjects. Subjects used for dose determinati on should be recruited and in as described above, and mu st provide written informed consent for their

repellency testing, although subjects pa rticipating only in dose-determination testing would not be exposed to any insects or to any of the risks associat exposure to insects. The number of s ubjects used for dose determination shou be justified statistically. The sample should include adults of both sexes and varied race, and should be as representa tive as possible of the repellent user population, consistent with ethica l and feasibility constraints.

Measuring subject’s skin area. The surface area of subject’s limb in cm 2 c estimated by measuring the circumference of the forearm in ce

two equally spaced intermediate poi nts; then multiplying the average circumference by the length of the limb in centimeters from the wrist to elbow from the ankle to the knee.

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 norm

been measured and recorded, the applied repellent should be washed completely off the limb. This process should be re peated at least 3 times for each limb treated by each subject.

The applied quantity of a lotion or towe lette formulation can be determined weighing the container or

a different method is needed to esti mate the applied quantity of a spray formulation. The target limb should be wrapped in gauze “b racelets” of known area so that the bracelets c over only part of the skin. So long as the area of the bracelets is known, the quantity of a spray formulation applied to the ski 2 can be estimated by weighing the bracelet s before and after application. Skin should be left exposed between bracelets to help subjects determine when they achieve complete coverage. Dose determ ination with spray formulations should be conducted out-of-doors.

Calculating standard dose for use in repellency trials. The mean dose applied by each subject to each limb

of the test material should be used to convert the dose expre ssed by weight as mg
per cm 2 to a volumetric dose expressed as ml per cm 2 ; this calculation should be included in the study report. The volumetri c dose should be used as the standard dose for repellency testing, scaling it to the treated surface area of each subject’s

24limb or limbs. The protocol should incl ude the specific gravity of all test materials and should specify this calc ulation, and should specify doses both by weight and by volume.

c guidance for labo rato (j) Specifi ry studies of mosquito or biting fly repellency.

(1) Species. Mosquito tests should be conducted with Aedes aegypti , an Anopheles

quinquefasciatus. Stable fly tests should be conducted with Stomoxys calcitrans

.

s 5- uld be reported. Stable flies should be male or female adults 3-10 days old. The age range of test insects should be reported.

0% ± 10%, and photoperiod 16:8 hours (light:dark). Alterna tive rearing methods for

before use in a test, and star ved for 12 to 24 hours immediately before the test. Test stable flies may be fed dry sugar cubes w ith a separate water source; ved

to 232,000 cm ), square or rectan gular in plan, with one or more sleeved openings for the s ubject’s arms. A mirro r on the bottom of

s.

l.

each 4,640 cm .) 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. species such as An. albimanus , or a Culex species, preferably Cx.

The source of test insects should be re ported, and if possible, they should be identified by subspecies or strain.

(2) Stage, age, and sex. Mosquito testing should be conducted with adult female 10 days old; methods of sexing sho

(3 ) Rearing techniques. Mosquito and biting fly la rvae should be reared under optimal conditions for the species—typically at 27° ± 3

 ° C, relative humidity 8

mos quitoes are discussed by Gerberg et al . (1998). Use of alternative rearing techniques for biting flies and/or mosqu itoes should be fully documented and justified.

(4) Preparation of insects. Test mosquitoes should be fed 10% sucrose and no blood meal

the y should also receive no blood meal before use in a test, and should be star for 24 hours immediately before the test. Te st 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 3

the cage helps observers see insects landi ng on the underside of subjects’ arm 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 detai

(7) Insect density. There should be at least 200 mo squitoes in each 2’x2’x2’ cage (equivalent to one for each 1,160 cm 3 ) or at least 50 stable fl ies per cage (one for
3

25(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.

nd

tion of ethanol or isopropyl al cohol in water, and dried with a clean towel. The surface area of each test subj ect’s treated forearm (in

ts or treat ed subjects; multiple untreated controls should be used as a standard of co mparison only in studies of relative d

m 2

2) Establishing subject attractiveness to test insects. Before the test, subjects

less or 2 stable fly landings in two minutes or less are sufficient to establish attractiveness.

ed

removed from the subject’s arm, by shaki ng it or by other means, before they

e e, and Culex spp. When testing in subdued light , care should be taken not to compromise the investigators’ ability to observe insect activity. Investigators a subjects should avoid exhaling into the test cage; introduction of CO 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 solu

cm 2 ) 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 subjec should not be compared to those f

prot ection/percent repellency. To mi nimize potential interference, untreate 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 c with 20% deet in ethanol is recomm ended to calibrate the test system.

