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UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

WASHINGTON D.C., 20460

OFFICE OF 

PREVENTION, PESTICIDES AND TOXIC 

SUBSTANCES 

September 21, 2009

MEMORANDUM

SUBJECT: Science and Ethics Review of AEATF II Aerosol Scenario Design and Protocol 

for Exposure Monitoring 

FROM: Timothy Leighton 

 Cassi Walls, Ph.D. 

 Antimicrobials Division 

 Kelly Sherman 

 Human Research Ethics Reviewer 

TO: Norm Cook, Chief 

 Risk Assessment and Science Support Branch (RASSB) 

 Antimicrobials Division 

REF: Selim, S. (2009) Transmittal Letter, 40 CFR 26.1125 Checklist, and Primary 

Documentation: Aerosol Application Scenario: Rationale for Study Design. 

56 pp. (Volume 1) 

Selim, S. (2009) A Study For Measurement of Potential Dermal and Inhalation 

Exposure During Application of a Liquid Antimicrobial Pesticide Product Using a 

Pressurized Aerosol Can for Indoor Surface Disinfecting. Unpublished protocol 

dated August 4, 2009, prepared by Golden Pacific Laboratories for the 

Antimicrobial Exposure Assessment Task Force II under Sponsor ID AEA04 and 

GPL Study No. 070270. 203 pp. (Volume 2)

Selim, S. (2009) AEATF II Aerosol Study. Secondary Documentation: IIRB 

Communications. August 4, 2009 (Volume 3) 

Selim, S. (2009) AEATF II Aerosol Study. Standard Operating Procedures for a 

Multi-Year Antimicrobial Chemical Exposure Monitoring Program. August 4, 

2009 (Volume 4) 
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We have reviewed the referenced proposal from both scientific and ethics perspectives. 

Scientific aspects of the proposed research are assessed in terms of the recommendations of the 

EPA Guidelines Series 875 and of the EPA Human Studies Review Board. Ethical aspects of the 

proposed research are assessed in terms of the standards defined by 40 CFR 26 subparts K and L 

and the recommendations of the EPA Human Studies Review Board. 

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 6. All 

elements of required documentation are provided in the submitted protocol package. 

Volume 1 of the submitted package includes the following supporting documents—all 

considered in this review: 

• Transmittal Letter (pp. 3-4)1

• 40 CFR 26.1125 Checklist (p. 5) 

• Aerosol Application Scenario: Rationale for Study Design (pp. 6-56) 

Volume 2 of the submitted package includes the following documents—all considered in 

this review: 

• IIRB-approved protocol (pp. 3-57)2

• Product Label for test material, in English (p. 59-60) and Spanish (pp. 61-63) 

• IRB-approved Informed consent form, in English (pp. 65-74) and Spanish (pp. 

75-85) 

• IRB-approved Experimental Subject’s Bill of Rights, in English (p. 87) and 

Spanish (p. 88) 

• Subject Self-Reporting Demographic Form, in English (p. 90) and Spanish (p. 91) 

• MSDS for test material, in English (p. 93) and Spanish (pp. 94-95) 

• IRB-approved Flyer soliciting research subjects, in English (p. 97) and Spanish 

(p. 98) 

• IRB-approved Newspaper advertisement soliciting research subjects, in English 

(p. 99) and Spanish (p. 100) 

• Employer contact script and subject invitation to participate script, in English (pp. 

103-104) and Spanish (pp. 105-106) 

• Executive Summaries from ADBAC and DDAC Reregistration Eligibility 

Decision Documents (pp. 111-156) 

 

1 Most page images in the submitted volumes bear more than one page number. All page references in this review 

are to page “N of 56” in Volume 1; page “N of 203” in Volume 2; page “N of 317” in Volume 3; and page “N of 

128” in Volume 4. 

2 The protocol submitted to EPA on August 4, 2009 (pages 3-57 of Volume 2) bearing a cover page dated August 4, 

2009, is identical to the version of the protocol submitted to IIRB on July 14, 2009, and approved by IIRB on July 

27, 2009. This is documented in a September 15, 2009, letter from Hasmukh Shah (Manager, AEATF II) to Kelly 

Sherman (EPA, Office of Pesticide Programs). A copy of the letter is included in the supplemental materials. 
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• IIRB approval letter of July 27, 2009 (pp. 157-159) 

• Recruitment and Consent documents submitted to IIRB, showing IIRB-requested 

changes (pp. 160-203) 

Volume 3 of the submitted package includes documentation of communications with 

IIRB, Inc. concerning the aerosol protocol, including: 

• Protocol version transmitted to IIRB (bearing cover page dated July 14, 2009; 

subsequent pages dated July 13, 2009) (pp. 20-168)3

Volume 4 of the submitted package includes all of the AEATF II’s Standard Operating 

Procedures (SOPs) supporting the aerosol study protocol. 

In future protocol submissions, it is imperative that the AEATF assign a version date to 

protocols and consent materials. The version date must be made a permanent attribute of the 

file. The AEATF’s re-dating of the IIRB-submitted and approved protocol jeopardizes the 

integrity of the record. Please refer to the September 15, 2009, letter from Hasmukh Shah 

(Manager, AEATF II) to Kelly Sherman (EPA, Office of Pesticide Programs) for details. 

A. Summary Assessment of the Scenario Design 

Supporting details are in Attachment 1. 

1. Scenario Design: The Antimicrobials Division (AD) assesses potential occupational 

and consumer exposure from various antimicrobial products that are applied by a 

multitude of application techniques including from an aerosol spray can. Aerosol 

spraying can be accomplished with different types of nozzles, volumes, and 

techniques. AEATF II defines “...the aerosol application scenario is defined as the 

hand-held pressurized aerosol-based application of a label-specified end-use 

formulation containing an antimicrobial chemical.” (V1:8)4 The actual task of 

spraying will be to spray a smooth, sweeping, overlapping pattern until the treated 

surface is wet. The scope of the choices of aerosol can characteristics that are 

reasonably expected to influence exposure includes nozzle size, particle size 

generated, ejection rate, and generation of non-volatile active ingredient. From this 

array of choices, the industry and regulatory consensus was to monitor the selected 

product – Clorox Commercial Solutions Clorox Disinfecting Spray (EPA Reg. No. 

67619-03). A more detailed description of the process used to select the product is 

 

3 The protocol submitted to EPA on August 4, 2009 (pages 3-57 of Volume 2) bearing a cover page dated August 4, 

2009, is identical to the version of the protocol submitted to IIRB on July 14, 2009, and approved by IIRB on July 

27, 2009. This is documented in a September 15, 2009, letter from Hasmukh Shah (Manager, AEATF II) to Kelly 

Sherman (EPA, Office of Pesticide Programs). A copy of the letter is included in the supplemental materials. 

4 This pagination convention is used throughout this review. “V1” refers Volume 1 of the AEATF submission; 

“V2” refers to Volume 2; “V3” refers to Volume 3; “V4” refers to Volume 4. Entries after the colon are page 

references; many page images bear more than one page number. In Volume 1, the cited page number is always from 

the expression “p. N of 56”. Volume 2 page references are always from the expression “p. N of 203.” Volume 3 

page references are always from the expression “p. N of 317”. Volume 4 page references are always from the 

expression “p. N of 128.” 
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provided below in Attachment 1 and Appendix A Study Rationale Product Selection 

Justification. (V1:34) 

EPA intends to use the data developed by the AEATF II for the aerosol spraying 

scenario to describe a typical occupational handler’s daily exposure to the 

antimicrobial spray. The data must be generic enough to be useful for estimating 

exposures using various types of aerosol spray products (i.e., disinfecting sprays onto 

surfaces, air sanitizers, and foaming aerosols for bath tub/toilets), different types of 

surfaces, room configurations, types of buildings (i.e., homes, schools, medical 

facilities, commercial office buildings, etc.), handlers (i.e., professional and 

consumer), and antimicrobial active ingredients. AD plans to use the data generated 

from the proposed aerosol study generically to estimate dermal and inhalation 

exposures and risks for other antimicrobial ingredients where the product is packaged 

in a pressurized aerosol spray can. 

Antimicrobial products have been grouped by EPA into the 12 Use Categories listed 

below as presented in the proposed 158W data requirements. Although aerosol 

spraying of surfaces or air sanitizers may occur in many Use Categories, most aerosol 

spray applications are expected to occur in Use Categories II, III, IV, and V. 

I Agricultural premises and equipment 

II Food handling/storage establishments/premises and equipment 

III Commercial/institutional/industrial premises and equipment 

IV Residential and public access premises 

V Medical premises and equipment 

VI Human drinking water systems 

VII Material preservatives 

VIII Industrial processes 

IX Antifouling coatings 

X Wood preservatives 

XI Swimming pools 

XII Aquatic areas 

EPA believes that the AEATF II aerosol scenario is well defined, and we expect that 

resulting data will meet the needs of EPA and other regulatory agencies. The 

diversity of daily exposures under the aerosol scenario as defined in this proposal will 

adequately describe a typical occupational handler’s daily exposure to the 

antimicrobial application. This exposure can then be extrapolated to the likely 

exposure of a home user of aerosol antimicrobial products. 

2. Sampling Design: The AEATF II has described in detail their sampling design for 

the aerosol scenario and has incorporated random elements where feasible. The 

AEATF II proposes to monitor dermal and inhalation exposures using passive 

dosimetry techniques to measure exposure of human subjects spraying indoor 

surfaces with an aerosol can. The proposed sample size is 3 clusters (sites) with 6 

subjects at each. This is believed adequate to provide data to meet EPA’s needs. 
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The aerosol scenario will consist of professional applicators using 19-ounce 

pressurized cans to spray aerosols on surfaces such as laminate, tile, porcelain, glass, 

and/or metal. The physical aspects of spraying include holding the spray can, 

depressing the nozzle with a finger, and spraying in a smooth, sweeping, overlapping 

pattern. The surfaces will be sprayed from a distance of 6 to 10 inches until the 

surface becomes visibly wet. The aerosol spray cans are ready-to-use (i.e., no 

mixing/loading is involved.). Additionally, the spray solution will be left on the 

surface, so no wiping will occur. 

The amount of product applied during each monitoring event will be varied from 1 to 

4 cans. MEs will be based on 1⁄2 -can increments (i.e., 1 to 1.5 cans, 1.5 to 2 cans, up 

to 3.5 to 4 cans). The spraying duration will consist of the time it takes to spray a 

given number of cans. The focus on professional applicators is a practical necessity, 

given that residential consumer spraying efforts are of shorter duration (i.e., less 

product sprayed) and would have a greater chance that samples would be at or below 

the limit of detection of the analytical methods used. Because of longer task duration 

and consequent exposure to greater quantities of antimicrobial, professional spray 

applicator exposure is expected to be greater than that of consumers. 

