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 UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

 WASHINGTON, D.C. 20460 

MEMORANDUM:

OFFICE OF 

CHEMICAL SAFETY AND 

POLLUTION PREVENTION

October 1, 2010

SUBJECT: Science and Ethics Review of Protocol for Human Study of Mosquito Repellent 

Performance 

FROM: Clara Fuentes, Ph.D., Efficacy Reviewer 

 Biopesticides and Pollution Prevention Division 

 Office of Pesticide Programs 

Kelly Sherman, Human Research Ethics Reviewer 

 Office of the Director 

 Office of Pesticide Programs 

TO: Linda Hollis, Chief, Biochemicals Pesticide Branch 

 Biopesticides and Pollution Prevention Division 

 Office of Pesticide Programs 

REF: Carroll, S. (2010) Field Efficacy Test of PMD and Lemongrass Oil-Based 

Repellent ‘No Mas’ Against Mosquitoes. Efficacy Test Protocol No Mas 003. 

Unpublished document prepared by Carroll-Loye Biological Research. 340 p. 

We have reviewed the referenced protocol for a field test of mosquito repellency from 

both scientific and ethics perspectives. This review assesses the scientific aspects of the 

proposed research in terms of the recommendations of the final test guideline for Product 

Performance of Skin-applied Insect Repellents of Insect and Other Arthropods Test Guidelines 

(OPPTS Test Guideline No. 810.3700) and of the EPA Human Studies Review Board, and the 

ethical aspects of the proposed research in terms of the standards defined by 40 CFR 26 subparts 

K and L and the recommendations of the EPA Human Studies Review Board. 
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A. Completeness of Protocol Submission 

The submitted protocol was reviewed for completeness against the required elements 

listed in 40 CFR §26.1125. EPA’s checklist is appended to this review as Attachment 5. IRB 

procedures are on file at EPA, and need not be resubmitted. All required elements are present. 

In addition to the protocol itself (pp. 4-32) and the associated consent documents as 

approved by the Independent Investigational Review Board, Inc. (IIRB) (pp. 33-62), the 

following supporting documents were considered in this review: 

• IIRB Approval letter of 7/20/10 (pp. 335-336) 

• Data recording forms (pp. 63-76) 

• Subject training materials for mosquito handling (pp. 77-79) 

• Draft label for product and MSDSs for active ingredients (pp. 81-94) 

• HSR Training records for investigators (pp. 182-183) 

• Index of CLBR-IIRB Correspondence (pp. 194-195) 

• CLBR-IIRB Correspondence (pp. 196-340) 

B. Summary Assessment of Ethical Aspects of the Proposed Research 

Here is a summary of our observations about the ethical aspects of the proposed protocol. 

Supporting details are in the attachment. 

1. Societal Value of Proposed Research: This study will test the efficacy against 

mosquitoes of a lotion formulation containing 16% para-menthane-3,8-diol (PMD) and 

2% lemongrass oil, delivered from a squeeze bottle. EPA requires efficacy testing of this 

specific formulation to support U.S. registration of the product. Direct testing of the 

duration of efficacy is important because consumers, who rely on repellents to avoid 

insect bites, cannot readily assess the efficacy of a product independent of EPA’s 

approval.

2. Subject Selection: Subjects will be recruited from Carroll-Loye Biological Research’s 

(CLBR’s) “Volunteer Database” of previous subjects and others who asked to be added 

to the database. The database is racially diverse, 75% in the age range from 20-40 and 

25% in the range 40-55. The youth and high education levels of candidates in the 

database reflect the university community where the laboratory is located. Explicit 

factors exclude as subjects children, pregnant or lactating women, those in poor health or 

physical condition, and those unable to speak and read English. The sample will thus not 

be fully representative of the population of potential repellent users. Since students and 

employees of the study director and employees of the sponsor will be excluded as 

possible subjects, there are appropriate protections in place to ensure that none of the 

subjects are likely to be subject to coercion or undue influence over their decision to 

participate in the research. 
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3. Risks to Subjects: The protocol and consent forms discuss risks of five kinds: risks 

from exposure to the test material; risks of exposure to biting arthropods; risks from 

exposure to disease vectors; risks of physical stress in the test environment; and risks of 

stress from learning the results of a pregnancy test. All practical steps to minimize 

subject risks have been taken. 

Based on the acute toxicity testing submitted and reviewed by EPA, the test material is 

accurately characterized in the informed consent form as a moderate eye irritant and 

possible skin irritant. 

Because of the generally low acute and chronic hazard profile of the test material, the 

design of the research to minimize exposure, and the training of subjects to remove 

mosquitoes before they bite, the probability of occurrence of the identified hazards is 

accurately characterized as “extremely small.” 

4. Benefits: There are no direct benefits to subjects. This is made clear in the protocol and 

informed consent form. If the testing shows good efficacy, the direct beneficiary of the 

research is likely to be the sponsor. 

5. Risk/Benefit Balance: No practical opportunities to further reduce risk to subjects 

while maintaining the robustness of the scientific design have been overlooked. The 

residual risk to subjects is very low, and reasonable in light of the potential benefits of the 

data to future repellent users. 

6. Independent Ethics Review: The Independent Investigational Review Board, Inc. 

(IIRB), of Plantation, Florida, has reviewed and approved the protocol and informed 

consent materials. IIRB is independent of the investigators and sponsors. Satisfactory 

documentation of IIRB procedures and membership is on file with the Agency. 

7. Informed Consent: The protocol contains a complete and satisfactory description of the 

process by which potential subjects will be recruited and informed and for seeking their 

consent to participate. A copy of the IRB-approved consent documents meeting all 

requirements of 40 CFR §§26.1116 and 26.1117 is included in the proposal. 

8. Respect for Subjects: Methods proposed for managing information about prospective 

and enrolled subjects are adequate to protect their privacy. Subject names and other 

personal information are linked on only one form to their arbitrary “subject number”; in 

all other data collection forms subjects are identified only by their assigned number. 

Subjects will be free to withdraw at any time, and will be reminded of this at several 

points before and during the research. Subjects who withdraw will be compensated for 

time spent up to the point of withdrawal. Medical care for research-related injuries will 

be provided at no cost to the subjects. 
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C. Compliance with Applicable Ethical Standards 

This is a protocol for third-party research involving intentional exposure of human 

subjects to a pesticide, with the intention of submitting the resulting data to EPA under the 

pesticide laws. Thus the primary ethical standards applicable to this proposal are 40 CFR 26, 

Subparts K and L. In addition, the requirements of FIFRA §12(a)(2)(P) for fully informed, fully 

voluntary consent of subjects apply. Because the test will be conducted in California, the 

provisions of the California Code of Regulations, Title 3, §6710 apply as well, including 

provision to subjects of the “Experimental Subject’s Bill of Rights” appearing on p. 38. 

A point-by-point evaluation of how this protocol addresses the requirements of 40 CFR 26 

Subparts K and L and the criteria recommended by the HSRB is appended as Attachment 1. 

The protocol should be revised before study execution to exclude as permissible subjects 

employees of the study sponsor. 

40 CFR 26 Subpart L, at §26.1703, as amended effective August 22, 2006, provides in 

pertinent part: 

EPA shall not rely on data from any research involving intentional exposure of 

any human subject who is a pregnant woman (and therefore her fetus), a nursing 

woman, or a child. 

This protocol requires that subjects be at least 18 years old and excludes female subjects who are 

pregnant or lactating. Thus §26.1703 would not forbid EPA to rely on a study executed 

according to this protocol. 