(1 should expose their untreated forearms to the target insects in a test cage to establish their attractiveness. Five mos quito landings in one minute or

(13 ) Establishing and confirming landing pressure. Before exposing any treat subjects during each exposure period, an untreated subject’s forearm should be inserted into each cage to confirm landi ng pressure. Landing insects should be

have time to bite. The untreated subject should remove his or her forearm from the cage as soon as it has received the requi site number of landi ngs. If at any tim fewer than 5 mosquitoes land on the untreat ed control forearm within one minut all mosquitoes should be removed from a ll 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 w ithin two minutes, all stable flies should be removed

26from that cage and 50 fresh flies should be added. The aggressiveness of the fre insects should be confirmed before repellency testing continues.

Exposure period. Approximately 30 minutes after treatment with the repellent, test subjects should insert th eir treated forearms into th e cage for sh

(14) the first time. Exposures of pre-defined duration should be repeated at regular intervals—e.g., 5-

r

(k) Specifi d studies of mosquito or biting fly repellency.

(1) Pre-test subject preparation. Subjects in field studies of repellency should be target species and to use aspirators to collect landing insects before they have time to

(2) ted in at least two di stinct habitats (e.g., forest, grassland, salt marsh, wetland, beach, barns, or urban environments) where the predominant mosquito is

o minimize risks to subjects, field testing should not be conducted where West Nile virus (WNV) or other mosquitole

(3) e ance of the udy for U.S. regulatory purposes should be justified in the study report.

ld The

organisms. The results of these analys es should be reported to subjects and included in the study report. minute exposures at 30-minute intervals—until efficacy failure for each subject o the end of the study, whichever occurs first. Subjects should avoid rubbing their arms when inserting them into or re moving them from the cage and between exposure periods.

c Guidance for fiel

trained in the laboratory to identify la ndings, probes, and bites of the

bite.

Choice of field testing sites. Field tests for mosquito repellency should be conduc

species differ. So long as the environmen ts and species distribution differ, it not important for test sites to be wide ly separated geographically. Repellency against black flies, stable f lies, or other biting flies t ypically 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. T

vectored diseases have been detected w ithin the previous two weeks. Because biting flies are not known to vector human diseases in the USA, no comparab site monitoring is required for studi es of repellency to biting flies.

Species identification. Mosquito tests should be conducted where more than on species is present. If tests are conduct ed outside the U.S., the relev
st Landing insects should be aspirated or trapped before and during the test, and labeled with the time of collection. Af ter the field study, collected insects shou be identified by genus and sp ecies, and if possible, by s ubspecies or strain. number in each taxon collected in ea ch time period should be reported.

After identification, mosquitoes should be subjected to se rological or other analysis to determine the presence or absence of WNV or other disease

27(4) y d be ith a repellent on any limb.

sure. Results for the untreated controls should not be compared to results for treate d subjects. When
ects are

(5) itive controls. A positive control group treated at a rate of 1 ml per 600 cm 2

(6) Minimum landing pressure to initiate or continue test. Landing pressure

or continued unless landing pressure of the ta rget species is at least one mosquito

or

ler treated area on the appropriate limb of at least 250 cm is also acceptable. The

s’ ugh

(7)

o subjects and al low them to rest between exposures.

the

Reliable results may be obtained for ex tended periods of protection by treating subjects up to several hours before the first field exposure, with subsequent Untreated controls. Untreated control subjects are necessary in all stud designs. To minimize the potential for inte rference, untreated c ontrols shoul subjects who are not treated w

When the objective is to estimate CPT, two untreated cont rol subjects are sufficient. They should expose an untreate d arm or leg briefly at regular intervals during the test to confirm continued accep table landing pres

the objective is to evaluate relative prot ection, more untreated control subj needed to provide a standard of statistical comparison.

Untreated limbs used as controls should be washed, rinsed, a nd dried exactly like treated limbs.