The AEATF II aerosol spray study is further limited to application using a surface 

disinfecting spray. Other registered aerosol spray uses include air sanitizers and 

foaming aerosols (e.g., bath tub and toilet cleaners). The surface disinfecting spray 

was selected to represent the high end of potential exposure, based on a pilot study 

reported in the protocol submission (V1:34). The AEATF II, in consultation with 

EPA/CDPR/PMRA, considered the various configurations of aerosol spraying and the 

results of the pilot studies’ nozzle size, particle size generated, and amount of 

material dispensed per unit time, air concentration, and ejection rate, and agreed that 

the hard-surface disinfection spray application representing the reasonable high end 

of exposure. Other key factors considered in the selection of an aerosol application 

included its common use among consumers; its use in tight spaces such as shower 

stalls; and the high exposure potential for applicators walking into sprayed mist while 

spraying a surface disinfectant in small rooms. 

The scenario does not include wiping the sprayed product from surfaces that have 

been sprayed, for two reasons. First, some antimicrobial sprayed products are leaveon, not requiring the user to wipe the surface. Secondly, the AEATF II is developing 

other data under the “wipe” scenario which can be considered along with the results 

of this study when the situation warrants. This will allow the regulatory agencies to 

assess either leave-on products or those products that are subsequently wiped from 

surfaces. 

The amount of product to be handled by each subject will be randomly selected. The 

subjects will be randomly ordered as they are enrolled. At each site the subject’s 

tasks are defined by a specific amount of product to be applied ranging from 1 to 4 

cans. Set durations will not be established. Subjects will apply the amount of 
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product assigned to them as they normally would work. The first subject randomly 

selected will be assigned to the task with the largest amount to be applied (i.e., 3.5 to 

4 cans), although if the subject is unable to complete the assigned amount to be 

sprayed, the monitoring event may fit the specification for a lesser amount sprayed. 

As additional subjects are assigned to tasks, all defined spraying amounts will be 

reflected in one monitoring event. The actual duration of the spraying task will vary 

among the subjects; the AEATF II anticipates the monitoring events will range from 

30 to 180 minutes. 

The study location has been purposively selected to be Fresno County, CA. EPA 

believes that the application of an antimicrobial product in an indoor environment 

will not vary substantially from one city to another, and therefore that the selection of 

Fresno County is reasonable. Fresno is close to the laboratory conducting the study, 

it is large enough to have a substantial population of professional janitors, and it 

contains many suitable vacant buildings. Conducting all monitoring in a single 

geographic area will save resources and will not adversely effect the results of the 

research. 

The aerosol scenario design calls for three clusters of MEs, each at a different site. A 

site is defined as the facility (i.e., building). The protocol specifies further that each 

site must be a different type of building, that monitoring at different sites must be 

separated by at least one week, and that each selected building must include 

appropriate building/room configurations, specifically hotels/motels with 20 or more 

units containing various full kitchens, kitchenettes, or bathroom-only configurations. 

The need for multiple rooms within a building differs from the previous AEATF II 

mop and wipe protocols. The multiple rooms are required in this study because it is 

believed that the smaller rooms with multiple surfaces to be sprayed such as shower 

fixtures, toilets, sinks found in typical bathrooms in hotels/motels will lead to the high 

end of potential exposures occurring from aerosol applications of antimicrobial 

chemicals. 

Three sites will be selected using a stratified random sampling approach. A listing of 

potential sites will be developed using YellowPages.com. This list will be based on a 

search using the following criteria: “hotel or motel with kitchenette or full kitchen” 

in “Fresno, California”. A caveat is added to the protocol stipulating that if motels 

with kitchens are not available the search will be broadened to include empty 

apartments. The three sites will be randomly selected from this list. The three sites 

will need to meet the following selection criteria: 

• Willing to cooperate in the study; 

• Configured for a diversity of treatment surfaces (e.g., kitchens, bathrooms, 

sinks, countertops, toilets); 

• Has a functional HVAC system; 

• Has operating electrical service; 

• Does not require specialized cleaning or maintenance before use. 
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The final selection of the three sites will be those facilities that fit into one of the 

following three categories (i.e., one site per each of the three categories): 

• Hotel/motel with full kitchen and 20 or more units; 

• Hotel/motel with kitchenette and 20 or more units; 

• Hotel/motel without kitchen/kitchenette and 20 or more units. 

The aerosol scenario calls for monitoring 6 subjects at each of three sites as they 

spray horizontal and vertical surfaces. Therefore, a total of 18 test subjects will be 

monitored. Two additional test subjects will be enrolled for each site, making a total 

of 24 subjects in all, in case some subjects withdraw or fail to meet all eligibility 

criteria. Subjects will be professional janitors, selected from among those who 

respond to flyers posted in randomly selected janitorial service companies in the 

Fresno area, and who meet the eligibility criteria for participation. In addition to the 

flyers, advertisements will be run simultaneously in three newspapers: (1) Fresno 

Bee, (2) California Advocate, and (3) Vida en el Valle. Enrolled subjects will be 

randomly assigned to spraying tasks of differing amounts of product at one of the 

three test sites. 

In order to make the most effective use of a limited number of monitoring events, the 

overall sampling design is purposive. No feasible opportunities to incorporate 

random elements in the sampling design, however, have been overlooked. 

3. Choice of Surrogate Material: The choice of ADBAC as the antimicrobial material 

for this scenario is appropriate. It is widely used, readily available, and there is a 

reliable and sensitive analytical method available for it. 

C. Summary Assessment of the Scientific Aspects of the Study Design 

Supporting details are in Attachment 2.

1. Statistical design: “AEATF II proposes for the aerosol scenario a design of 3 clusters 

of 6 monitoring events (MEs) each, as discussed in the Governing Document. The 

AEATF II reviewed two data sets in determining the statistical design. The two data 

sets include aerosol-specific exposure data from PHED and the data set presented in 

the Governing Document for repetitive-motion tasks. The existing data set for 

aerosol exposure data in PHED indicates less variation then the larger data set of 

repetitive-motion tasks. The existing PHED aerosol-specific data indicate the need 

for a 3 clusters of 2 MEs to achieve the 3-fold accuracy goal. However, ...“[g]iven 

the sensitivity of the sample size to GSD [geometric standard deviation] and the 

belief that the repetitive-motion GSD is a better indicator of the expected true relative 

variation for this scenario, the AEATF II prefers to assume GSD=2.9 for the purposes 

of determining sample size.” (V1:28, 29) This assumption is consistent with the 

proposed design of 3 clusters of 6 MEs each. 
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2. Proposed pattern of human exposure: The AEATF II proposes to select 

professional janitors as subjects for the aerosol scenario. The test subjects will use 

Clorox Commercial Solutions Clorox Disinfecting Spray (EPA Reg. No. 67619-03) 

in 19 ounce ready-to-use pressurized spray cans, and each subject will spray from 1 to 

4 cans. The duration of spraying will be based on the amount of product sprayed, the 

room configuration, and the normal work habits of the test subject. The protocol 

anticipates the duration to range from 30 to 180 minutes. The actual spraying 

mechanics are to hold the spray can 6 to 10 inches from the surface, then to hold the 

nozzle down with a finger and spray overlapping swaths until surfaces are wet. The 

test subjects will not wipe the solution from the surface – the wiping task will be 

monitored in a separate AEATF II study. 

Clorox Commercial Solutions Clorox Disinfecting Spray which contains the active 

ingredients didecyl dimethyl ammonium chloride (DDAC) at a concentration of 

0.0945%, n-alkyl (40% C12, 50% C14, and 10% C16) dimethyl benzyl ammonium 

chloride (ADBAC) at a concentration of 0.252%, octyl decyl dimethyl ammonium 

chloride (ODAC) at a concentration of 0.0945%, and dioctyl dimethyl ammonium 

chloride (DODAC) at a concentration of 0.189%, and ethanol at a concentration of 

65% (plus 34.37% inerts). The C14 portion of ADBAC is the active ingredient to be 

measured on the dosimeters. The aerosol product is ready-to-use so it will be sprayed 

at the concentrations listed above with no dilution. The concentration of the test 

material will be constant for all subjects. Variation in exposures will result from 

differing amounts handled by each subject. 

The EPA believes that the AEATF II aerosol study will represent typical worker 

methods of applying the test substance to indoor surfaces. The selection of janitorial 

subjects, test material, and spraying activities as described in the protocol is justified. 

3. Endpoints and Measures: The AEATF II proposes to measure dermal and 

inhalation exposures resulting from spraying of indoor surfaces. Dermal and 

inhalation exposure will be measured using whole-body dosimeters (inner and outer), 

hand and face washes, and personal air monitors. The Agencies are most interested in 

the inner dosimeters to assess potential exposure. The outer dosimeters will add to 

the existing data base on the development of protection factors for single layer of 

clothing. The potential for foot exposure is minimal and the feet will not be 

monitored. The hand and face wash is an appropriate method to determine exposure 

to the hands and face. The personal air samplers will collect residues from the 

breathing zone with the sampling cartridge facing downwards (mimicking nostrils). 

The sampling train will consist of OVS tubes with glass filters backed by XAD2 

sorbent. Because the air sampling will not size particles, the Agencies will assume 

conservatively that all of the residues are inhalable and/or respirable, which will tend 

to overestimate inhalation exposure. The sampling pumps will be calibrated prior to 

use. 
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Environmental parameters of the work area such as air temperature, and relative 

humidity will also be documented. The HVAC system will be described and the air 

turnover rates will be measured or estimated. 

4. QA/QC Plan: The AEATF II QA/QC plan for the aerosol study is described in 

sufficient detail and is adequate to ensure that the measurements are accurate and 

reliable. The QA/QC plan includes field recovery analyses, travel recovery analyses, 

storage stability studies, and break-through analyses of the air samplers. Method 

validation results should be included in the final study report. 

Primary components of the field recovery analyses include: two fortification levels 

per matrix at the LOQ, triplicate samples per fortification level, exposed to ambient 

conditions for the maximum duration of exposure, and WBD not covered during 

exposure duration. Field recovery samples will be fortified in the field and 

transported and stored in the same was as the actual study samples, and will be 

analyzed concurrently with the actual exposure samples. Correction for loss in field 

recoveries will correct for all phases of potential losses. 

A storage stability study is ongoing. The storage stability samples are being stored 

for up to 6 months. 

5. Statistical Analysis Plan: The results of physical sample analysis will be provided 

in the final report. The AEATF II will not statistically analyze the monitoring data. 

However, the aerosol spray monitoring data will be imported into the AEATF II 

database (BHED®) where they will be available to regulatory agencies for later 

statistical analyses. The study documentation will report a confidence-interval-based 

approach to determine the relative accuracy for the arithmetic mean and 95th 

percentile of exposure normalized by amount of ai handled. The report will further 

provide the intraclass correlation for clusters (ICC) and its confidence interval, using 

a variance components model. In addition, the effects of ignoring clusters in the 

estimation of means and percentiles will be determined by comparing the estimates of 

a no-cluster model to those of the random effects model. 

D. Compliance with Applicable Scientific standards 

This protocol adequately addresses the following elements according to applicable 

scientific standards: 

• Scientific objective 

• Experimental design for achieving objectives 

• Quantification of the test materials 

• Data collection, compilation and summary of test results 

• Justification for selection of test substance and dilution rate 

• Justification for sample size 

• Fortification levels and number of samples for laboratory, field, and storage stability 

samples 
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Additionally, the AEATF II has addressed the technical aspects provided in the 

applicable exposure monitoring guidelines (i.e. Series 875 Group A and OECD Applicator 

Guidelines) as well as Good Laboratory Practices (GLPs). 