D. Summary Assessment of Scientific Aspects of the Proposed Research 

The study will test the efficacy under field conditions of a new mosquito repellent lotion 

containing 16% plant derived p-menthane-3,8-diol (PMD) and 2% lemongrass oil (citral). The 

main objective of the study is to quantify the efficacy of the formulation to repel mosquitoes in 

the field. The protocol’s objective is to determine the duration of efficacy of the lotion 

formulation in repelling three mosquito species (Culex, Anopheles, and Aedes) when applied at a 

typical consumer dose. The endpoint will be the time of repellent failure expressed as the time 

of First Confirmed Landing with Intent to Bite (FCLIBe) for each subject. Efficacy is expressed 

for each subject as Complete Protection Time (CPT), which is the period from application of the 

repellent until time of FCLIBe, or conclusion of the test, whichever occurs first. Mosquito 

specimens will be collected from treated and untreated subjects and taken to the laboratory for 

identification, using taxonomic keys and stereomicroscopy. Disease vectors will be screened for 

transmissible viruses, and any positive results will be provided to the appropriate test subject(s). 
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1. Study design: 

The duration of efficacy of the test material will be evaluated in the field with ten treated 

subjects, five of each sex, at each of two sites representing distinct ecological habitats. Three 

additional subjects will be enrolled as alternates to replace any subjects who may withdraw 

prematurely from the test, or who may fail to show up for testing. Untreated subjects will 

monitor activity at the test sites throughout the duration of the test by exposing either their lower 

leg or forearm for periods of 1 minute every 15 minutes. Minimum acceptable ambient biting 

pressure is defined as 1 landing with intent to bite (LIBe) per minute. Untreated subjects will not 

be compared to treated subjects. 

2. Statistical design: 

The aim of the research is to characterize the duration of repellency defined as Complete 

Protection Time (CPT). The mean of ten individual subject values will be used to estimate a 

central measurement of CPT at each test site. The protocol characterizes ten subjects as a 

compromise between ethical and economic considerations, based on the rationale that ten 

subjects are two-thirds more than the historical EPA requirement of six subjects. Furthermore, if 

few values are censored, and particularly, if the range of values is not great, a sample size of ten 

should give excellent estimates of mean, median, and variation around those values, relative to 

historical standards. 

While EPA has recently accepted repellent efficacy data based on a sample size of ten, 

EPA recommends that the investigator revise and expand the explanation offered for the 

proposed sample size. EPA thinks that reference to historical standards is irrelevant in light of 

current guidelines and should be deleted. EPA’s guidelines (OPPTS Test Guideline No. 

810.3700) provide: “(v) Sample size. The sample should be large enough to be likely to yield a 

definitive answer to the research question being addressed, and its size should be justified 

statistically in each protocol, taking into account the specific characteristics of the proposed 

research and the desired accuracy and precision of the results. Researchers are encouraged to 

consult a statistician to help determine appropriate sample size.” Therefore, investigator should 

revise the rationale in the protocol to include a statistical justification for the proposed sample 

size in light of the desired accuracy and precision of the results. 

3. How and to what will human subjects be exposed? 

Subjects will be exposed to test material and mosquitoes in the field. Each subject will 

be treated on one forearm or lower leg, depending of feeding behavior of predominant mosquito 

species active in the field. A standardized typical dose, expressed as volume per unit area, will 

be scaled to the measured surface area of each subject’s forearm or lower leg, and applied by 

technicians using tuberculin (1 ml) syringes. The test material has been tested in animals for 

acute toxicity. The NOAEL for acute dermal LD50 of No Mas is greater than 5,000 mg/kg body 

weight. Thus, exposure of 14.3 mg/kg of lotion, containing 18.0% of combined active 

ingredients, results in an estimated MOE > 100. Most inert ingredients in the formulation are 

classified by the Agency as relative safe for all uses and have a low acute and chronic risk 
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profile. Only a few ingredients present ocular and dermal irritation potential; other ingredients 

are commonly used in drugs and cosmetic products. Subjects with known allergic reactions or 

other skin conditions are excluded from participation in the test. Data from animals studies show 

that the formulation is of low toxicity by oral and dermal routes of exposure. It is not irritating to 

the skin, and it is not a skin sensitizer. 

Results from toxicity testing: 

• Primary eye irritation study on rabbits shows that the No Mas is a moderate 

irritant to the eyes. 

• Dermal sensitization study in Guinea pigs (Buehler method) shows that the test 

material is not a contact sensitizer. 

• Primary skin irritation in rabbits study shows that No Mas is moderately irritating 

to the skin. 

• The single dose acute dermal LD50 of the formulation is >5,000 mg/kg in male 

and female rats. 

• The acute oral LD50 of No Mas is >5,000 mg/kg in male and female rats 

Subjects will be exposed to naturally occurring populations of mosquitoes in field sites 

where tests are being conducted and mosquito-borne pathogens have not been detected by PCRbased screening of trapped mosquitoes for at least 2 weeks. Subjects with known allergic 

reactions to mosquito bites will be excluded from research participation. 

4. Endpoints and Measures: 

A standard “typical consumer dose” will be determined in the dose determination phase, 

and used for each subject in the repellency phase. The end point of the dose determination phase 

will be a standard volumetric rate of application expressed in ml/cm2

. 

In the repellency phase, efficacy will be measured as Complete Protection Time (CPT) 

for each subject, defined as the time between application of test material and the First Confirmed 

Landing with Intent to Bite (FCLIBe). A Landing with Intent to Bite (LIBe) occurs when a 

mosquito alights on treated skin and extends its proboscis to the skin while ceasing locomotion. 

First Confirmed LIBe (FCLIBe) is defined as a LIBe followed by another LIBe within 30 

minutes. The end point will be the time of repellent failure expressed as the time of the FCLIBe 

for each subject. Repellency will be expressed as average CPT, which is measured as the mean 

time across subjects from initial application of a typical consumer dose to the time of FCLIBe. 
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E. Compliance with Applicable Scientific Standards 

This protocol adequately addresses the following elements according to applicable 

scientific standards: 

• Prerequisite acute toxicity research to characterize toxicological profile of the 

formulation and calculate margin of exposure (MOE). 

• Dosimetry 

• Experimental design 

• Pre-training of subjects on how to handle mosquitoes. 

The following elements in the protocol require revision before the research goes forward: 

• The statistical analysis plan should discuss how non-normally distributed data points will 

be treated. That is, how the researcher plans to analyze and interpret results from nonnormally distributed data points that may follow an unknown distribution. 

• On page 28, line 41, delete the phrase “for each test material” because this protocol is for 

testing one single formulation. 

• The sequential exposure table on page 74 should include 80 sequential exposure 

intervals. It is not clear why only 36 counts are tabulated, and how the 44 remaining 

intervals can be accommodate one single column, labeled as “etc.” If the test is going to 

last more than 9 hours, a second page will be needed. 

 Attachments: 

1. Summary Review of Protocol NO-MAS 003 dated 7/15/2010 

2. §26.1111 Criteria for IRB approval of research 

3. §26.1116 General requirements for informed consent 

4. §26.1117 Documentation of informed consent 

5. §26.1125 Criteria for Completeness of Proposals for Human Research
Attachment 1 

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EPA Protocol Review: NO-MAS 003 

Title: Field Efficacy Test of PMD and Lemongrass Oil-Based Repellent ‘No Mas’ 

Against Mosquitoes. 