Pos with 20% deet in ethanol is recomm ended to calibrate the test system.

sho uld be measured before treatment and intermittently throughout the course of the test by untreated control subjects . Testing should not be conducted

lan ding within one minute, or at least one stable fly, black fly, ceratopogonid tabanid landing within five minutes. In sects landing on untreated subjects should be collected for later identification, a nd 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 lo wer leg (ankle to knee), depending on the feeding behavior of the predominant species at the selected test site. A smal
2 treated area should be washed with uns cented 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, subject heads, trunks, and limbs should all be c overed with light-colored material thro which insects cannot bite.

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 t

When testing a repellent with a long period of effectivene ss, it is critical to 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.

28exposures timed to coincide with periods of target insect activity. This approac 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 exposur and then exposing all subjects at the same time when target insects are active. h

e)

(8) Minimizing variation. Many factors contributing to variability in repellent field

allocation to treatment, subject attractivene ss to target insects, etc. Other factors

tivity

(9) at

. ty, and wind speed) should be monitored periodically throughout the study and re ported. Testing should be not be

(10) each

ubjects affect test results.

collection.

(l) Specifi reliable cy against ticks and chiggers in the laboratory. If field te sts are conducted, proposed label

irections for re-application frequency should reflect field study results.

f
at an angle of 30° or more to the surface. With a suitable instrument (such as an studies cannot be controlled, but mi nimizing variation when possible and designing studies with insect feeding patterns in mind can make a study design more efficient. Standardization is possi ble for such factors as time of testing,

which may be managed to minimize vari ation include target ing a predominant species in its habitat, synchronizing expos ure periods with peak feeding ac of target insects, and treating limbs consis tent with the preference of the target species to feed on legs, arms, or both.

Environmental conditions. The time of day at which subjects are treated and which exposure to target insects begins a nd ends should be recorded and reported Weather conditions during testing (inc luding temperature, relative humidity, cloud cover, precipita tion, light intensi

conducted or continued if wind speed exceeds 16 kph/10 mph.

Test subject pl acement and behavior. Subjects may work in pairs to assist other to identify landings a nd collect insects. Becaus e clustering of subjects may confound results, at least 3 m/10 ft should be maintained between pairs. S should avoid strenuous exercise and sweating, since these coul d

(11) Data collection and reporting. Under the supervision of the investigator, an associate or another subject should reco rd 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 identif ication and labeled with the time of

c guidance for laboratory st udies of tick and chigger repellency. Because field tests have not been developed, EPA recommends testing for repellen

d

(1) Summary of recommended method. Each subject should place the fingers o one hand on a flat surface, with the elbow above the wrist and the forearm held

29artist’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 m arked 3 cm below the boundary of the treated area of the forearm. The tick or chigger should

ot

t 3 cm

e treated area have worked effectively in tests with nymphal Dermicentor variabilis and Ixodes scapularis, and are provided here as examples.

(2) tick

m ),

tick, Ornithodoros turicata ). Chigger tests should be conducted using laboratory E. f

,

(3) ly

or be reported.

) Preparation of test arthropods . Ticks and chiggers should be disease-free. The d be oriented gently (e.g., with paint brus h, 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 ch igger that crosses at least 3 cm into the treated area (toward the elbow) is reporte d as ‘not repelled’. One that does n cross into the treated area, or that crawls into the treated area but immediately turns back or falls off, is reported as ‘repelled.’ Fresh tick s 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 operati onalized in each protocol in a way appropriate to the species and life stage of ticks to be tested. A release poin distant from the treated area and scoring of a crossing when a tick moves at least 3 cm into th

Species. Tick tests should be conducted us ing laboratory colonies of the 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, Ixodes pacificus ), lone star tick (Amblyomma americanu American dog tick (Dermacentor variabilis ), and relapsing fever tick (softbacked

colonies of chiggers in the Trombiculidae family; Eutrombicula splendens; or cinnabarrs . Test arthropods should be iden tified by genus and species, and i possible by subspecies or strai n. It is permissible to te st two species concurrently alternating so that each s ubject qualifies and tests a tick or chigger from each species within each exposure period.