The following element in the protocol requires revision before the research goes forward: 

• The AEATF II needs to revise the protocol to indicate the course of action if the 

benchmark accuracy goals (i.e., k=3) are not achieved. 

- In a September 18, 2009, letter from Hasmukh Shah (Manager, AEATF II) to 

Timothy Leighton (Office of Pesticide Programs, Antimicrobial Division), the 

AEATF II committed to address deviations from the benchmark accuracy goal 

(i.e., k=3) as follows: “[I]f large deviations from the benchmark goals are 

observed in the aerosol study, AEATF II will, in consultation with regulatory 

agencies, determine the best course of action to take. This may mean the 

development of guidance for the use of these data that takes the increased 

imprecision of the estimates into account. It is possible that collection of 

additional clusters might be considered.” (a copy of the letter is included in the 

supplemental materials provided for review by the HSRB). 

E. Summary Assessment of Ethical Aspects of the Proposed Research 

Supporting details are in Attachment 2. 

1. Societal Value of Proposed Research: The purpose of the proposed monitoring 

study is to develop more accurate information on worker exposures to liquid 

antimicrobial products applied using pressurized aerosol cans for indoor surface 

disinfecting. Because many millions of Americans apply antimicrobial products 

using aerosol cans, the research question is important; it cannot be answered with 

confidence without new monitoring data meeting contemporary standards of quality 

and reliability. 

2. Subject Selection: Twenty-four adult subjects will be recruited among professional 

janitorial workers of Fresno County. Participants will self-identify in response to 

flyers posted in the workplaces of janitorial services companies and advertisements in 

three Fresno newspapers targeting different demographic groups. 

 The use of newspaper advertisements is a new recruiting method adopted by the 

AEATF II because workplace flyers were ineffective during recruiting for the mop 

and wipe studies. Although more than 50 flyers soliciting subjects for the mop and 

wipe studies were distributed to janitorial firms in Fresno County, no one attended 

either janitorial firm manager meeting held to explain the study. Many janitorial 

firms that reviewed the flyer declined to post, and there were very few responses from 

employees of those employers who did agree to post a flyer. In June 2009, the 
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AEATF II amended the mop and wipe protocols to allow for the use of newspaper 

advertisements in addition to workplace flyers for recruiting. Since the amendment, 

subjects have been recruited for the mop and wipe studies through both methods. The 

AEATF II intends to employ both recruiting methods simultaneously for the aerosol 

study. 

Callers responding to either the workplace flyers or the newspaper advertisements 

will be screened, scheduled for informed consent meetings, and enrolled at the 

volunteer’s convenience. Although additional randomization could be obtained if 

candidates for informed consent meetings and enrollment were randomly selected 

from a pool of respondents, the AEATF II has concluded that it is necessary to enroll 

subjects as they are processed. The AEATF II learned in recruiting for the mop and 

wipe studies that delaying enrollment in order to compile and randomize a list of 

potential subjects is infeasible because it results in significant attrition among 

potential participants and delays in the recruiting process. 

Participants will be screened from among respondents to either the workplace flyers 

or the newspaper advertisements, and in that sense will be representative of the 

population of janitorial service workers in the Fresno area. There is no reason to 

think that janitorial service workers in Fresno are not typical of janitorial service 

workers in any other area of the United States. 

Inclusion/exclusion criteria are complete and appropriate. Pregnant or nursing 

women are excluded from participation. Employees or relatives of employees of the 

investigators and of cleaning product manufacturers are also excluded from 

participation. 

No potential subjects are from a vulnerable population. Recruitment materials and 

interactions with potential subjects will be conducted in English or Spanish, 

depending on subject preference. To minimize the potential for coercion or undue 

influence from their employers, subjects will not be recruited directly through 

contract janitorial service companies. Flyers posted in workplaces will instruct 

interested workers to contact the investigators directly. 

3. Risks to Subjects: The proposed test material is EPA-registered for the use 

proposed, is of low toxicity to mammals, and will be used in full compliance with the 

approved label. Risks to subjects include risks of a reaction to the test material or the 

solvents used to obtain residues from hands and face/neck; of discomfort and possibly 

heat-related illness associated with wearing two layers of clothing while doing 

physically demanding work; of discomfort or inconvenience from wearing the air 

sampling device; of embarrassment from disrobing in the presence of a research 

technician; of an unexpected result of pregnancy testing. All identified risks are 

characterized as of low probability. 

Risks are minimized by exclusion of candidates known to be sensitive to quaternary 

ammonium compounds or in poor health or with broken skin on hands, face, or neck; 
Page 12 of 52 

testing in a controlled-temperature environment; alerting subjects to signs and 

symptoms of heat stress; monitoring heat index with associated stopping rules; 

limited time of exposure with rest periods at 30-minute intervals, or more frequently 

if requested; close observation of subjects; training of experienced technicians to 

minimize embarrassment; incorporation of procedures to keep results of pregnancy 

testing private and to permit discrete withdrawal; provision of appropriate work 

clothing and PPE. 

4. Benefits: This research offers no direct benefits to the subjects, but subjects may 

request their individual results, from which they may learn that their work practices 

produce more or less exposure than average. The principal benefit of this research is 

likely to be reliable data about the dermal and inhalation exposure of workers 

applying antimicrobials in pressurized aerosol cans, usable by EPA and other 

regulatory agencies to support exposure assessments for a wide variety of 

antimicrobial products and their uses. 

5. Risk/Benefit Balance: Risks to subjects have been thoughtfully and thoroughly 

minimized in the design of the research. The low residual risk is reasonable, in light 

of the likely benefits to society from new data supporting more accurate applicator 

exposure assessments for antimicrobial products applied with pressurized aerosol 

cans. 

6. Independent Ethics Review: The proposed research has been reviewed and 

approved by the Independent Investigational Review Board, Inc., (IIRB) of Plantation 

Florida. The submitted materials include a full record of correspondence between the 

investigators and the IIRB.

7. Informed Consent: Informed consent will be obtained from each prospective 

subject and appropriately documented in the language preferred by the subject. 

Literacy in English or Spanish is a requirement for inclusion. 

All written recruitment, consent, and risk communication materials will be available 

in both English and Spanish. In order to ensure effective communication and 

thorough comprehension by anyone preferring Spanish over English, a Spanishspeaking member of the research team will be present at the meetings at which 

candidates are qualified and sign consent forms. 

8. Respect for Subjects: Subject-identifying information will be recorded only once; 

all subsequent data records and reports will refer to individual subjects only by an 

arbitrary code. Provision is made for discrete handling of pregnancy testing, required 

of female subjects on the day of testing. Candidates and subjects will be repeatedly 

reminded that they are free to decline to participate or to withdraw at any time for any 

reason, without penalty. 
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F. 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, and because the research will be conducted in California, the 

provisions of California Code of Regulations Title 3, Section 6710 apply as well. 

A detailed evaluation of how this proposal addresses applicable standards of ethical 

conduct is included in Attachments 2-5 to this review. 

The following specific deficiency in the protocol should be addressed before the research 

is initiated: 

• In the statement about compensation for research-related injuries (p. 7 of 10 of the 

consent form), please make the following clarifying revision: 

o Current language: “We will pay for needed medical treatment that is not paid 

for by your own insurance or by someone else.” 

o Change to: “We will pay for needed medical treatment that is not paid for by 

your own insurance or by the insurance of a third party under which you are 

covered.” 

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. 

The 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. 

 If the deficiency noted above is addressed and the amended protocol is approved by the 

overseeing IRB, this research should meet the ethical standards of FIFRA §12(a)(2)(P) and 40 

CFR 26 subparts K and L. 
Page 14 of 52 

Attachments: 

1. Summary Review of Aerosol Scenario Design dated August 4, 2009 

2. Summary Review of AEATF-II Protocol AEA04 dated August 4, 2009

3. §26.1111 Criteria for IRB approval of research 

4. §26.1116 General requirements for informed consent 

5. §26.1117 Documentation of informed consent 

6. §26.1125 Criteria for Completeness of Proposals for Human Research 
Attachment 1 

Page 15 of 52 

EPA Scenario Review: AEATF-II Aerosol Application Scenario/Protocol 

Title: AEROSOL APPLICATION SCENARIO: RATIONALE FOR STUDY 

DESIGN (AEATF-II Volumes I and II) 

Date: August 4, 2009 

Sponsor: American Chemistry Council 

Antimicrobial Exposure Assessment Task Force II 

c/o Hasmukh Shah, Ph.D. 

1300 Wilson Blvd 

Arlington VA 22209 

 

1. Scope of Scenario Design 

(a) Is the scenario adequately defined? 

Preliminary versions of the aerosol field study protocol have been reviewed by EPA, 

PMRA, and CDPR to determine the appropriate aerosol scenario to assess exposure as 

accurately as possible, while ensuring that any error does not underestimate exposure. 

Many varied antimicrobial products can be applied by hand-held pressurized aerosol cans 

for air and surface odor elimination, and for sanitizing and disinfecting hard or soft 

surfaces. Aerosol applications are common in residential (consumer) settings as well as 

in commercial settings such as hotels, hospitals, schools, or restaurants. 

Aerosol choices include hard and soft surface disinfecting sprays, air sanitizers, and 

foaming aerosols. From this array of choices, the consensus among the Task Force and 

the three regulatory agencies was to select the pressurized disinfecting aerosol spray for 

hard surfaces. The hard surface “pressurized disinfecting aerosol spray was selected to 

represent this group of spray products because it is a commonly used product type, the 

application process (spray nozzle actuation) is similar across all aerosol product types, 

and the surface application rates, normalized to mass of formulation per surface area are 

comparable across product types.” (V1:8) 

The AEATF II aerosol protocol appropriately proposes to use Clorox Commercial 

Solutions® Clorox® Disinfecting Spray (EPA Reg. No. 67619-03), based on the nozzle 

size, amount of material dispensed per unit time, air concentrations, and aerosol 

characteristics, across products in the four major aerosol categories (hard surface 

disinfecting sprays, hard surface foaming aerosols, soft surface sprays, air fresheners). 

“The aerosol application scenario involves application according to typical practices, e.g., 

spraying surfaces from a distance of approximately 6-10 inches in a manner to apply 

enough formulation to provide an adequate amount for cleaning. Hard surface 
Attachment 1 

Page 16 of 52 

applications are typically sprayed until visibly “thoroughly” wet per label direction.”

“Surface applications are typically made in smooth, sweeping, overlapping patterns. 

Examples of ‘representative’ spray application techniques that this scenario is expected to 

capture include horizontal spraying moving upward and downward from the starting 

point to hard surfaces such as laminate, tile, porcelain, glass, and metal).” (V1:8) 

Aerosol applications may or may not involve wiping the sprayed surface. Some aerosol 

products are labeled as “leave-on” sprays that do not require post-spray wiping. Only the 

actual aerosol spray application is covered by this scenario; the exposure from postapplication wiping, such as a potable water rinse and wipe following aerosol application 

to a food preparation surface, will not be evaluated in this scenario. Wiping exposure will 

be monitored in a separate AEATF II study. This separation of tasks will allow 

regulatory agencies to assess the exposure associated with either professional “leave-on” 

products or other products requiring post-spray wiping. 