Date: July 15, 2010 

Principal Investigator and any sub-investigators: 

 Scott P. Carroll, Ph.D. 

 

Participating Laboratories: 

 Carroll-Loye Biological Research, Inc. 

 711 Oak Avenue 

 Davis, CA 95616 

Sponsor: Sam Darling 

415 Wilkie Way 

Salt Spring Island 

British Columbia V8K2J4 Canada 

IRB: Independent Investigational Review Board 

 6738 West Sunrise Blvd. Suite 102 

 Plantation, FL 33313 

1. Societal Value of Proposed Research 

(a) What is the stated purpose of the proposed research? 

The objective of the dosimetry phase “is to determine the amount of repellent a consumer 

might typically apply using each of the delivery systems in the study, and to determine 

the dosage to be applied during the repellency phase of the study. The endpoint will be a 

standard rate of application expressed in ml/cm2

.” (p. 4) 

The objective of the repellency phase is to determine the duration and efficacy of the test 

Material(s) in repelling three species of mosquitoes (Culex, Anopheles, and Aedes) when 

applied at a typical consumer dose. (p. 4) 

“Efficacy and duration will be measured as Complete Protection Time, or CPT, defined 

herein as the time between application of test material and the First Confirmed Probing 

called ‘Landing with Intent to Bite’, or ‘LIBe’, which is defined as when a mosquito 

alights on the treated test skin of a subject and extends its proboscis to the skin surface 
Attachment 1 

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while ceasing locomotion. A ‘First Confirmed LIBe’ is that which is followed by another 

within 30 minutes.” (p. 4) 

(b) What research question does it address? Why is this question important? 

Would the research fill an important gap in understanding? 

“The US Centers for Disease Control (CDC) has acknowledged the existence of 

substantial consumer interest in new and effective insect repellent products, including 

the choice of a variety of formulations, delivery systems, and concentrations of active 

ingredient. US EPA has requested new, US-based efficacy data as a condition of 

registration for the test material(s). The rationale for this study is to provide those 

efficacy data. The information will also be used in product labeling and for increased 

user acceptance.” (p. 5) 

(c) How would the study be used by EPA? 

EPA will consider the study in defining acceptable label claims for repellent efficacy for 

the test material. 

(d) Could the research question be answered with existing data? If so, how? If not, 

why not? 

EPA requires product-specific efficacy data to support its registration. No previous 

testing of this product against mosquitoes under proposed used pattern has been 

conducted. 

(e) Could the question be answered without newly exposing human subjects? If so, 

how? If not, why not? 

“Human subjects are required because they represent the target system for the test 

material, and sufficiently reliable models for repellency testing have not been 

developed.” (page 5: line 14 - 16). 

2. Study Design 

(a) What is the scientific objective of the study? If there is an explicit hypothesis, what 

is it? 

The main objective of the study is to quantify the lasting efficacy of the formulation to 

repel mosquitoes in the field. The specific protocol’s objective is to determine the 

duration of efficacy of the lotion formulation in repelling three mosquito species (Culex, 

Anopheles, and Aedes) when applied at a typical consumer dose under field conditions. 
Attachment 1 

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(b) Can the study as proposed achieve that objective or test this hypothesis? 

 The objective cited may be achieved by the study as proposed if the protocol is revised 

 and amended to explain, in more detail, the following: 

• How will these data be summarized, analyzed and interpreted if the results show that 

the data points are not normally distributed and the data follows an unknown 

distribution? 

• What measurements will be taken to minimize right censoring? 

• How does the researcher plan to minimize, and account for variability in the 

experiment? 

2.1 Statistical Design 

(a) What is the rationale for the choice of sample size? 

The rationale for this sample size appears on pp. 18 to 20 (§ 4.1) of the study protocol. 

Researcher’s justification for sample size is based on the argument that a 10 subjects 

sample reflects a compromise between cost and precision; ten subjects are two-thirds 

more than the historical EPA requirement of 6 subjects. In Addition, it is stated in the 

study protocol, page 19: lines 30 -33, that “If a minority of values is censored, and 

particularly if the range of values is not great, a sample size of 10 should give an 

excellent estimates of mean, median, and variation around those values, relative to 

historical standards.” 

(b) What negative and positive controls are proposed? Are proposed controls 

appropriate for the study design and statistical analysis plan? 

There are no comparison materials or positive controls. Omission of matrix and 

comparison materials is appropriate for the study design. No direct comparisons of 

treated and untreated subjects are contemplated in the statistical analysis plan. 

(c) How is the study blinded? 

n/a. There is one treatment only. 

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

All subjects will be assigned to the treated group, which will be stratified by gender. 

Table 8 on page 24 summarizes the proposed distribution of test material among subjects. 

The treatments will be balanced between arms and legs if both limbs are used, depending 

on the feeding behavior of mosquitoes present in the field. Two untreated subjects are 

also shown in table 8. Their untreated skin will be balanced between arms and legs 

similarly to treated subjects. 
Attachment 1 

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(e) Can the data be statistically analyzed? 

Yes. The application rate for efficacy testing will be an average typical consumer dose 

estimated from dosimetry analysis (§ 4.6) developed by CLBR laboratory. Subjects’ 

dose means based on 3 applications per subject will be used to calculate dose grand mean 

(± SD) across all 10 subjects. In the efficacy phase, 10 individual subject values for CPT 

will be obtained and analyzed. CPT will be measured as a single time value per subject. 

Mean CPT will be calculated across all 10 subjects, with standard deviation and 95% 

confidence interval (page 29: lines 22 to 27). Kaplan-Meier survival analysis will be 

employed to accommodate some data censoring. The Kaplan-Meier test will be also 

employed to estimate the median CPT, and the time until 25% efficacy failure of the test 

material. In the presence of high censoring frequency, median and mean values will be 

underestimated. Ambient landing pressure will be measured as number of landings per 

untreated subject per exposure period and span of exposure (page 29). 

(f) What is the plan for statistical analysis of the data? 

“Statistics will be computed with SAS’s JMP software, Version 5.0.1.2 (SAS 

Institute, Cary, NC).” (p.28) 

“In the efficacy phase, CPT, defined as period between application of formulation and 

FCLIBe or end of test, will be measured as a single time value for each subject. A mean 

CPT will be calculated across all 10 subjects, and will be presented with standard 

deviation and 95% confidence interval. Data will be normalized as possible to enhance 

the value of confidence interval calculations.” (p. 29). 

 “To examine the temporal pattern of failure further, we will employ Kaplan-Meier 

survival analysis by subject. Kaplan-Meier survival analysis accommodates some 

data censoring in the event that any subjects withdraw or are withdrawn before 

failure. In addition, we will estimate the Kaplan-Meier median, and the time until 

25% failure, for each test product. In the presence of a high frequency of censoring, 

median (and mean) values will be underestimated.” (p.29) 

(g) Are proposed statistical methods appropriate to answer the research 

question? 

The proposed statistical methods are based on assumption of normality, low frequency of 

right censoring, and historical standards. The researcher should elaborate on the 

appropriate methodology to meet such assumptions, and provide scientifically- based 

rationale for anticipating that those assumptions will be met. 

 

(h) Does the proposed design have adequate statistical power to definitively 

answer the research question? 

The proposed research will produce a data set more robust than most on which past 

decisions by EPA concerning acceptable claims of repellency have been based. An 
Attachment 1 

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increase in sample size from six to ten subjects will improve precision and is consistent 

with suggestions from HSRB to EPA (pp. 29 – 30). Historical standards are irrelevant in 

light of EPA current guidelines (OPPTS Test Guideline No. 810.3700). A sample size of 

10 may be adequate depending on experimental variability, experimental design and 

interpretation of test results. 