Stage, age, and sex. When testing with blacklegged (deer tick), lone star, or softbacked ticks, either adult or nymphal life st ages are appropriate for testing. On the adult American dog tick is recommende d, since nymphs of this species do not feed on humans. Tests with chiggers s hould use immature chiggers. The age age range of all test ar thropods should

(4) Rearing techniques . Ticks and chiggers shoul d be reared at 25° ± 3 ° C, at high relative humidity (> 90%), and photop eriod of approximately 16:8 hours (light:dark). Any alterna tive rearing techniques shoul d be fully described and justified.

(5 source of test animals and the methods used to ensure they are disease-free shoul be reported.

30(6) n of test arthropods . Each tick or chigger should be used only once and should be destroyed immediately after use.

(7) ency one at a time, accord ing to the study design. Some studies using a single species of tic k test 5 ticks in each 30- minute exposure period; other

he

(9) e treatment, both forearms of each subject are washed with unscented detergent and caref ully rinsed and dried. One forearm

3 cm distant from the boundary line, outside the treated area toward the fingers. Similar lines ar e drawn in the same positions on the wrist

(10)

or 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 and

iate

(11)

(12) ne fresh immediately af ter demonstrating active questing on the subject’s untreated arm, is placed at th e release line on the treated arm. A

r at least one minute.

(13)

, Dispositio

Number of ticks or chiggers. Ticks or chiggers are te sted for questing behavior and for repell

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 maintain ed during the test at 20° to 25 ° C, with relative humidity at ≥ 35% and indirect ambient light 50 to 80%. T lights should be kept on.

Subject preparation . Befor

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’—

of the subject’s other, untreated arm—one line at the wrist, and another 3 cm away toward the fingers.

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 repe llency testing. W ith an appropriate instrument, such as an artist’s paint brus h, forceps, or a cotton swab, each tick chigger should be picked

or chigger that moves steadily from the release line across the boundary line upward along the subject’s unt reated forearm is activel y questing, and appropr for use in repellency testing. A tick or chigger failing this test should be immediately destroyed.

Positive controls. A positive control group treated at a rate of 1 ml per 600 cm 2 with 20% deet in ethanol is recomm ended to calibrate the test system.

Test procedure. With the treated forearm held upr ight above the table, o tick or chigger at a time,

“crossing” is recorded if the test organi sm crosses the boundary line at least 3 cm into the treated area within 3 minutes, and remains in the treated area fo

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

31untreated arm and tested for active questing. Qualified ticks or chiggers are then placed at the release point on the treated ar m and exposed to the treated area. The cycle continue s until efficacy failure or the end of the study, whichever occurs first.

(14) ticks or

r ubject.

(m) Refere line.

(1) d Testing Topical Applica tions of Compounds as Repellents for Medically Important and Pest Arthropods (Including Insects, Ticks and Mites): I Mosquitoes.

(2) American Society for Testing and Mate rials. (2003) E 1488-02 Standard Guide

osquito repellents. In Insect Repellents: Principles, Methods and Uses . Debboun, Frances, and Strickman, eds. CRC Press. (495 p.)

cy of IR 3535, KBR 3023, para-menthane-3,8,-diol, and deet to Black Salt Marsh

(5) Barnard, D.R. (1998) Mediation of D eet repellence in mosquitoes (Diptera:

uito size effects on repellent tests. Med. Vet. Entomol. 12:39-45.

(7) factors that affect their performance. In Insect Repellents: Principles, Methods

(8) P. (1994) Response of a chigger, Eutrombicula hirsti (Acari: Trombiculidae) to repellent and toxican t compounds in the laboratory. J. Med. Entomol. 31(4): 628-630. Data collection and reporting. Recording of crossings and repelled test chiggers arthropods during the expos ure period should be supervised by investigators. The investig ator or, an associate, or another subject should record the number and timing of all events occu rring during in each exposure period fo each s

nces. The following publications were consu lted in developing this guide

American Society for Testing and Mate rials. (2006) E 939-94 Standard Test Method of Fiel

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 m

(4 ) Bernard, D. R., Bernier, U. R., Posey, K. H. and Xue, R-D (2002). Repellen

Mos quitoes (Diptera:Culicidea) in Everglades National Park. J. Med. Entomology39(6): 895-899

Cul icidae) by species, age, and par ity. J. Med. Entomol. 35(3): 340-343.