The AEATF II study proposes to recruit only professional janitors. Janitors have been 

selected to provide longer task durations then are typical for consumers, to reduce the 

likelihood of undetectable residues on sampling materials. Thus, the scenario design will 

generate conservative data when used to estimate consumer application exposures. 

The amount of product applied during each monitoring event will be varied from 1 to 4 

cans. MEs will be based on 1⁄2 -can increments (i.e., 1 to 1.5 cans, 1.5 to 2 cans, up to 3.5 

to 4 cans). The spraying duration is not predetermined. The duration will consist of the 

time it takes to spray a given number of cans and is anticipated to range from 30 to 180 

minutes. 

(b) Is there a need for the data? Will it fill an important gap in understanding? 

PHED contains two studies conducted using aerosol application methods. These studies 

have limitations that reduce their value for an antimicrobial-oriented generic database. 

Both of these studies involved aerosol application for indoor residential crack and crevice 

insecticide treatment which may not be representative of antimicrobial use patterns. 

Although both studies monitored application in different houses, the same subjects were 

used for multiple MEs. Every ME within a study applied an identical amount of product. 

Thus, there is no variation in amount of a.i. handled within a study and very little 

difference between the two studies (V1:11). Lastly, these studies do not report particle 

size distribution. 

Furthermore, all subjects in study 456 wore chemical resistant gloves, a practice that is 

not typical for aerosol application of antimicrobials. In addition, both the dermal and 

inhalation exposure data from this study have low quality analytical data; as a result, 

study 456 has rarely been considered in regulatory exposure assessments (V1:11). 

Data from the CMA study for aerosol application include only 5 monitoring events with 

measurable exposures, and only hand exposures were detectable for these MEs. The 

application duration, only a fraction of which involved actual aerosol spraying, ranged 
Attachment 1 

Page 17 of 52 

from 9 to 260 minutes. Finally, the regulatory agencies agreed that the limited number of 

replicates combined with poor recovery data severely limits the conclusions that can be 

made from the CMA study (V1:12). 

Based on the PHED and CMA data limitations, the Antimicrobial Division is requiring 

dermal and inhalation exposure data in many of its assessments to fill this data gap for 

aerosol products. 

2. Rationale for Scenario Sampling Design 

(a) Are the variables in the aerosol scenario design likely to capture diverse exposures 

at the high-end? 

The design choices in the pressurized aerosol scenario to provide diversity in sampling 

include (1) application equipment; (2) multiple indoor sites in one geographic location; 

(3) varying the amount of active ingredient handled by the subjects; and (4) using 

different workers for each monitoring event. 

Application equipment. The AEATF II evaluated several different types of products 

and equipment (i.e., spray/can) parameters in order to select the most representative 

and appropriate aerosol spray product for use in the study. This discussion 

summarizes the AEATF II’s aerosol product/can selection process selection process; 

Appendix A of Volume 1 presents the process in complete detail (V1:34). 

 

The selected aerosol product needs to meet the following criteria (V1:38): 

• Serve as surrogate for most aerosol use categories where results can be 

extrapolated to most products on the market 

• Use pattern represents high end exposure 

• Use scenario covers most influential variables of exposure 

• Have stable active ingredient 

Based on a review of the existing literature and exposure data for pressurized aerosol 

products, the following variables were identified as having the most impact on 

potential exposure (V1:38): 

• Amount of material used 

• Release rate 

• Particle size distribution 

• Nozzle technology 

• Pressure in the can 

• Temperature / humidity 

• Surface on which product is used 

• Orientation of the can during use 

 AEATF II aerosol product/can selection process was conducted in three phases. The 

first phase consisted of conducting an internal membership survey that identified all 

of the members’ aerosol products and characterized them by some of the influential 
Attachment 1 

Page 18 of 52 

exposure variables. The next phase categorized each of the products into one of four 

major use scenarios. The final phase consisted of a method development study where 

one representative product from each of the four categories was selected for further 

analysis on the influencing variables. Based on the results of the method 

development study, one of the four products was selected as the representative 

aerosol product for use in the overall exposure study. 

Results of the phase 1 AEATF II survey identified 18 products marketed by 9 major 

companies used in both household and commercial/institutional settings. Many 

products contain more than one active ingredient; 10 of the 18 products, however, 

contain ADBAC. Nozzle size, release rates, and particle size parameters were 

compiled for each of the 18 products. If the release rates or particle sizes were not 

available from the company, the data were experimentally obtained. “The data show 

that the release rates range from 0.66 g/second – 2.8 g/second and the nozzle sizes 

range from 0.013 – 0.03 inches, and most of the nozzles sizes were ~0.02′′. The 

release rate and particle size is dependent on the combination of nozzle technology 

and can characteristics. Aerosol can sizes range from 2.5 -19 oz and are pressurized 

at 40-60 PSI with the propellant of choice. . . . The particle size distribution data 

ranged from 16 -164 μm depending on the product type and method of particle size 

determination. The hard surface fine spray products had particle size distribution 40-

157 μm. The surface disinfectants which are also used as air fresheners/sanitizers and 

air treatment products had particle size range of 16-87 μm, and most of them had the 

smallest nozzle size. The foaming spray products had particle size distribution of 24-

164 μm” (V1:43). 

The targeted surface type (i.e., hard vs. soft surface) and the orientation of the can are 

variables that were also considered in the product selection process. “A hard target 

surface with a vertical and overhead orientation of the spray can was considered the 

most conservative scenario (highest exposure use) and is expected to give the most 

bounce- back from the spray that increases the air concentration in the breathing zone 

and dermal deposition.” (V1:43) 

The second phase of the selection process placed all of the surveyed antimicrobial 

aerosol products into the following four major use categories: 

• Hard – surface disinfectant fine sprays 

• Foaming aerosol products – coarse sprays 

• Soft surface disinfectants 

• Air fresheners 

In the final phase of the product selection process, one representative product from 

each of the four categories was selected for inclusion in the pilot study (i.e., further 

analysis on its influencing parameters (i.e., release rate and particle size distribution)) 

for selection as the overall representative aerosol product in the exposure study. 

“The results of the pilot study clearly show that based on nozzle size, amount of 

material dispensed per unit time, air concentrations, and aerosol characteristics, the 
Attachment 1 

Page 19 of 52 

hard surface disinfectant product, i.e., Clorox Disinfecting Spray (EPA Reg. No. 

67619-03), represents the high-end exposure scenario and the product most likely to 

produce measurable exposure and would therefore serve as the surrogate for the study 

entitled, “Measurement of Potential Dermal and Inhalation Exposure During 

Application of a Liquid Antimicrobial Pesticide Product Using a Pressurized Aerosol 

Can for Indoor Uses”. Taking type of surface and the inspirable mass into 

consideration, the Clorox Disinfecting Spray (hard surface fine Spray) and Stepan 

Aerosol SDAS (Air Freshener) represent uses with the most inhalation and/or dermal 

exposure potential. The Air Freshener seemed to have comparable inspirable mass to 

the hard surface spray. However, as mentioned previously, the total mass of the active 

ingredient dispensed per unit time for the Clorox Disinfecting Spray is more than 

double the Stepan Aerosol SDAS (17.2/6.7 mg, Table 4) and dermal exposure will 

likely be higher for the Clorox Disinfecting Spray. Additionally, the hard surface 

spray product will be used to a much greater extent in a day, especially in commercial 

use, than the air freshener. Based on the available data, the Clorox Disinfecting Spray 

(hard surface fine spray) would represent a high end conservative choice for exposure 

monitoring studies” (V1: 55). 

Sites. Three sites (i.e., vacant commercial lodging facilities or vacant areas and 

rooms within otherwise occupied buildings) in Fresno County, CA, will be selected 

for this study. “Commercial lodging facilities are buildings that are most likely to 

provide an adequate amount of relevant aerosol application surface area for the 

monitoring events. While other building types, such as offices and meeting locations 

represent locations where disinfecting aerosols may be applied, and provide diversity in 

architecture and floor plan, these categories are less likely to provide the number of 

separate rooms and surface areas needed for the range of amount of aerosol to be 

sprayed” (V1:19). 

“...Greater diversity among monitoring sites can be obtained if each of the three 

clusters is conducted in a somewhat different room configuration. Consequently, for 

the aerosol application study the AEATF II will consider only the following three 

building/room configuration categories: 

A. Hotels/motels with 20 or more available units containing full kitchens 

B. Hotels/motels with 20 or more available units containing kitchenettes 

C. Hotels/motels with 20 or more bathroom-only units. 

These three categories were chosen because they vary with respect to bathroom and 

food preparation areas (i.e., kitchen or ‘kitchenette’) configurations which might be 

expected to impact exposure potential differently” (V1:30). 

The choice of one geographical location of Fresno County assumes indoor aerosol 

application tasks will not differ substantially from one geographic location to another. 

This assumption is supported by the previous CMA study. Moreover, the available 

training material for janitorial practices is supported by national organizations (e.g., 

International Sanitary Supply Association (ISSA)), and regional differences are not 

expected. The choice of sampling vacant buildings allows the researchers “...to be 

free from personal interferences with non-subjects... It also allows the focus to be on 

aerosol application only as opposed to the broad range of janitorial activities a subject 
Attachment 1 

Page 20 of 52 

might engage in...” (V1:18). Finally, the geographic area selected in close to the 

laboratory conducting the study and will save resources. 

Amount of AI Handled. The amount of active ingredient handled will be varied 

among monitoring events by varying the number of cans applied. The AEATF II 

cites existing survey data the Antimicrobial Exposure Joint Venture (AEJV) and 

information provided by JohnsonDiversey which was based on was based in part on 

information from the following sources: 1) the American Hospital Association 

(http://www.aha.org/aha_app/index.jsp), the American Society for Health Care 

Environmental Services (http://www.ashes.org/ashes_app/index.jsp), and the U.S. 

EPA’s Environmental Best Practices for Health Care Facilities. The AEJV indicates 

that consumers spray an average of ~113 g product per room (kitchen or bathroom) 

(or ~20% of a 19 oz can). Information from JohnsonDiversey indicates that a single 

individual would clean 20 rooms/day, which is considered to be a reasonable upper 

bound value. Therefore, the maximum amount of total product applied daily by a 

professional janitor is 2,260 g (or 4.2 19-oz cans). The aerosol protocol will monitor 

subjects spraying from 1 to 4 19-oz aerosol cans. (V1:22) EPA/CDPR/PMRA 

concurs that the four aerosol cans is an appropriate maximum rate. 

Varying Subjects. The goal of the study is to monitor professional applicators, so 

the selection of janitors is appropriate to the goal of the study. Each monitoring event 

will use a separate individual to maximize diversity. “Each selected worker provides 

his/her unique set of behaviors to the aerosol application task...this study permits 

only one monitoring event per worker in order to capture a larger diversity of 

application behaviors” (V1:17). The use of professional applicators is believed to be 

a reasonable surrogate for the range of potential users that may apply aerosol 

antimicrobial products, including consumers, school janitors, hospital janitors, 

commercial lodging janitors, employees of janitorial services firms, etc. 