2.2 How and to what will human subjects be exposed? 

Subjects will be exposed to test material and mosquitoes in the field. A standardized 

typical dose, expressed as volume per unit area, will be scaled to the measured surface 

area of each subject’s forearm or lower leg and applied by technicians using tuberculin (1 

ml) syringes. The repellent’s active ingredients have a low acute and chronic risk profile 

(§2) (page 5: lines 40 to page 5: lines 1 to 21), and the inert ingredients are classified by 

the Agency as safe for use. Only a few ingredients of the inert ingredients present ocular 

and dermal irritation potential. The other ingredients are commonly used in drugs and 

cosmetic products. The test material has been tested in animals for acute toxicity. 

Subjects with known allergic reactions (§3.3.3 and §3.3.5), or other skin conditions 

(§3.3.4) are excluded from participation in the test. 

Subjects will be exposed to naturally occurring populations of mosquitoes in field sites 

where tests are being conducted and mosquito borne pathogens have not been detected 

for at least 2 weeks (§1.3.2; page 7: lines 23 to page 8: 10). Subjects with known 

allergic reactions to mosquito bites will be excluded from research participation ((§3.3.3). 

(a) What is the rationale for the choice of test material and formulation? 

Efficacy data to support label claims for this product are required by EPA as a condition 

of registration. EPA requires submission of efficacy data for all products claiming 

efficacy against human health pests. 

(b) What is the rationale for the choice of dose/exposure levels and the staging of 

dose administration? 

Dosimetry is the procedure employed in this study to estimate the standard consumer 

dose used for testing efficacy. 

(c) What duration of exposure is proposed? 

The repellency phase will last from 12 to 18 hours; the period of actual exposure is 

uncertain; it may be less than 8 hours (excluding travel time) to as long as 18 hours, 

depending on how long the repellent remains effective (page 3 in Informed Consent 

Authorization to Participate as a Research Study Subjects). Data sheets for Mosquito 

LIBes at 15 Minutes Intervals show 80 data points, which corresponds to 20 hours of 

observations. Although not specified, the reviewer assumes that the total hours of 

observations will be equally divided between the 2 testing sites, and it will be 10 hours of 

total observations per site. That assumption needs to be verified by the researcher. 
Attachment 1 

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2.3 Endpoints and Measures 

(a) What endpoints will be measured? Are they appropriate to the question(s) being 

asked? 

“Subject measurements” include subjects’ limbs surface area, which may be measured or 

based on historical subject data as explained. “This data will be kept on file for each 

subject. Subjects will be re-measured biennially or if, when asked, they indicate they may 

have gained or lost weight or muscle mass on their limbs since their measurements were 

last taken.” (page 21: lines 1 – 10). Volume of test material delivered to the skin is 

described in §4.7. (p. 23). These measurements are appropriate to estimate the standard 

consumer dose employed for efficacy evaluation. 

Endpoints for efficacy evaluation (page 29: lines 8 – 14): 

• Exposure delay (min)-time between application and first exposure 

• Minutes to FCLIBe. 

• Complete Protection Time (CPT)–time between application and FCLIBe or end of 

test. 

• Total number of landing mosquitoes per minute on untreated subjects at the beginning 

of each interval. 

Data forms are presented for dosimetry (limbs’ measurements on page 64; grams of 

applied product on page 65; randomized treatment allocation on page 66, and repellent 

applications on page 67); Field Environmental conditions on page 68; Researcher Notes 

on page 69, and Mosquito LIBes at 15 minutes intervals on pages 70 – 73 (these data 

sheets include 80 exposures of one minute each at 15 minutes intervals per subject); one 

data sheet for 36 Sequential Exposure Intervals up to “etc” on page 74; Research Subject 

Tracking form on page 76, and data sheet for Pools of Mosquito Species submitted to 

UCD by CLBR on page 75. The sequential exposure table on page 74 should include 80 

sequential exposure intervals. It is not clear why it stops at 36 counts, and how the 44 

remaining intervals would be accommodated under one column labeled as “etc.” 

(b) What steps are proposed to ensure measurements are accurate and reliable? 

• Test material will be applied by laboratory technicians. 

• Alternate subjects will be enrolled to ensure adequate sample size. 

• Subjects will be trained to handle mosquitoes and to remove them before they can 

bite. 

• All landings are verified and recorded by a research technician. 

(c) What QA methods are proposed? 

“A separate, professional Quality Assurance Unit (QAU) will inspect the study. The 
Attachment 1 

Page 14 of 30 

QAU will report to the Study Director of Operations. Protocol Review and Comments 

must take place before data collection commences. In-Life Inspection must include 

observing the measurement and recording of key variables by subjects and 

technicians. In addition, the Final Report will be audited for completeness and 

accuracy. A QAU Statement will address compliance and noncompliance or any 

omissions in auditing. Findings from the In-Life Inspection and the Final Report, as 

well as the QUA statement will be transmitted to both, the study director and to the 

sponsor Monitor.” (p. 30). 

(d) How will uncertainty be addressed? Will point estimates be accompanied by 

measures of uncertainty? 

“Mean CPT will be calculated across all 10 subjects per treatment, and will be 

presented with standard deviation and 95% confidence interval.” (p.29) 

3. Subject Selection 

3.1 Representativeness of Sample 

(a) What is the population of concern? How was it identified? 

The population of ultimate concern consists of people who would purchase and use 

mosquito repellents. Little information is available to characterize this population, 

but it is presumed that repellent users are highly diverse in age, gender, physical size, 

general health, attractiveness to mosquitoes, and other characteristics. The population 

from which subjects are recruited appears to be chosen largely on the basis of 

convenience, and is not screened for past or likely future use of repellents. 

 (b) From what populations will subjects be recruited? 

“For reasons of practicality and control, we work with people associated with the 

community in which our business is located (Davis, CA). Davis is a university 

dominated community, and so the population demography differs somewhat from 

non-university communities. Compared to the Population of Concern (the US 

population - all potential repellent users), our sampling frame tends to under-represent 

blacks and over-represent Asians. It is also young, well educated, and slanted 

towards life science researchers and students. 

“Over time, we have developed a Volunteer Database of individuals who have 

expressed interest in participating in future repellency tests, provided contact 

information, and asked us to contact them. Initial recruiting is from this database, 

then from word-of-mouth of volunteers. The size and composition of the database 

varies over time as new individuals volunteer and old volunteers move out of the 

Davis area, but is now typically over 100 individuals, with the following average 

ethnic (self-identified) and gender distribution (averaged over 3 years): 
Attachment 1 

Page 15 of 30 

Male 52% 

Female 48% 

Caucasian 73% [72% on IIRB Site Questionnaire Form]

Asian 12% 

Hispanic 8% 

African-American 3% 

Middle Eastern 5% 

“In general, about three-quarters of the subjects are age 20-40, with the remainder 

between 40 and 55. Final composition is not determined until enrollment is completed. 

The relevant demographics of the participants will be reported.” (p. 12)

 (c) Are expected participants representative of the population of concern? 

 If not, why not? 