(6) Barnard, D.R.; Posey, K.H.; Smith, D.; and Shreck, C.E. (1998) Mosq density, biting rate and cage

Carroll, S. P. (2007) Chapter 12: Eval uation of topical insect repellents and

and Uses . Debboun, Frances, and Strickman, eds. CRC Press. (495 p.)

Frances, S.

3233 f

s.

ito

1) Govere, J.M.; and Durrheim, D. N. (2007) Chapter 8: Techniques for evaluating
es,

2) Huntsberger, D. and Billingsley, P. (1981) Elements of Statistical Inference. 5 th

3) Klun, J.A. and Debboun, M. (2000) A new module fo r quantitative evaluation of

(14) R. (2007) Fiel d evaluation against mosquitoes of regular and polymer-based deet formulations in Manitoba, Canada,

5) Smith, C.N. (1955) Insect repellents. Quarterly Repor t, Entomological Research.

6) Verwey, R.E. (1996) Laboratory method for testing insect repellents on human red

7) WHO (2009) Guidelines for Efficacy Testing of Mosquito repellents for Human

8) WHO (1996) Report of the WHO Informal Consultation on the Evaluation and (9) Frances, S.P.; Eikarat, N.; Sripongsai , B.; and Eamsila, C. (1993) Response o Anopheles dirus and Aedes albopictus to repellents in the laboratory. J. Am. Mo Con. Assoc. 9(4): 474-476.

(10) Gerberg, E. J. ; Barnard, D.R.; a nd Ward, R.A. (1998) Manual for Mosqu Rearing and Experimental Techniques. AMCA Bull. 5.

(1 repellents. In Insect Repellents: Principles, Methods and Uses . Debboun, Franc and Strickman, eds. CRC Press. (495 p.)

(1 edition. Allyn and Bacon. Inc., Boston.

(1 repellent efficacy using human subjects. J. Med. Entomol. 37(1): 177-181.

Schofield, S.; Tepper, M.; and Gada wski,

with comment on methodological issues. J. Med. Entomol. 44: 457-62.

(1 Entomology Research Branch, U.S. Department of Agriculture. 8 p.

(1 test subjects against chig gers in the laboratory. Unpublished document prepa by S.C. Johnson & Sons, Inc., Racine, WI. 3 p.

(1 Skin: CDS/NTD/WHOPES/2009.4.

(1 Testing of Insecticides: CTD/WHOPES/1C/96.1.Appendix A: Checklist of El ements Required by 40 CFR §26.1125

34 40 CFR 26.1125 Prior submission of proposed human research for EPA review
Any person or institution who intends to conduct or sponsor hu man 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 addition al information, to the extent not already included:

Requirement Y/N Comments/Page Refs (1) Copies of arch proposals reviewed by the IRB, ic evaluations, if any, that accompanied the proposals reviewed by the IRB,

• progress reports submitted by investigators, and reports of injuries • all rese

• scientif

• approved sample consent documents,

t o subjects. (2) Minutes of IRB meetings . . . in sufficient detail to show

• attendance at the meet

• actions taken by the IRB; ings;

• umber of members voting

• the basis for requiring changes in or disapproving research; the vote on these actions including the n for, against, and abstaining;

• a written summary of the discus sion of controverted issues and their resolution. (3) Records of contin uing review activities. (4) Copies of all correspondence between the IRB and the investigators. (5)

• A list of IRB members identified by name; earned degre es; representative capacity; indications of experience such as board fficient to describe each member’s berations;

• ship between each member and fu ll-time employee, a member of governing panel or board, stockholder, paid or unpaid consultant. c ertifications, licenses, etc., su chief anticipated contributions to IRB deli any employment or other relation the institution, for example,

( 6) Written procedures for the IRB in the same detail as described in §26.1108(a) and §26.1108(b).All information relevant to the proposed research specified by § 26.1115(a)

ngs provided to subjects, as required (7) Statements of significant new findi by §26.1116(b)(5). ( 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 mean s of obtaining information comparable to what would be collected through the proposed research; and§112 )on

posed research.5(aa discussiof:

(5) The balance of risks and benefits of the pro §1125(b): All information for subjects and w ritten informed consent agreements as originally provided to t he IRB, and as approved by the IRB. §1125(c): Information about how subjects will be recruited, including any advertisements proposed to be used. §1125(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.The following Information, to theextent not already incl

§1125(f): Official notification to the sponsor or investigator . . . that research involving human subjects has been reviewed and approved by an IRB.uded:Appendix B: Framework for Science and Ethics Reviews of Proposed Human Research

35 tocol Review

(c) Principal Investigator and any sub-investigators

(e) Sponsor

n does it address? Why is this question important? Would the

f s , h ? question be answered without ne wly exposing human subject s? If so, how?