(b) How have random elements been incorporated into the scenario sampling design? 

“Surrogate worker selection: This process results in a simple random sample of 

qualifying subjects from the volunteer pool. Note, however, that this is not technically 

the same as a random sample from the existing population of professional janitorial 

workers. By definition, volunteers are self-selected and could, in theory, have different 

characteristics than non-volunteers. Such fine distinctions have little relevance in this 

case, however, because this is not an observational study of existing applicator-days. 

Because workers are randomly assigned to synthetic application-day conditions, the 

resulting MEs are still considered synthetic applicator-days. Thus, any type of random 

sampling of just one ME component (e.g., applicator in this case) provides no statistical 

advantage other than reduction of selection bias.” (V1:17) 

A stratified random sampling approach will be used to locate acceptable facilities. 

(V1:19) 
Attachment 1 

Page 21 of 52 

“This list of commercial lodging facilities will then be randomized. Next, these properties 

will be investigated in (random) order until NC qualifying facilities have been found.” 

(V1:19) 

“Individuals who are enrolled to participate in the study will then be randomly ordered 

and assigned a subject identification sequence number (SISN). ... This process results in 

a simple random sample of qualifying subjects from the volunteer pool.” (V1:17) 

“...workers are randomly assigned to synthetic application-day conditions...” (V1:17) 

“The N=18 randomly sampled professional applicators will be randomly assigned to the 

N=18 combinations of building/room configuration category and application volume. 

This provides a diverse set of MEs with respect to three meta-parameters: (1) applicator, 

(2) types of rooms, and (3) product volume applied. ” (V1:31) 

(c) What feasible opportunities to incorporate random elements in the design—if any— 

have been overlooked? 

No feasible opportunities to incorporate additional random elements in the sampling 

design are apparent. 

(d) What typical patterns of exposure will likely be included by the sampling design? 

“The aerosol application scenario is defined as the hand-held pressurized aerosol-based 

application of a label-specified end-use formulation containing an antimicrobial 

chemical. This includes the task of actual aerosol spraying for purposes of air and surface 

odor elimination, sanitizing, or disinfecting. The aerosol application scenario involves 

application according to typical practices, e.g., spraying surfaces from a distance of 

approximately 6-10 inches in a manner to apply enough formulation to provide an 

adequate amount for cleaning. Hard surface applications are typically sprayed until 

visibly “thoroughly” wet per label direction. ... Surface applications are typically made in 

smooth, sweeping, overlapping patterns. Examples of “representative” spray application 

techniques that this scenario is expected to capture include horizontal spraying moving 

upward and downward from the starting point to hard surfaces such as laminate, tile, 

porcelain, glass, and metal).” (V1:8) 

Application surfaces include horizontal and vertical surfaces, kitchens, bathrooms, sinks, 

countertops, toilets, (V1:19) and food preparation areas (or FPA) which is defined as a 

room containing a stove/oven, refrigerator, and food preparation sink (V1:30). 

“The Clorox Disinfecting Spray will be applied by janitorial professionals according to 

typical practices, i.e., spraying surfaces from a distance of approximately 6-10 inches in a 

manner to apply enough formulation to provide an adequate amount for cleaning. Hard 

surface applications are typically sprayed until visibly ‘thoroughly’ wet per label 

direction. No wiping will be conducted as part of this application scenario. Surface 

applications are typically made in smooth, sweeping, overlapping patterns. Examples of 

‘representative’ spray application methods that this study is expected to capture (i.e., 
Attachment 1 

Page 22 of 52 

“horizontal spraying moving upward and downward from the starting point to hard 

surfaces such as laminate, tile, porcelain, glass, and metal”) are specified in the informed 

consent form.” (V2:21) 

Additional sources of aerosol application exposure include “spraying in an enclosed 

space (e.g., shower enclosure), spraying above and below the chest height, spraying near 

air exhaust vents, or walking into spray mist sprayed overhead.” (V1:22) 

(e) What typical patterns of exposure will likely be excluded by the sampling design? 

The study will monitor only professional janitors, so any differences in behavior that a 

consumer may exhibit while applying an aerosol product will not be monitored. 

However, given that spraying a pressurized can is not a specialized task, the exclusion of 

consumers from the sampling design is not considered to be a limiting factor. 

The aerosol scenario excludes the task of post-application wiping. The exposures to 

wiping a liquid film will be monitored in a separate wiping study as outlined in the 

AEATF II Governing Document. “In practice, aerosol application may or may not 

involve follow-on tasks, such as wiping the sprayed surface. Some aerosol products are 

considered ‘leave-on’ sprays that do not require post-spray wiping. Only the actual 

aerosol spray application is covered by this scenario. Applicator exposure associated with 

wiping is being addressed in another AEATF II scenario. Therefore, in the case of the 

aerosol application scenario, the applicator’s exposure during a single workday would 

arise only from the task of application (spraying) of the product (i.e., not from postapplication wiping, such as a potable water rinse and wipe, following aerosol application 

to a food preparation surface).” (V1:9) 

3. Is the proposed test material an appropriate surrogate? 

The test substance, Clorox Disinfecting Spray, is an appropriate choice for the 

development of surrogate exposure data because it is stable, is characterized by a low 

vapor pressure (estimated to be 2E-11 mmHg), diluted solutions are not dermally 

irritating, no systemic toxicity was seen in a 90-day dermal rat study, the product is 

registered for aerosol sprays, and its principal active ingredient ADBAC has been 

demonstrated to have a low analytical limit of detection. 

“The test substance for these studies is the formulated product, Clorox Commercial 

Solutions® Clorox® Disinfecting Spray (referred to as Clorox Disinfecting Spray in this 

protocol), containing didecyl dimethyl ammonium chloride (DDAC), n-Alkyl (C12, C14, 

and C16) dimethyl benzyl ammonium chlorides (ADBAC), octyl decyl dimethyl 

ammonium chloride (ODAC), and dioctyl dimethyl ammonium chloride (DODAC). The 

quaternary ammonium antimicrobials are commonly known as “quats”. C14 ADBAC is 

the active ingredient selected for measurement, based on its stability, abundance in the 

formulation, and sensitivity of its analytical method.” (V2:18) 
Attachment 1 

Page 23 of 52 

“Clorox Disinfecting Spray is an end use product registered with the EPA for use on 

smooth surfaces in indoor environments. Clorox Disinfecting Spray contains ADBAC, 

DDAC, ODAC and DODAC. ADBAC was selected as the analyte based primarily upon 

its abundance in aerosol products, on its stability, and the sensitivity of its analytical 

method. The quats ADBAC, ODAC, DODAC, and DDAC have complete toxicology 

databases with low mammalian toxicity. Virtually all quat antimicrobial products contain 

more than a single quat, i.e., a readily available product containing only ADBAC was not 

apparent. The analytical method for ADBAC on the proposed monitoring matrices at 

very low concentrations has been validated (GPL, 2004). The freezer storage stability of 

ADBAC on the different matrices to be used in this study is ongoing (GPL, 2009; in 

progress).” (V2:19) 

“The very sensitive and selective analytical method developed for the analysis of 

ADBAC on different study matrices will allow for the detection and quantification of 

extremely low levels of active ingredient in the collected samples. This will allow for 

shorter exposure time, thus minimizing the risk to research study subjects. Additionally, 

Clorox Disinfecting Spray has been deemed suitable by the Sponsor and EPA as a 

surrogate compound for generating exposure data for other antimicrobial pesticides.” 

(V2:20) 

ADBAC and DDAC have been approved for use in many formulations, and are 

extensively used in many janitorial products. The EPA has recently re-registered both 

DDAC and ADBAC and issued REDs for both (EPA, 2006a,b). Additionally, the safety 

of the test material has been established through long term professional use of the 

product. The product will be used according to its label. 

4. What is the rationale for the proposed cluster design and sample size? 

AEATF II proposes for the aerosol scenario the design of 3 clusters of 6 monitoring 

events (MEs) each. This configuration is appropriate to the characteristics of this 

scenario. 

“The primary challenge is that for the aerosol application scenario (as is true for all 

AEATF II scenarios) only a small number of expensive experimentally-obtained 

monitoring events are feasible.” (V1:14) 

“For the aerosol application study, therefore, random nested sampling will be used as a 

reasonable reference model for the combination of random sampling, randomization, and 

diversity selection actually used.” (V1:24) 

“For a configuration of NC=3 clusters (i.e., sites or buildings), Table 6 shows the sample 

size necessary to achieve 3-fold relative accuracy with GSD=2.1 or GSD=2.9 and an 

intra-cluster correlations (ICC) as high as 0.3. For the aerosol-only GSD of 2.1, only 2 

MEs per cluster are needed giving a total of N=6 MEs for the aerosol scenario. However, 

when the more robust repetitive-motion task GSD of 2.9 is used, 6 MEs per cluster are 
Attachment 1 

Page 24 of 52 

required giving N=18 MEs for the scenario. Given the sensitivity of the sample size to 

GSD and the belief that the repetitive-motion GSD is a better indicator of the expected 

true relative variation for this scenario, the AEATF II prefers to assume GSD=2.9 for the 

purposes of determining sample size. As also shown in Table 6, smaller, and perhaps 

more likely, ICCs will yield accuracies much better than 3-fold.” (V1:29) The reader is 

referred to Table 6 in Volume 1 (page 29) for a review of the results of the accuracy 

given n=2 versus n=6 MEs per cluster. 

“Benchmark objectives specify accuracy goals that must be achieved within the 

framework of the reference sampling model when sample size is adequate. In this study, 

‘sample size’ means both the number of clusters (NC) and the number of MEs per cluster 

(NM). For the aerosol application study, the benchmark objective is that (when the 

reference model is true) sample estimates of the arithmetic mean and 95th percentile of 

normalized exposure are accurate to within 3-fold 95% of the time.” (V1:24) 
Attachment 2 

Page 25 of 52 

EPA Protocol Review: AEATF II Aerosol Application Scenario/Protocol 

Title: A Study For Measurement of Potential Dermal and Inhalation Exposure 

During Application of a Liquid Antimicrobial Pesticide Product Using a 

Pressurized Aerosol Can for Indoor Surface Disinfecting. 

Date: August 4, 2009 

Principal Investigator: 

 Sami Selim, Ph.D. 

 

Participating Laboratories: 

 Golden Pacific Laboratories, LLC 

 4720 W. Jennifer Ave. Suite 105 

 Fresno, CA 93722 

 Eurofin | Grayson 

 211 N Main Street 

 Creedmoor, NC 27522 

Sponsor: American Chemistry Council 

Antimicrobial Exposure Assessment Task Force II 

c/o Hasmukh Shah, Ph.D. 

1300 Wilson Blvd 

Arlington VA 22209 

 

Reviewing 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 primary purpose of the aerosol application monitoring study is to develop more 

accurate information on worker exposures to antimicrobials . . . that broadly represent 

those expected for the future application of arbitrary antimicrobial pesticides.” (V1:13) 

 

“The main goal of the study is to generate data using a product with high exposure 

potential and covering the most influential variables associated with inhalation and 

dermal exposure. The data generated can then be used in risk assessments for most 

exposure scenarios resulting from use of [antimicrobial products in] aerosol cans.” 