“Based on review of the scientific literature regarding individual differences in 

repellent performance and attractiveness to mosquitoes, we conclude that this study’s 

deviations from the ideal frame will not influence the representativeness of the 

results, or their generalizability to the greater population. Lastly, because our 

Volunteer Database cohort is comprised by individuals who regularly spend time in 

outdoor setting (and thereby may have relatively frequent encounters with biting 

arthropods), this group is probably appropriate for insect repellent users in general. ”

(pp. 13-14) 

By excluding children, pregnant or lactating women, non-English speakers, and those 

in poor physical condition, among others, the exclusion criteria will mean that 

participants will not be representative of at least some segments of the population of 

concern. 

 (d) Can the findings from the proposed study be generalized beyond the study sample? 

Yes, because the study will be replicated twice at 2 different mosquitoes habitats. 

3.2 Equitable Selection of Subjects 

(a) What are the inclusion/exclusion criteria? Are they complete and appropriate?

“Inclusion: 

1. Age 18-55; 

2. Written consent; and 

3. Speak and read English. 
Attachment 1 

Page 16 of 30 

Exclusion: 

1. Known to be hypersensitive to mosquito bites; 

2. Phobic of biting insects or insect bites; 

3. Known to be allergic to insect repellents or common cosmetics; 

4. Known to be sensitive to any of the test material ingredients; 

5. Poor physical condition; 

 

6. Unwilling to submit to brief query about personal condition; 

7. Use of insect repellent within one day proceeding the efficacy test; 

8. Unwilling to refrain from use of perfumed products, alcoholic beverages or 

smoking after 9 pm the evening preceding the efficacy test and throughout 

that test; 

9. Known to be pregnant or lactating; 

10. Unable to deliver the test material to own left and right limbs for dosimetry; 

11. Unable to see biting insects on skin or otherwise effectively monitor and 

remove biting insects that contact skin; 

12. Student or employee of Study Director; and 

13. Does not regularly spend time in outdoor settings.” (page 14-15) 

Exclusion factor #12 should be revised to exclude employees of the sponsor, as well as 

students or employees of the study director. 

(b) What, if any, is the relationship between the investigator and the subjects? 

Subjects are recruited from “the community in which [the Investigator’s] business is 

located . . . . Over time, we have developed a Volunteer Database of individuals who 

have expressed interest in participating in future repellency tests, provided contact 

information, and asked us to contact them. Initial recruiting is from this database, 

then from word-of-mouth of volunteers.” (page 12) 

Students and employees of the Study Director are excluded from participation. (page 

15) 
Attachment 1 

Page 17 of 30 

 (c) If any potential subjects are likely to be especially vulnerable to coercion or 

 undue influence, what is the justification for including them? 

None of the subjects are expected to be especially vulnerable to coercion or undue 

influence. 

(d) What process is proposed for recruiting and informing potential subjects? 

The recruiting/informing process to be used is described in the protocol on pp. 14-17. 

(e) If any subjects are potentially subject to coercion or undue influence, what 

specific safeguards are proposed to protect their rights and welfare? 

Students and employees of the Study Director are excluded from participation. (p. 15) 

No eligible subjects are potentially subject to coercion or undue influence. 

3.3 Remuneration of Subjects

(a) What remuneration, if any, is proposed for the subjects? 

“[E]ach research study participant will receive a cash payment of $20 per hour...If 

you are designated as an ‘alternate subject’ you will be paid for the hours you spent 

being trained, plus you will receive a payment of $50 to compensate for being 

inconvenienced.” (page 41) 

(b) Is proposed remuneration so high as to be an undue inducement? 

No. 

(c) Is proposed remuneration so low that it will only be attractive to economically 

disadvantaged subjects? 

No. 

(d) How and when would subjects be paid? 

“Payment will be made at the end of each visit or whenever you withdraw from the 

study.” (page 41) 

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 material? 
Attachment 1 

Page 18 of 30 

Acute toxicity studies required for the registration of an insecticide were conducted 

on animals. Data from animals studies show that the formulation is of low toxicity by 

oral and dermal routes of exposure. It is not irritating to the skin, and it is not a skin 

sensitizer. 

Results from toxicity testing: 

• Primary eye irritation study on rabbits shows that the No Mas is a moderate 

irritant to the eyes. 

• Dermal sensitization study in Guinea pigs (Buehler method) shows that the test 

material is not a contact sensitizer. 

• Primary skin irritation in rabbits study shows that No Mas is moderately irritating 

to the skin. 

• The single dose acute dermal LD50 of the formulation is >5,000 mg/kg in male 

and female rats. 

• The acute oral LD50 of No Mas is >5,000 mg/kg in male and female rats. 

 (b) What is the nature of the risks to subjects of the proposed research? 

The protocol discusses risks of five kinds for subjects participating in the repellency 

phases of the study: risks from exposure to the test material; risks from exposure to 

biting arthropods; risks from exposure to disease vectors; risks of physical stress in 

the test environment; and risks of stress from learning the results of a pregnancy test. 

Each class of risk and the steps taken to minimize it is discussed in pp. 5-10. The 

same classes of risk are characterized in the informed consent documents for treated 

and untreated repellency subjects on pp. 38-40 and pp. 48-50. There are two kinds of 

risks discussed for subjects in the dosimetry phase: risks from exposure to the test 

material; and risks of stress from learning the results of a pregnancy test; these types 

of risks are characterized in the consent documents for dosimetry subjects on p. 58. 

(c) What is the probability of each risk associated with the research? How was this 

probability estimated? 

No numerical probability is estimated. Potential subjects are told (with respect to the 

risks of contracting a disease carried by mosquitoes if they are bitten) that “there is a 

very sight possibility that you will contract a disease carried by mosquitoes if you are 

bitten, such as West Nile virus or equine encephalitis. This test will be conducted in 

an area in which such viruses have not been found in captured mosquitoes for at least 

two weeks. The risk is probably very low that any individual mosquito that might 

bite you carries a disease. In addition, since you are wearing repellent and/or other 

protective measures, and are carefully watching for mosquitoes that land and try to 

bite, you are probably at no more risk than you would experience when engaged in 

normal outdoor activities in a similar rural area at the same time of year.” (page 36) 
Attachment 1 

Page 19 of 30 

4.2 Risk minimization 

(a) What specific steps are proposed to minimize risks to subjects? 

Risks from exposure to the test material. 

• Candidates with known allergic reactions to insect repellents and common 

cosmetics are excluded. 

Risks from Exposure to Biting Arthropods and Risks from Exposure to Disease 

Vectors.

• The risk of a skin reaction to an insect bite is reduced by excluding candidate 

subjects who are aware of having a history of such reaction. 

• Subjects will be trained to operate a mechanical aspirator to remove any biting 

insects that land on skin and how to identify insects that have landed with the 

intent to bite. 

• In the field, treated subjects will expose small areas of treated skin for 4 

minutes per hour. Untreated subjects will be protected with gloves, head nets, 

and Tyvek body suits. They will expose untreated skin for up to 2 minutes 

every half hour. 

• The research is being conducted in an area where insect vectored viruses have 

not been detected by PCR-based screening of trapped mosquitoes from the 

test sites for at least two weeks, so the risk is probably low that any individual 

insect present carries a disease. Approximately 1000 mosquitoes of all 

species combined will be trapped for each sample and samples will be taken 

within two weeks prior to the test date. 