2. Stu

(a) W n explic hypothesis, what is it? (b) t this hypo he s?

2.1 Statistical Design

ar e proposed? Are proposed controls appropriate for the study design and st atistical analysis plan?

(d) What is the plan fo r allocating individuals to treatment or control groups?

(f) What is the plan for statis tical analysis of the data? the research question? (h) Does the proposed design have adequate statistical power to definitively answer the

ce of test material and formulation? the choice of dose/exposure levels and the staging of dose administration? (d) What duration of exposure is proposed? Q uestions to be Addressed in EPA Pro

Protocol Identification

(a) Title (b) Date

( d) Participating Laboratories

(f) Reviewing IRB

1. Societal Value of Proposed Research

(a) What is the stated purpo se of the proposed research? (b) What research questio research fill an important gap in understanding? (c) How would the study be used by EPA? (d) Could the research question be answer ed with existing data ? I (e) Could the o ow

dy Design

hat is the scientific objective of the study? If there is a Can the study as proposed achieve th at objective or tes it
t si

(a) What is the rationale fo r the choice of sample size? (b) What negative and positive controls

(c) How is the study blinded?

(e) Can the data be statistically analyzed?

(g) Are proposed statistical methods appropr iate to answer

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 choi (c) What is the rationale forAppendix B: Framework for Science and Ethics Reviews of Proposed Human Research

36 2.3 Endpoints and Measures

be m easured? Are they appropriate to the question(s) being

curate and reliable?

How will uncertainty be addressed? Will reported point values be accompanied by

measures of uncertainty?

3. Subject Selection

(a) What is the population of c oncern? How was it identified? (b) From what populations will subjects be recruited? (c) Are expected participants representative of the population of c oncern? 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/ex clusion criteria? Are they complete and appropriate? (b) What, if any, is the relationship betw een the investigator and the subjects? (c) If any potential subjects are from a vulne rable population, what is the justification for including them? (d) What process is proposed for recr uiting and informing potential subjects? (e) If any subjects are potentially subject to coercion or undue influence, what specific safeguards are proposed to prot ect 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 attrac tive 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 asso ciated with the research? How was this probability estimated? (a) What endpoints will asked? (b) What steps are proposed to ensure meas urements are ac (c) What QA methods are proposed? (d)

3.1 Representativeness of SampleAppendix B: Framework for Science and Ethics Reviews of Proposed Human Research

37 4.2 Risk minimization

(a) What specific steps are proposed to m nimize risks to subjects? sed dose/exposure levels co mpare to established NOELs/NOAELs for the test materials? hat stopping rules are proposed in the protocol? w does the protocol provide for medical management of potential illness or injury

rovide for safety monitoring? oes the protocol provide for post-e xposure monitoring or fo llow-up? Is it of duration to discover adve rse events which might occur? (g) How and by whom will medical care for res earch-related injuries to subjects be paid

5.

(a) ould accrue to individual subjects?

r om the information likely to be gained through

research? likelihood that each identifi ed societal benefits would be realized?

6.

a) How do the risks to subjects weigh against the anticipated benefits of the research, to ?

7. Ind

(a) Wh proposed research? (b) nve stigators and sponsors of the research? (c) (d) (e) P? (f) ? (g)

8. Informed Consent

a) Will informed consent be obtained from each prospective subject? (b) doc umented, consistent with the requirements of

(c) g ade ation of the risks and di scomforts to subjects from participation in ject or others, and the ri ght to withdraw from the research? i (b) How do propo

(c) W (d) Ho to subjects? (e) How does the protocol p (f) How d long enough

for?