(V1:38) 
Attachment 2 

Page 26 of 52 

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

Would the research fill an important gap in understanding? 

“This study is being conducted to determine potential dermal and inhalation antimicrobial 

chemical exposures associated with the use of hand-held, pressurized aerosol cans.” 

(V1:8) 

“Since 1992 the EPA has conducted professional and consumer mixer/loader and 

applicator exposure and risk assessments relying primarily on the exposure data in 

PHED. . . . PHED does include two studies conducted using aerosol application 

methods... Unfortunately, these studies have limitations that reduce their value for an 

antimicrobial-oriented generic database.” (V1:10-11) 

“In addition to PHED, another source of existing data being used by regulatory agencies 

in the case of antimicrobials is the Chemical Manufacturers Association Antimicrobial 

Exposure Assessment Study directed by Dr. William Popendorf at the University of Iowa 

(Popendorf et al. 1992)... The EPA concluded that the limited number of replicates 

combined with poor recovery data severely limits the conclusions that can be made from 

the CMA study. Therefore, the Agency is requiring confirmatory data to support the uses 

assessed with the CMA exposure data within this risk assessment. The limitations 

identified by EPA in the CMA study data were also echoed by regulatory agencies in 

California and Canada. All note that the exposure data cannot be used as generic data for 

all antimicrobials because recoveries were low, precision of the measurements were not 

established, and CMA did not establish the validity of generalizing the information 

among applications and end-use settings.” (V1:12-13) 

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

EPA will consider the data from this study in assessing exposures of occupational or 

residential applicators using hand-held, pressurized aerosol cans to apply an antimicrobial 

pesticide to indoor surfaces. 

 

(d) Could the research question be answered with existing data? If so, how? 

Due to the limitations of existing data, the research question cannot be answered with 

confidence relying on existing data. 

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

how? If not, why not? 

“Human subjects are required in this study because they will normally be exposed to the 

test material when performing their daily activities. There are no acceptable methods or 

models that could be used to extrapolate subjects’ exposure.” (V2:13) 
Attachment 2 

Page 27 of 52 

2. Study Design 

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

is it? 

“The primary purpose of the aerosol application monitoring study is to develop more 

accurate information on worker exposures to antimicrobials. These data will consist of 

dermal and inhalation exposure estimates derived from monitoring subjects under 

conditions that broadly represent those expected for the future application of arbitrary 

antimicrobial pesticides. 

Although this study will only monitor a single active ingredient, AEATF II and 

regulatory agencies generally recognize two important principles that allow such 

exposure results to be generalized to a larger set of conditions: 

1. Dermal and inhalation exposure to antimicrobial chemicals are considered generic 

(i.e., independent of the particular active ingredient used). This generic principle 

permits use of a single surrogate active ingredient to predict exposure for other 

active ingredients. 

2. The principle of proportionality of exposure to appropriate measures of active 

ingredient contact potential. For example, if measured exposure is E1 when the 

amount of active ingredient handled (AaiH) is H1, then the predicted exposure 

when AaiH is H2 is just E2 = H2(E1/H1). 

Consequently, AEATF II anticipates the resulting database will contain sufficient data to 

support exposure assessments for aerosol application for a number of antimicrobial active 

ingredients over a range of AaiH levels.” (V1:13) 

“Benchmark objectives specify accuracy goals that must be achieved within the 

framework of the reference sampling model when sample size is adequate. In this study, 

‘sample size’ means both the number of clusters (NC) and the number of MEs per cluster 

(NM). 

For the aerosol application study, the benchmark objective is that (when the reference 

model is true) sample estimates of the arithmetic mean and 95th percentile of normalized 

exposure are accurate to within 3-fold 95% of the time. The EPA, in discussion with 

AEATF II, determined that this benchmark is sufficient for regulatory purposes.” (V1:24) 

No hypothesis is stated, nor is the study designed to test a hypothesis. 

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

The objective cited above can be achieved by the study as proposed. 
Attachment 2 

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2.1 Statistical Design 

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

The goal is to use these data to characterize some ‘population’ aspect of the future 

exposure to arbitrary antimicrobial pesticides. Hence, this study is more closely 

aligned with the random sampling situation. The “...random reference model 

approach is used to determine sample sizes for the aerosol application scenario. The 

aerosol application study will utilize a combination of random sampling, 

randomization, and diversity selection methods. While this methodology contains 

some elements of all three pure situations ....[statistical theory], none apply 

completely. The ultimate goal of this study is to construct synthetic MEs that can be 

used to characterize the diversity of future daily exposures to antimicrobials through 

aerosol application. Hence, the study objectives are more closely aligned with the 

random sampling situation.... As a result, a reference model for random sampling 

will be used for the determination of sample size.” (V1:23) 

“For the aerosol application study, therefore, random nested sampling will be used as 

a reasonable reference model for the combination of random sampling, 

randomization, and diversity selection actually used. This reference model assumes 

that: 

1. Exposure, normalized by the amount of active ingredient handled, is 

lognormally distributed with a known geometric standard deviation (GSD). 

Equivalently, the logarithm of normalized exposure is normally distributed 

with known standard deviation Log GSD. 

2. There are NC clusters (i.e., sites) and NM MEs per cluster. The total number 

of MEs is, therefore, N=NC×NM. 

3. There is a possible within-cluster (i.e., within-site) correlation of log 

normalized exposure. This is referred to as the intra-cluster correlation, or just 

the ICC. “ (V1:24) 

“...the benchmark objective is that (when the reference model is true) sample 

estimates of the arithmetic mean and 95th percentile of normalized exposure are 

accurate to within 3-fold 95% of the time.” (V1:24). 

“A Monte Carlo simulation approach was used to examine the impact of number of 

clusters (NC) and number of MEs per cluster (NM) on accuracy of the arithmetic 

mean and 95th percentile for the reference model.” (V1:28). 

“For the aerosol-only GSD of 2.1, only 2 MEs per cluster are needed giving a total of 

N=6 MEs for the aerosol scenario. However, when the more robust repetitive-motion 

task GSD of 2.9 is used, 6 MEs per cluster are required giving N=18 MEs for the 

scenario. Given the sensitivity of the sample size to GSD and the belief that the 

repetitive-motion GSD is a better indicator of the expected true relative variation for 
Attachment 2 

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this scenario, the AEATF II prefers to assume GSD=2.9 for the purposes of 

determining sample size.” (V1:29) 

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

appropriate for the study design and statistical analysis plan? 

No positive or negative controls are proposed. This is appropriate for the study 

design and statistical analysis plan. 

(c) How is the study blinded? 

The study is not blinded. 

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

Candidates selected from among respondents to recruiting flyers and/or newspaper 

advertisements will be assigned randomly to one or another ME, pre-defined in terms 

of its amount handled and location. (V2:26) 

MEs with more aerosol cans to be sprayed will be executed first; a subject who is 

unable to continue spraying for the given amount will be reassigned to a lesser 

amount ME, and the next subject in random order will be assigned to the 

uncompleted ME. (V2:26) 

“As long as each subject achieves the target spraying amount, the process is 

continued down to the smallest spray amount stratum (A, 1 to 1.5 canisters) and six 

MEs have been obtained, one for each of the six strata. If this process proceeds as 

expected, the last two subjects allocated to each site are never used for MEs and will 

remain alternates.” (V2:26) 

(e) Can the data be statistically analyzed? 

The results of the analysis from the sampling will be provided in the final report. 

(V2:51, 52). 

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

“The AEATF II will not statistically analyze the monitoring data in order to 

characterize exposure or investigate the relationship between exposure and other 

factors (e.g., room size, level of residual organic matter, environmental conditions 

including temperature, humidity, air turnover rate, etc.) However, regulators and 

other users of the constructed database (BHED) may choose to conduct such 

analyses. The extent of AEATF II’s data analyses will be limited to the statistical 

characterization of data adequacy for inclusion in BHED scenario monographs. Two 

specific types of analyses will be performed (these analyses are discussed in more 

detail in the AEATF II’s Governing Document (AEATF II, 2008). 
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1. Evaluation of benchmark adequacy. A confidence interval based approach 

will be used to determine the realized relative accuracy for the arithmetic 

mean and 95th percentile of exposure normalized by amount of ai handled. 

2. Cluster effects. The intraclass correlation for clusters (ICC) and its confidence 

interval will be estimated using a variance components model. In addition, the 

effects, if any, of ignoring clusters in the estimation of means and percentiles 

will be determined by comparing the estimates of a no-cluster model to those 

of the random effects model.” (V2: 47, 48) 

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

 Yes. 

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

the research question? 

Because of its Purposive Diversity Sampling Design, the study will support only 

limited inferences. EPA believes, nonetheless, that it is likely to characterize reliably 

the exposures of professional janitors in the Fresno area, that those exposures are 

likely to be similar for other professional janitors elsewhere, and that the exposures of 

professional janitors spraying pressurized aerosol cans for extended durations can 

inform assessments of the likely exposure of others spraying aerosol products for 

shorter durations with other types of aerosol products/cans. EPA is confident that this 

design will provide data on aerosol spray exposures more accurate and reliable than 

currently available data. 

2.2 How and to what will human subjects be exposed? 

The test subjects will be exposed to Clorox Commercial Solutions® Clorox® 

Disinfecting Spray (EPA Reg. No. 67619-03), which contains active ingredients 0.0945% 

didecyl dimethyl ammonium chloride (DDAC), 0.252% n-Alkyl (40% C12, 50% C14, 

and 10% C16) dimethyl benzyl ammonium chlorides (ADBAC), 0.189% octyl decyl 

dimethyl ammonium chloride (ODAC), and 0.0945% dioctyl dimethyl ammonium 

chloride (DODAC). (V1:9; V2:19). 