• Of the diseases vectored by mosquitoes that may be present at the field site, 

only West Nile Virus (WNV) is of particular concern. The US Centers for 

Disease Control estimates that about 4 out of 5 people who are infected with 

WNV will not develop any type of illness. Subjects are instructed to be alert 

for any flu-like symptoms (unusual tiredness or unusually severe headaches, 

body aches, fever), glandular swelling or a rash on the trunk of the body, for 

up to two weeks after the test. About 1-in-150 infected people will develop 

more serious symptoms, which are described to the subjects. Most people 

(about 4 out of 5) who are infected with WNV will not develop any type of 

illness. Added risk from Western Equine Encephalitis and St. Louis 

Encephalitis, which present with similar pathology, is extremely low. 

• First Aid materials will be available on-site. 

• Epi-Pens will be on-site to treat anaphylactic allergic reactions. 
Attachment 1 

Page 20 of 30 

• Subjects will be trained in the laboratory to handle mosquitoes using 

aspirators. Subjects will observe mosquitoes’ behavior in the lab, and will 

learn how to remove them before they have time to bite. 

Risks of Physical Stress in the Test Environment.

• A physician who has read the protocol and discussed the research with the 

Study Director will be on call on the day of field testing. 

Risks of Stress from Learning the Results of a Pregnancy Test.

• Results of pregnancy testing will be observed by one female technician only 

and never recorded to minimize the stress on a female subject testing positive, 

and minimize the possibility that other staff or subjects may become aware of 

the results of that test. 

(b) How do proposed dose/exposure levels compare to established NOELs/NOAELs 

for the test material? 

The estimated dose rate is 14.3 mg/kg (Table 5 in page 20) based on predicted mean 

grams of lotion (rounded to 1 g.) applied by subject in prior Carroll-Loye Biological 

Research (CLBR) insect repellent efficacy studies (Table 4 in page 21). 

The NOAEL for acute dermal LD50 of No Mas is greater than 5,000 mg/kg body weight. 

Thus, exposure of 14.3 mg/kg of lotion, containing 18.0% of combined active 

ingredients, results in an estimated MOE > 350. 

 (c) What stopping rules are proposed in the protocol? 

“Stop Rules 

All subjects 

1. Consented duration reached. 

 

2. Test site becomes unsafe for subjects for any reason. 

 

3. Foraging pressure falls below threshold needed to challenge the test 

 material. 

 4. Biting/foraging pressure falls below threshold needed to challenge the 

 test material. 

5. Sustained wind speeds exceeds 10 mph 
Attachment 1 

Page 21 of 30 

Individual subjects 

1. Subject asks to withdraw 

 

2. Subject proves unattractive to target species 

3. Subject’s treated limb receives Confirming LIBe 

4. More than one mosquito attempts to bite the treated subject’s treated 

 limb during any exposure period 

5. Exhibits hypersensitivity to insect bites during test 

6. Exhibits sensitivity to the test material during the test 

7. Medical management is invoked for the subject (§1.3.6)” (pp. 25-26) 

(d) How does the protocol provide for medical management of potential illness or 

injury to subjects? 

“If you are injured as a result of being in this study, a consulting physician who is 

aware of the study will be contacted immediately by telephone. Medical treatment 

will be available from a health care facility.” (p. 40) 

(e) How does the protocol provide for safety monitoring? 

“Subjects are clearly and repeatedly informed that they may remove themselves for 

any reason from the study at any time, without penalty to their compensation. All 

subjects are asked to contact the Study Director and a physician of their own choice at 

any time should they develop a rash (a delayed hypersensitivity reaction) within 7 

days of the conclusion of the test day. 

“On the day of any study visit, staff will immediately communicate all subject 

concerns about health, safety, or comfort to the Study Director for assessment. The 

Study Director will also assess skin condition of affected subjects should any bites 

inadvertently occur during efficacy testing, or any subject reports any discomfort in 

treated areas. Subjects are instructed to inform the Study Director (i.e., the ‘Principal 

Investigator’), or any other staff member if at any time during the study a subject 

suffers a skin reaction, such as redness, edema, itching or pain, or feels ill. Such 

subjects will be immediately withdrawn from testing and insect exposure, and 

medical management will be implemented. When a subject completes the study or is 

removed for any reason, treated skin areas will be gently washed with clean water and 

mild soap, rinsed with a 35% ethanol in water solution, then gently dried with a towel 

to remove test material. 

 
Attachment 1 

Page 22 of 30 

“When medical management is implemented, the Study Director will contact the OnCall physician for the study and comply with the physician’s instructions. On the day 

of testing, a physician who has read the protocol and discussed the research with the 

Study Director will be on call. Contact information for the nearest medical facilities 

and maps from the test site to the facilities will be prepared and on file before the day 

of testing. In unlikely event of a Type 1 allergic reaction (anaphylaxis), we will 

contact 9-1-1 by cellular or satellite telephone and cooperate as instructed with 

emergency personnel. Epi-Pens will be on-site. At least one qualified researcher will 

remain with the other test subjects if other researchers depart with an injured or ill 

subject. We will be prepared to instruct emergency personnel on how to reach our site 

via multiple routes. In addition, we will personally transport affected persons to the 

nearest hospital if so advised by emergency personnel. There is sufficient redundancy 

in personnel that in such a case subjects remaining at the study site will still receive 

appropriate technical, scientific and safety guidance. 

“Subjects may also request access to standard first aid materials (such as bandages, 

antiseptics, and mild topical and oral antihistamines) and request qualified first aid 

assistance at any time. 

“As part of Medical Management, the Study Director will record all benign and 

adverse health observations.” (pp. 8-9) 

(f) How does the protocol provide for post-exposure monitoring or follow-up? Is it 

of long enough duration to discover adverse events which might occur? 

“Contact a physician and the Principal Investigator if you develop a rash within 7 days 

after the day of testing.” (page 38) Irritant or allergic reactions to the test material or 

to mosquito bites are likely to occur shortly after exposure. Therefore, the seven-day 

period provides long enough duration to discover adverse events 

(g) How and by whom will medical care for research-related injuries to subjects be 

paid for? 

“Carroll-Loye Biological Research will cover the costs of such medical treatment that 

are not covered by your own insurance or the insurance of a third party under which 

you are covered. If necessary, Carroll-Loye Biological Research will transport you to 

receive medical attention and pay costs associated with the reasonable and 

appropriate treatment for any injuries incurred as a direct result of participation in the 

study.” (page. 40) 

5. Benefits 

(a) What benefits of the proposed research, if any, would accrue to individual subjects? 

“There are no immediate benefits to you from your participation.” (page 40)
Attachment 1 

Page 23 of 30 

(b) What benefits to society are anticipated from the information likely to be gained 

through the research? 

“The principal beneficiary will likely be the Sponsor, for whom new data and new 

labeling will meet current U.S. EPA registration standards...For the general public, 

insect-borne disease is of growing significance in the United States and around the world 

where U.S. citizens are active. Moreover, discomfort associated with nuisance biting 

restricts many work and pleasure activities.” (p. 10) 

“[B]y serving as a participant, you may assist in making new insect repellent products 

available to consumers.” (page 40) 

(c) How would societal benefits be distributed? Who would benefit from the proposed 

research? 

“The principal beneficiary will likely be the Sponsor, for whom new data and new 

labeling will meet current U.S. EPA registration standards.” (p. 10) Indirect beneficiaries 

would include those repellent users who prefer this product to other available repellents. 

(d) What is the likelihood that each identified societal benefits would be realized? 

The testing is likely to demonstrate that the formulation is effective in repelling 

mosquitoes, and thus the sponsor is likely to realize a direct benefit from the research. 

Realization of other societal benefits will depend on consumer acceptance of the 

formulation. 

6. Risk/Benefit Balance 

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

to subjects or to society? 