Benefits

What benefits of the proposed researc h, if any, w (b) What benefits to society are anticipated f the research? (c) How would societal bene fits be distributed? Who would benefit from the proposed

(d) What is the

Risk/Benefit Balance

( subjects or to society

ependent Ethics Review

at Institutional Review Board (IRB) reviewed the Is this IRB independent of the i Is this IRB registered with OHRP? Is this IRB accredited? If so, by whom? Does this IRB hold a Federal-Wide Assurance from OHR Are complete records of the IRB review as required by 40 CFR §26.1125 provided What standard(s) of ethical conduct would govern the work?

( Will informed consent be appropriately 40 CFR §26.1117? Do the informed consent materials meet the requirements of 40 CFR §26.1116, includin quate characteriz the research, the potential benefits to the subAppendix B: Framework for Science and Ethics Reviews of Proposed Human Research

38 nglish or other languages among the intended research subjects? (e) investigators and subjects? mforts? (g) ed to inform prospective subjects and to seek and

(h) Wh o ensure fully voluntary participation and to avoid coercion or undue influence?

. Respect for Subjects

) How will information about prospective and enrolled subjects be managed to ensure their

(b) draw from the research at any

(c) How will subjects who decline to participate or who withdraw from the research be dealt (d) What is the literacy rate in E

What measures are proposed to overc ome language differences, if any, between

(f) What measures are proposed to ensure s ubject comprehension of risks and disco What specific procedure will be follow obtain their consent? at measures are proposed t

9

(a privacy? How will subjects be informed of their freedom to with time without penalty?

with?Appendix C: Checklist of El ements Required by 40 CFR §26.1303

39 mission of Completed Human Research for EPA Review

Any person w o s the time of submission information conce ed to EPA, such inform § 26.1303 Sub

h ubmits to EPA data derived from human research covered by this su bpart shall provide at rning the ethical conduct of such research. To t he extent available to the submitter and not previously provid ati on should include:

Requirement Y/N Comments/Page References §1115(a)(

• all re s reviewed,

• s

• a

• p subje 1): Copies of search proposal cientific evaluations, if any, that accompany the proposals, pproved sample consent documents, rogress reports submitted by investi gators, and reports of injuries to cts. §1115(a)(

• atten at the meetings;

• actions taken by the IRB; on these actions including the number of rs voting for, against, and abstaining;

• the basis for requiring changes in or disapproving research;

• 2): Minutes of IRB meetings which shall be in sufficient detail to show dance

• the vote

• membe

a written summary of the discussion of controverted issues and their resolution. §111 5(a)(3): Records of continuing review activities. §1115(a)( orrespondence between the IRB and the investigators. 4): Copies of all c §111

•

to IRB deliberations; relationship between each member and the ll-time employee, a member of governing panel or board, stockholder, paid or unpaid consultant. 5(a)(5): A list of IRB members identified by name; earned degrees; representative capacity; indications of experience su ch as board certifications, licenses, etc., sufficient to describe each mem ber’s chief anticipated contributions

• any employment or other institution, for example, fu

§111 26.11 5(a)(6): Written procedures for the IRB in the same detail as described in § 08(a) and § 26.1108(b).(a) Copies of all of the records relevant to the research specified by§26.1115(a) to be prepared and maintained by an IRB

§1115(a)( new findings provided to subjects, as requ 7): Statements of significant ired by § 26.1116(b)(5). (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 ex pected 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§1125(a)A discussion of

earch.:

(5) The balance of risks and benefits of the proposed res §112 origin 5(b): All information for subjects and written informed consent agreements as ally provided to the IRB, and as approved by the IRB. §112 adve 5(c): Information about how subjects will be recruited, including any rtisements proposed to be used. §112 prese ntial human subjects for the purpose of obtaining t 5(d): A description of the circumstances and methods proposed for nting information to pote heir informed consent. §1125(e): All correspondence between the IRB and the investigators or sponsors.(b) Copies of all of the records relevant tothe information identified in §26.1125(a)-(f)

§ 12 requi revie

1 5(f): Official notification to the sponsor or investi gator, in accordance with the rements of this subpart, that res earch involving human subjects has been wed and approved by an IRB. (c) Copies of sam ment informed consent as specified by §26.1117, bu ple records used to docu t not identifying any subjects of the research (d) If any of th provided, the pers e information lis ted in paragraphs (a) through (c) of this section is not on shall describe the e fforts made to obtain the information.