Test subjects will be exposed during aerosol spraying activities using Clorox Commercial 

Solutions® Clorox® Disinfecting Spray (EPA Reg. No. 67619-03). The aerosol 

application scenario involves application according to typical practices, e.g., spraying 

surfaces from a distance of approximately 6-10 inches in a manner to apply enough 

formulation to provide an adequate amount for cleaning. Hard surface applications are 

typically sprayed until visibly “thoroughly” wet per label direction. No wiping will be 

conducted as part of this application scenario. Surface applications are typically made in 

smooth, sweeping, overlapping patterns. Examples of “representative” spray application 

techniques that this scenario is expected to capture include horizontal spraying moving 
Attachment 2 

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upward and downward from the starting point to hard surfaces such as laminate, tile, 

porcelain, glass, and metal. (V1:8) 

“Each ME will apply the test substance to bathrooms and/or food preparation areas which 

include horizontal and vertical surfaces (e.g. shower stall/tub, toilet, countertops, sinks, 

cabinets and appliances). Each bathroom is expected to include some combination of 

shower stall/tub enclosure, toilet, and/or sink/countertops. Each kitchen or food 

preparation area is expected to include counter tops and some appliance surfaces. All 

interior surfaces of the shower/tub, the horizontal surface of the countertop/sink, and 

exterior surface only of toilets will be sprayed with the test substance. One complete can 

of aerosol product will cover the shower, sink and toilet in approximately two bathrooms 

if every surface were sprayed; however, that is not typical practice and each participant 

will be encouraged to apply the spray as they would in normal practice.” (V2:21) 

“The procedures described represent typical consumer and professional worker methods 

of applying the test substance to indoor surfaces. The test substance chosen is utilized in 

the described scenario that employs aerosol application of antimicrobials.” (V2:22) 

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

“Based on the nozzle size, amount of material dispensed per unit time, air 

concentrations, and aerosol characteristics, across products in the four major aerosol 

categories (hard surface disinfecting sprays, hard surface foaming aerosols, soft 

surface sprays, air fresheners), Clorox Commercial Solutions® Clorox® Disinfecting 

Spray (EPA Reg. No. 67619-03), was selected as a representative, albeit conservative 

surrogate and a product most likely to produce measurable exposures for purposes of 

the aerosol application study.” (V1: 9) 

“C14 ADBAC is the active ingredient selected for measurement, based on its 

stability, abundance in the formulation, and sensitivity of its analytical method” 

(V2:18) 

“Clorox Disinfecting Spray is an end use product registered with the EPA for use on 

smooth surfaces in indoor environments. Clorox Disinfecting Spray contains 

ADBAC, DDAC, ODAC and DODAC. ADBAC was selected as the analyte based 

primarily upon its abundance in aerosol products, on its stability, and the sensitivity 

of its analytical method. The quats ADBAC, ODAC, DODAC, and DDAC have 

complete toxicology databases with low mammalian toxicity. Virtually all quat 

antimicrobial products contain more than a single quat, i.e., a readily available 

product containing only ADBAC was not apparent.” (V2:19) 

“The analytical method for ADBAC on the proposed monitoring matrices at very low 

concentrations has been validated (GPL, 2004). The freezer storage stability of 

ADBAC on the different matrices to be used in this study is ongoing (GPL, 2009; in 

progress).” (V2:19) 
Attachment 2 

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“The very sensitive and selective analytical method developed for the analysis of 

ADBAC on different study matrices will allow for the detection and quantification of 

extremely low levels of active ingredient in the collected samples. This will allow for 

shorter exposure time, thus minimizing the risk to research study subjects. 

Additionally, Clorox Disinfecting Spray has been deemed suitable by the Sponsor and 

EPA as a surrogate compound for generating exposure data for other antimicrobial 

pesticides.” (V2:20) 

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

dose administration? 

In this study all MEs will apply the same substance, i.e., Clorox Disinfecting Spray in 

19 oz (538 g) canisters. This substance contains several active ingredients. However, 

only one of these, C14 ADBAC, will be quantified. Because the concentration of 

ADBAC in the aerosol canisters is the same for all MEs, the amount of active 

ingredient ‘handled’ is varied simply by having MEs that apply different amounts of 

the formulated product. The total amount of product sprayed, will range between 1 

and 4 canisters (i.e., 538 to 2152 grams of formulated product). (V2:25) 

ADBAC was selected because of its relatively high concentration, its stability, and 

the sensitivity of the analytical method. 

The same concentration will be used by all MEs. The total amount of ai handled will 

vary with the number of aerosol cans sprayed. 

(c) What duration of exposure is proposed? 

Each predefined ME will apply different amounts of the formulated product. 

Therefore, the amount of product applied is proposed, not specific time durations. 

The AEATF II does anticipate that the duration to apply the specified amounts will 

range from 30 to 180 minutes. The total amount of product sprayed, will range 

between 1 and 4 canisters (i.e., 538 to 2152 grams of formulated product). This 

interval is partitioned into six amounts handled for aerosol spray strata as follows: 

A. 1 to 1.5 canisters 

B. 1.5 to 2 canisters 

C. 2 to 2.5 canisters 

D. 2.5 to 3 canisters 

E. 3 to 3.5 canisters 

F. 3.5 to 4 canisters 

Within each monitoring site, a single ME is targeted for each of the above six strata. 

As long as the subject is within the correct AaiH interval, it is unnecessary to control 

the exact amount of product precisely. However, the actual amount of product 

sprayed by each subject will be recorded. (V2:25) 
Attachment 2 

Page 33 of 52 

Any test subjects not able to spray the given number of canisters are allowed to stop 

and as long as at least one canister is applied the dosimeters will be analyzed. If the 

subjects fail to spray a single canister, the ME will not be used and the monitoring 

media will not be analyzed. (V2:26) 

The AEATF II cites existing survey data the Antimicrobial Exposure Joint Venture 

(AEJV) and information provided by JohnsonDiversey which was based on was 

based in part on information from the following sources: 1) the American Hospital 

Association (http://www.aha.org/aha_app/index.jsp), the American Society for Health 

Care Environmental Services (http://www.ashes.org/ashes_app/index.jsp), and the 

U.S. EPA’s Environmental Best Practices for Health Care Facilities. The AEJV 

indicates that consumer’s sprays an average of ~113 g product per room (kitchen or 

bathroom) (or ~20% of a 19 oz can). Information from JohnsonDiversey indicates 

that a single individual would clean 20 rooms/day, which is considered to be a 

reasonable upper bound value. Therefore, the maximum amount of total product 

applied daily by a professional janitor is 2,260 g (or 4.2 19-oz cans). The aerosol 

protocol will monitor subjects spraying from 1 to 4 19-oz aerosol cans. (V1:22) 

2.3 Endpoints and Measures 

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

being asked? 

“Potential dermal exposure to the test substance will be measured externally using 

whole body inner and outer dosimeters, hand washes, and face/neck wipes. All 

monitored subjects will wear the outer dosimeter (representative outer clothing 

consisting of cotton long pants and cotton long sleeve shirts) directly over the inner 

dosimeter (consisting of 100% cotton long underwear). Inner and outer dosimeters 

will be provided by AEATF. Subjects will wear their own socks and shoes. Hand 

exposure will be measured by rinsing the hands with a solution of 50% isopropyl 

alcohol/ 50% distilled water. Face and neck exposure will be measured by wiping the 

face and neck with gauze pads moistened with 50% isopropyl alcohol / 50% distilled 

water.” (V2:11) 

“The potential total inhalation exposure for each subject will be measured with an 

OSHA Versatile Sampler (OVS) tube attached to a personal air sampling pump set at 

a typical sampling rate (2 L/minute). Potential exposure to respirable, thoracic and 

inhalable particles (100, 10 and 2.5 μm, respectively) will be characterized with a 

three stage RespiConTM Particle Sampler (Model 8522, TSI Inc.) attached to a 

personal air sampling pump operating at ~3.1 L/min.” (V2:12) In addition, presampling for ambient air will be performed to determine if there are background 

ADBAC air concentrations. (V2:34) 

“Airflow of pumps attached to the OVS tube and RespiConTM sampler will be 

calibrated to a nominal flow rate of approximately 2 and 3.1 L/min respectively using 
Attachment 2 

Page 34 of 52 

SOPs of field study procedures. The beginning flow rate of each pump will be 

checked and documented just before being fitted to the subject.” (V2:35, 36) 

“The inner and outer dosimeters, OVS tubes, filters from the RespiConTM Particle 

Sampler, hand wash solutions, and face/neck wipes will be analyzed for residues of 

C14 ADBAC using validated analytical methods.” (V2:12) 

“The amount of test substance applied during each ME will be determined by the 

total change in weight of all canisters used in the ME. Each canister will be uniquely 

identified prior to experimental start. Prior to each ME a sufficient number of unused 

canisters will be individually weighed, and the beginning weights of each canister 

recorded in the raw data. Following completion of the ME, each canister used in the 

ME will be individually weighed, and the end weight and amount of change for each 

canister recorded in the raw data. The total amount of test substance applied will be 

determined by the amount of change (i.e., test substance discharged) from all 

canisters used in the ME. All information necessary to reconstruct the amount of test 

substance applied by each ME will be documented.” (V2:22) 

“Air temperature and relative humidity of the work area for the duration of exposure 

monitoring will be documented with automated instrumentation logging and 

recording at intervals appropriate for the duration of the work period per SOP 

AEATF II-10C. Environmental monitoring equipment will be calibrated or 

standardized according to SOPs. HVAC and room volume will be described in detail 

and documented in study field notes, and it will be noted for each room sprayed 

whether the HVAC and/or vent fans were on at the time of application.” (V2:40) 

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

The AEATF II SOPs provide specific procedures to ensure accurate measurements 

such as calibration of inhalation monitoring devices (SOP 10G.1 (V4:107)). 

“GLP purity analysis (content of active ingredient in the test substance) will 

be performed by the Sponsor, and a Certificate of Analysis will be kept in 

the raw data file.” (V2:19) 

“Full details regarding field recovery evaluation procedures for all sampling 

media are given in the most recent version of the SOPs of the AEATF II-8E[.1].” 

(V2:41)

(c) What QA methods are proposed? 

Volume 2, Section 12, Analytical Procedures, discusses QA methods for reference 

substances, fortification solutions, internal standards, analytical method, etc. (V2:44-

48) 
Attachment 2 

Page 35 of 52 

Volume 2, Section 10.A, Air Sampling for Ambient Pre-existing ADBAC, discusses 

sampling for background air concentrations of ADBAC (V2:34) 

“This study will be conducted according to FIFRA GLP Standards (40 CFR 160).” 

(V2:51) 

(d) How will uncertainty be addressed? 

In a September 18, 2009, letter from Hasmukh Shah (Manager, AEATF II) to 

Timothy Leighton (Office of Pesticide Programs, Antimicrobial Division), the 

AEATF II committed to address deviations from the benchmark accuracy goal (i.e., 

k=3) as follows: “[I]f large deviations from the benchmark goals are observed in the 

aerosol study, AEATF II will, in consultation with regulatory agencies, determine the 

best course of action to take. This may mean the development of guidance for the use 

of these data that takes the increased imprecision of the estimates into account. It is 

possible that collection of additional clusters might be considered.” (a copy of the 

letter is included in the supplemental materials). 

 

3. Subject Selection 

3.1 Representativeness of Sample 

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

The target population is the distribution of future handler/days of applicators applying 

antimicrobial pesticides in the aerosol scenario. 

“The community of individuals that use an aerosol to clean/disinfect surfaces is 

enormous and includes millions of workers and at least 100 million residents in the 

U.S. alone” (V2:17-18) 

(b) From what populations will subjects be recruited? 

“Adult subjects will be recruited from the janitorial/cleaning service population of 

Fresno County. The most-recent US Census indicates that 40% of the population in 

Fresno, CA metropolitan area is Hispanic. The proportion of Hispanics in service 

industries, e.g., janitorial services, may be even higher than the general population. 

Therefore to adequately capture the ethnic diversity in the Fresno area, recruitment 

materials and all communications with potential subjects will be available in English 

and Spanish, as preferred by the subject.” (V2:27) 

“IRB approved recruiting advertisements...will be simultaneously (within the same 

week) placed in the Fresno Bee, the California Advocate and the Fresno edition of 

Vida en el Valle. The Fresno Bee is a large, general circulation daily paper in Fresno 

County. The California Advocate is the dominant African American community 
Attachment 2 

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weekly paper in Fresno County, and Vida en el Valle is a weekly targeting the San 

Joaquin Valley, with separate editions for Fresno and other central valley 

municipalities.” (V2:28) 

(c) Are expected participants representative of the population of concern? If not, 

why not? 