The protocol systematically reduces risks to subjects without reducing the robustness of 

the scientific design. No reasonable opportunities to further reduce subject risk have 

been overlooked. The resulting residual risk to subjects is very low. The potential 

benefits to repellent users from availability of a wider variety of effective mosquito 

repellents are likely to be realized, and make the residual risks to subjects in this 

proposed research reasonable. 

7. Independent Ethics Review 

(a) What IRB reviewed the proposed research? 

Independent Investigational Review Board, Plantation FL 
Attachment 1 

Page 24 of 30 

(b) Is this IRB independent of the investigators and sponsors of the research? Yes 

(c) Is this IRB registered with OHRP? Yes 

(d) Is this IRB accredited? If so, by whom? 

Not reported. IIRB is not listed as accredited on the AAHRPP website.

(e) Does this IRB hold a Federal-Wide Assurance from OHRP? 

Not reported. IIRB is not listed as holding an FWA on the OHRP website. 

(f) Are complete records of the IRB review as required by 40 CFR 26.1125 provided? 

Complete records of the IRB review are provided in the protocol submission. 

Satisfactory documentation of IIRB, Inc., policies and procedures and of IIRB, Inc., 

membership was submitted in addition to the protocol. 

(e) What standard(s) of ethical conduct would govern the work? 

“U.S. EPA Good Laboratory Practice Regulations (40 CFR 160); 40 CFR 26 subparts K 

and L; FIFRA §12(a)(2)(P); California State EPA Department of Pesticide Regulation 

study monitoring (California Code of Regulations Title 3, Section 6710).” (p. 1) 

8. Informed Consent 

(a) Will informed consent be obtained from each prospective subject? Yes. 

(b) Will informed consent be appropriately documented, consistent with the 

requirements of 40 CFR 26.1117? Yes. 

(c) Do the informed consent materials meet the requirements of 40 CFR 26.1116, 

including adequate characterization of the risks and discomforts to subjects from 

participation in the research, the potential benefits to the subject or others, and the 

right to withdraw from the research? Yes. 

(d) What is the literacy rate in English or other languages among the intended research 

subjects? 

100%. English literacy is a requirement for participation. 

(e) What measures are proposed to overcome language differences, if any, between 

investigators and subjects? n/a
Attachment 1 

Page 25 of 30 

(f) What measures are proposed to ensure subject comprehension of risks and 

discomforts? 

Frequent opportunities to ask questions. 

(g) What specific procedure will be followed to inform prospective subjects and to seek 

and obtain their consent? 

See pp. 16-17 of the protocol and the consent documents (pp. 33-62) 

(h) What measures are proposed to ensure fully voluntary participation and to avoid 

coercion or undue influence? 

Candidates are offered repeated opportunities to decide not to participate; participants are 

offered repeated opportunities to withdraw. Exclusion factors rule out participation by 

employees or students of the Study Director. Recruitment of alternate subjects reduces 

the likelihood that subjects might be reluctant to withdraw lest the validity of the 

investigation be compromised. 

9. Respect for Subjects 

(a) How will information about prospective and enrolled subjects be managed to 

ensure their privacy? 

“Carroll-Loye Biological Research will retain records of this study indefinitely. You may 

access your own records by contacting the Study Director. Representatives from the 

sponsor (Sam Darling), the U.S. Environmental Protection Agency (EPA), the California 

Department of Pesticide Regulation and the Independent Investigational Review Board, 

Inc. (an independent committee that reviewed this study’s ethical aspects to help protect 

the rights and welfare of study participants) may have access to all non-personal 

information collected in this study. Because of the need to release information to these 

parties, absolute confidentiality cannot be guaranteed. Any information or reports 

published as a result of this study will not identify you by name, or by any other personal 

identification.” (page. 41) 

“Results of a subject’s [pregnancy] test are only observed by one female CLBR staff 

technician and never recorded to minimize stress on a female subject testing positive, and 

minimize the possibility that other staff or subjects may become aware of the results of 

that test.” (page. 8) 

Subjects are identified by name and subject number on the “Confidential Test Subject 

Information” form. On all other data collection forms, only the subject number is used. 

Recruitment of alternate subjects provides an opportunity for discrete withdrawal without 

explanation. 
Attachment 1 

Page 26 of 30 

(b) How will subjects be informed of their freedom to withdraw from the research at 

any time without penalty? 

Subjects are so informed in the recruitment interview (p. 14) and in the consent form. 

(c) How will subjects who decline to participate or who withdraw from the research be 

dealt with? 

Subjects who decide not to participate will simply go their way. Subjects identified as 

alternates, and any who withdraw from the research, will be paid for their time (page. 

41). 
Attachment 2 

Page 27 of 30 

§ 26.1111 Criteria for IRB approval of research 

Protocol No Mas 003 (7/15/10) 

Criterion Y/N Comment/Page Reference 

(a)(1)(i) Risks to subjects are minimized by using procedures which are consistent with 

sound research design and which do not unnecessarily expose subjects to risk. 

Y 

(a)(1)(ii) Risks to subjects are minimized, whenever appropriate, by using procedures 

already being performed on the subjects for diagnostic or treatment purposes. 

N/A 

(a)(2) Risks to subjects are reasonable in relation to anticipated benefits, if any, to 

subjects, and the importance of the knowledge that may reasonably be expected to 

result. In evaluating risks and benefits, the IRB should consider only those risks and 

benefits that may result from the research (as distinguished from risks and benefits 

subjects would receive even if not participating in the research). The IRB should not 

consider possible long-range effects of applying knowledge gained in the research (for 

example, the possible effects of the research on public policy) as among those 

research risks that fall within the purview of its responsibility. 

Y 

(a)(3) Selection of subjects is equitable, taking into account the purposes of the 

research and the setting in which it will be conducted, and being particularly cognizant 

of the special problems of research involving vulnerable populations, such as 

prisoners, mentally disabled persons, or economically or educationally disadvantaged 

persons. 

Y 

(a)(4) Informed consent will be sought from each prospective subject or the subject’s 

legally authorized representative, in accordance with, and to the extent required by 

§26.1116. 

Y 

(a)(5) Informed consent will be appropriately documented, in accordance with, and to 

the extent required by §26.1117. 

Y 

(a)(6) When appropriate, the research plan makes adequate provision for monitoring 

the data collected to ensure the safety of subjects. 

Y 

(a)(7) When appropriate, there are adequate provisions to protect the privacy of 

subjects and to maintain the confidentiality of data. 

Y 

(b) When some or all of the subjects are likely to be vulnerable to coercion or undue 

influence, additional safeguards have been included in the study to protect the rights 

and welfare of these subjects. 

N/A 
Attachment 3 

Page 28 of 30 

§26.1116 General requirements for informed consent 

Protocol No Mas 003 (7/15/10) 

Criterion Y/N Comment/Page Reference 

No investigator may involve a human being as a subject in research covered by this 

subpart unless the investigator has obtained the legally effective informed consent of 

the subject or the subject’s legally authorized representative 

Y All subjects will provide legally effective 

informed consent. 

An investigator shall seek such consent only under circumstances that provide the 

prospective subject or the representative sufficient opportunity to consider whether or 

not to participate and that minimize the possibility of coercion or undue influence 

Y 

The information that is given to the subject or the representative shall be in language 

understandable to the subject or the representative 

Y Information is clearly presented in plain 

English 

No informed consent, whether oral or written, may include any exculpatory language 

through which the subject or the representative is made to waive or appear to waive 

any of the subject’s legal rights, or releases or appears to release the investigator, the 

sponsor, the institution or its agents from liability for negligence 

Y The IC contains no exculpatory 

language 

(1) A statement that the study involves research, an explanation of the 

purposes of the research and the expected duration of the subject’s 

participation, a description of the procedures to be followed, and 

identification of any procedures which are experimental 

Y pp. 33, 44, 55 

(2) A description of any reasonably foreseeable risks or discomforts to the 

subject 

Y pp. 38-39, 48-50, 58 

(3) A description of any benefits to the subject or to others which may 

reasonably be expected from the research 

Y pp. 40, 51, 59 

(4) A disclosure of appropriate alternative procedures or courses of 

treatment, if any, that might be advantageous to the subject 

Y pp. 40, 51, 59 

(5) A statement describing the extent, if any, to which confidentiality of 

records identifying the subject will be maintained 

Y pp. 41, 51-52, 60 

(6) For research involving more than minimal risk, an explanation as to 

whether any compensation and an explanation as to whether any medical 

treatments are available if injury occurs and, if so, what they consist of, or 

where further information may be obtained 

Y Compensation pp. 41, 51, 59-60 

Medical Treatment pp. 40, 50, 59 

(7) An explanation of whom to contact for answers to pertinent questions 

about the research and research subjects’ rights, and whom to contact in 

the event of a research-related injury to the subject 

Y pp. 41, 51, 59 

(a) In seeking informed consent the following information 

shall be provided to each subject 

(8) A statement that participation is voluntary, refusal to participate will 

involve no penalty or loss of benefits to which the subject is otherwise 

entitled, and the subject may discontinue participation at any time without 

penalty or loss of benefits to which the subject is otherwise entitled 

Y pp. 33, 44, 55 

(1) A statement that the particular treatment or procedure may involve 

risks to the subject (or to the embryo or fetus, if the subject may become 

pregnant) which are currently unforeseeable 

Y pp. 40, 50, 58 

(2) Anticipated circumstances under which the subject’s participation may 

be terminated by the investigator without regard to the subject’s consent 

Y pp. 42, 52, 60 

(3) Any additional costs to the subject that may result from participation in 

the research 

Y pp. 41, 51, 59 

(4) The consequences of a subject’s decision to withdraw from the 

research and procedures for orderly termination of participation by the 

subject 

Y pp. 41-42, 52, 60 

(5) A statement that significant new findings developed during the course 

of the research which may relate to the subject’s willingness to continue 

participation will be provided to the subject 

N/A 

(b) When appropriate, one or more of the following elements of information shall also be provided to each subject 

(6) The approximate number of subjects involved in the study Y p. 35, 46, 56-57 

(e) If the research involves intentional exposure of subjects to a pesticide, the subjects 

of the research must be informed of the identity of the pesticide and the nature of its 

pesticidal function. 

Y pp. 33, 44, 55 
Attachment 4 

Page 29 of 30 

§26.1117 Documentation of informed consent 

Protocol NO-MAS 003 (7/15/10) 

Criterion Y/N Comment/Page Reference 

(a) Informed consent shall be documented by the use of a written consent form 

approved by the IRB and signed by the subject or the subject’s legally authorized 

representative. A copy shall be given to the person signing the form. 

Y Consent forms pp. 33-62 

(b)(1) The consent form may be a written consent document that embodies the 

elements of informed consent required by §26.1116. This form may be read to the 

subject or the subject’s legally authorized representative, but in any event, the 

investigator shall give either the subject or the representative adequate opportunity to 

read it before it is signed; or 

Y Consent form meets requirements of 

§26.1116; procedure described in 

protocol §3.4 provides adequate 

opportunity to read it before it is 

signed. 

(b)(2) The consent form may be a short form written consent document stating that the 

elements of informed consent required by §26.1116 have been presented orally to the 

subject or the subject’s legally authorized representative. When this method is used, 

there shall be a witness to the oral presentation. Also, the IRB shall approve a written 

summary of what is to be said to the subject or the representative. Only the short form 

itself is to be signed by the subject or the representative. However, the witness shall 

sign both the short form and a copy of the summary, and the person actually obtaining 

consent shall sign a copy of the summary. A copy of the summary shall be given to the 

subject or the representative, in addition to a copy of the short form. 

N/A 
Attachment 5 

Page 30 of 30 

40 CFR 26.1125 Submission of proposed human research for EPA review

Protocol No Mas 003 (7/15/10) 

Any person or institution who intends to conduct or sponsor human research covered by §26.1101(a) shall, after receiving 

approval from all appropriate IRBs, submit to EPA prior to initiating such research all information relevant to the proposed 

research specified by §26.1115(a), and the following additional information, to the extent not already included: 

Requirement Y/N Comments/Page Refs 

(1) The potential risks to human subjects Y pp. 5-10 

(2) The measures proposed to minimize risks to the human subjects; Y pp. 5-10 

(3) The nature and magnitude of all expected benefits of such 

research, and to whom they would accrue Y p. 10 

(4) Alternative means of obtaining information comparable to what 

would be collected through the proposed research; and Y p. 5 

§1125(a) 

a discussion of: 

(5) The balance of risks and benefits of the proposed research. Y p. 10 

§1125(b): All information for subjects and written informed consent 

agreements as originally provided to the IRB, and as approved by the IRB. Y pp. 198-224 (submitted) 

pp. 33-62 (approved) 

§1125(c): Information about how subjects will be recruited, including any 

advertisements proposed to be used. Y pp. 12-14. No advertisements 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. 

Y pp. 16-17 

§1125(e): All correspondence between the IRB and the investigators or 

sponsors. Y pp. 194-340 

The following Information, to the 

Extent not already included: 

§1125(f): Official notification to the sponsor or investigator. . . that research 

involving human subjects has been reviewed and approved by an IRB. Y pp. 335-336 

(1) Copies of 

• all research proposals reviewed by the IRB, 

• scientific evaluations, if any, that accompanied the proposals 

reviewed by the IRB, 

• approved sample consent documents, 

• progress reports submitted by investigators, and reports of injuries to 

subjects. 

Y 

n/a 

Y 

n/a 

pp. 1-32 

pp. 241-328 

p. 33-62 

Initial review of new proposal 

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

• attendance at the meetings; 

• actions taken by the IRB; 

• the vote on these actions including the number of members voting 

for, against, and abstaining; 

• the basis for requiring changes in or disapproving research; 

• a written summary of the discussion of controverted issues and their 

resolution. 

Y 

Y 

Y 

n/a 

n/a 

pp. 337-339 

pp. 337-339 

pp. 337-339 

pp. 337-339 

No controverted issues 

(3) Records of continuing review activities. n/a n/a for protocols 

(4) Copies of all correspondence between the IRB and the investigators. Y Provided by investigator 

pp. 194-340 

(5) 

• A list of IRB members identified by name; earned degrees; 

representative capacity; indications of experience such as board 

certifications, licenses, etc., sufficient to describe each member’s 

chief anticipated contributions to IRB deliberations; 

• any employment or other relationship between each member and 

the institution, for example, full-time employee, a member of 

governing panel or board, stockholder, paid or unpaid consultant. 

Y 

Y 

Submitted separately 

(6) Written procedures for the IRB in the same detail as described in 

§26.1108(a) and §26.1108(b). N Submitted separately 

all information relevant to the proposed research 

specified by § 26.1115(a) 

(7) Statements of significant new findings provided to subjects, as required by 

§26.1116(b)(5). n/a n/a for protocols