Expected participants will self-identify in response to flyers soliciting interest, posted 

in the workplaces of janitorial services companies, or in response to advertisements 

placed in three Fresno-area newspapers. Volunteers may differ in unknowable ways 

from other workers who do not step forward. There is no reason to think that 

janitorial service workers in Fresno are atypical of janitorial service workers in any 

other area of the United States. 

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

sample? 

“The AEATF II program, as described in the Governing Document (2008), intends to 

develop a database of exposure monitoring data that can be used to support practical 

regulatory decisions about future exposures for different (including currently 

nonexistent) active ingredients and their associated products. The database needs to 

address a variety of exposure scenarios for which no or limited data currently exist. 

The aerosol application scenario is an important component of the AEATF II program 

and the focus of this study. As noted in the previous section, existing monitoring data 

for this scenario are considered inadequate. 

“The primary purpose of the aerosol application monitoring study is to develop more 

accurate information on worker exposures to antimicrobials. These data will consist 

of dermal and inhalation exposure estimates derived from monitoring subjects under 

conditions that broadly represent those expected for the future application of arbitrary 

antimicrobial pesticides. 

“Although this study will use only a single active ingredient, AEATF II and 

regulatory agencies generally recognize two important principles that allow such 

exposure results to be generalized to a larger set of conditions: 

1. Dermal and inhalation exposure to antimicrobial chemicals are considered 

generic (i.e., independent of the particular active ingredient used). This 

generic principle permits use of a single surrogate active ingredient to predict 

exposure for other active ingredients. 

2. The principle of proportionality of exposure to appropriate measures of active 

ingredient contact potential. For example, if measured exposure is E1 when the 

amount of active ingredient handled (AaiH) is H1, then the predicted exposure 

when AaiH is H2 is just E2 = H2(E1/H1). 
Attachment 2 

Page 37 of 52 

“Consequently, AEATF II anticipates the resulting database will contain sufficient 

data to support exposure assessments for aerosol application for a number of 

antimicrobial active ingredients over a range of AaiH levels. 

“An applicator-day is defined as a single professional applicator and a single day on 

which he/she performs the scenario-specific task as described in Section 2 above. 

Each possible applicator-day is implicitly associated with a set of application 

conditions that includes, but is not limited to, applicator behavior, formulation type, 

location, and environmental conditions. Therefore, the aerosol application scenario 

can be viewed as the collection (or ‘population’) of all possible applicator-days that 

conform to the scenario definition. The basic experimental unit for this scenario is a 

monitoring event (or ME). During a monitoring event, AEATF II researchers will 

collect dermal and inhalation exposure information from a worker while he/she 

performs aerosol application. Each ME is designed to represent a single applicatorday and its corresponding exposure potential. Therefore, the set of N MEs obtained 

for the aerosol application scenario are designed to characterize future aerosol 

application scenario applicator-days. The primary challenge is that for the aerosol 

application scenario (as is true for all AEATF II scenarios) only a small number of 

expensive experimentally-obtained monitoring events are feasible.” (V1:13-14) 

3.2 Equitable Selection of Subjects 

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

Inclusion/exclusion criteria are complete and appropriate. They are listed on V2:30-

31 and below: 

“Inclusion Criteria: 

• Males or females, at least 18 years of age 

• In good health 

• Willingness to sign the Informed Consent Form and Subject Self Reporting 

Demographic Form 

• Speak and read English or Spanish 

• Resident of Fresno County 

• Experience in providing janitorial services 

Exclusion Criteria: 

• Skin conditions on the surface of the hands (e.g., psoriasis, eczema, cuts or 

abrasions) 

• Pregnancy, as shown by a urine pregnancy test 

• Lactation 

• Allergies to household chemical-based products, soaps or isopropyl alcohol 

• Declines to sign the Informed Consent Form or the Subject Self Reporting 

Demographic Form 

• Does not read and understand English or Spanish 
Attachment 2 

Page 38 of 52 

• Is less than 18 years old 

• Is not in good health 

• Severe respiratory disorders (e.g., moderate or severe asthma, emphysema) 

• Cardiovascular disease (e.g., history of myocardial infarcts, stroke, congestive 

heart failure or uncontrolled high blood pressure) 

• Is an employee of, or is related by blood or marriage to an employee of 

Golden Pacific Laboratories, Grayson | Eurofins, or a cleaning product 

manufacturer.” 

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

Employees and relatives of employees of the investigators are excluded from 

participation as subjects. (V2:31) 

(c) If any potential subjects are from a vulnerable population, what is the 

justification for including them? 

No potential subjects are from a vulnerable population. 

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

The recruiting process is described in V2:27-30. 

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

specific safeguards are proposed to protect their rights and welfare? 

“Janitorial services providing professional cleaning services for commercial buildings 

in Fresno County, CA will be contacted by qualified research personnel and asked 

whether they would be willing to post a flyer soliciting volunteers for a study to be 

conducted independent of the janitorial service...If the firm wishes to post the flyer, 

research personnel will provide a general description of the study, explain the need 

and importance of remaining neutral (un-coercive) in their interactions with 

employees regarding study participation, and determine whether the flyer seems 

intelligible for the firm’s employees. Assuming the owner/manager still wishes to 

post the flyer, research personnel will provide approval to do so....To avoid the 

potential for coercion, subjects will not be recruited directly through contract 

janitorial service companies. Flyers will direct interested workers to contact research 

personnel directly.” (V2:27-28) 

3.3 Remuneration of Subjects 

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

“All individuals that show up for the informed consent interview will be compensated 

$20 in cash at completion of the interview for their time and inconvenience. All 

individuals who are qualified, sign the informed consent form, and report to their 
Attachment 2 

Page 39 of 52 

assigned study site, will receive $100 in cash for their time and inconvenience when 

they leave the study site, whether they are monitored or not. If a subject signs the 

consent form and fills out the demographic form, information that may disqualify 

them from participation may become evident. If this occurs, the disqualified subject 

will be paid $100 as if a participant and dismissed. The value for compensation is 

based roughly on a day’s wage of $100 and represents potential lost time from 

secondary sources of employment, travel time and incidental expenses incurred in 

study participation. Compensation will be provided to individuals who complete their 

assigned participation or who need to withdraw for whatever reason.” (V2:32) 

(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? 

Compensation will be in the form of cash (U.S. currency). “All individuals that show 

up for the informed consent interview will be compensated $20 in cash at completion 

of the interview...All individuals who are qualified, sign the informed consent form, 

and report to their assigned study site, will receive $100 in cash...when they leave the 

study site, whether they are monitored or not.” (V2:32) 

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? 

The proposed test material is EPA-registered, with an essentially complete supporting 

database. The test material is of low toxicity to mammals. 

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

Risks are of a reaction to the test material or the solvents used to obtain residues from 

hands and face/neck; of discomfort and possibly heat-related illness associated with 

wearing two layers of clothing; of discomfort or inconvenience from wearing the air 

sampling device; of embarrassment from disrobing in the presence of a research 

technician; of an unexpected result of pregnancy testing. (V2:14-17) 
Attachment 2 

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Risks discussed in the consent form include risk of a reaction to the test material, risk 

of discomfort, risk of stinging from alcohol wash and wipes, a small possibility of 

heat stress, risk of embarrassment, possible surprise at the results of a pregnancy test,. 

(V2:70-71) 

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

probability estimated? 

All identified risks are characterized as of low probability. No quantitative estimates 

are reported. 

4.2 Risk minimization 

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

Use of test materials shown to be of low toxicity to mammals; use in strict accord 

with approved labeling; exclusion of candidates known to be sensitive to quaternary 

ammonium compounds; exclusion of candidates in poor health or with broken skin on 

hands, face, or neck; testing in a controlled-temperature environment; alerting 

subjects to signs and symptoms of heat stress; monitoring heat index with associated 

stopping rules; limited time of exposure with rest periods at 30-min intervals, or more 

frequently if requested; close observation of subjects; training of experienced 

technicians to minimize embarrassment; incorporation of procedures to keep results 

of pregnancy testing private and to permit discrete withdrawal; provision of 

appropriate work clothing and PPE. (V2: 14-17, 20, 32-34) 

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

test materials? 

The formulated product for the aerosol study contains more than one active 

ingredient. Clorox Commercial Solutions Clorox Disinfecting Spray (EPA Reg. No. 

67619-03) contains the test substance, ADBAC, and another active ingredient (i.e., 

DDAC). Therefore, the established NOAELs for both ADBAC and DDAC are 

provided below. 

The ADBAC risk assessment developed to support the Reregistration Eligibility 

Decision (RED) document provides for the selection of the toxicological endpoints 

for risk assessment purposes. The dermal toxicological endpoints indicate no 

systemic toxicity. However, dermal irritation has been observed in 21- and 90-day 

dermal toxicity studies in guinea pigs and rats, respectively (MRIDs 41105801 and 

41499601, respectively). The short-term dermal endpoint selected from the 21-day 

study is 333 ug/cm2

 and 80 ug/cm2 in the 90-day study. The proposed use of 

ADBAC in this protocol by subjects exposed to 0.252% ADBAC in a formulated 

product will not trigger a risk of concern. The inhalation toxicological endpoint 

identified for ADBAC for all exposure durations is based on a developmental toxicity 
Attachment 2 

Page 41 of 52 

study in rabbits (MRID 42392801) with a NOAEL of 3 mg/kg/day. The inhalation 

risk assessment for aerosol inhalation exposure in the RED document indicates that at 

this low concentration of ADBAC no inhalation risks are anticipated. 

The DDAC risk assessment developed to support the Reregistration Eligibility 

Decision (RED) document provides for the selection of the toxicological endpoints 

for risk assessment purposes. The dermal toxicological endpoints indicate that low 

concentrations of DDAC (0.13% ai tested in a 21-day dermal toxicity study, MRID 

45656601) display no dermal irritation effects and no systemic effects up to and 

including the limit dose of 1000 mg/kg/day. The proposed use of DDAC in this 

protocol by subjects exposed to a concentration of 0.378% active ingredient (i.e., 

aggregate of octyl dimethyl ammonium chloride, dioctyl dimethyl ammonium 

chloride, and didecyl dimethyl ammonium chloride) will not trigger a risk of concern. 

The inhalation toxicological endpoint identified for DDAC for all exposure durations 

is based on two oral toxicity studies (prenatal developmental toxicity in rats, MRID 

41886701, and a chronic toxicity study in dogs, MRID 41970401). The selected 

NOAEL from both studies is 10 mg/kg/day. The inhalation risk assessment for 

aerosol inhalation exposure in the RED document indicates that at this low 

concentration of DDAC, no inhalation risks are anticipated. 

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

Heat stress index above 95 (V2:33) 

Other medical reasons (V2:32-33) 

“If two or more subjects withdraw or are withdrawn from the study for the same 

medical reasons, the study will be suspended until the cause of the withdrawal is fully 

investigated and determined.” (V2:33-34) 

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

injury to subjects? 

SOP 11.B.1 for Management of Heat Stress (V4:112-123) 

SOP 11.C.1 for Emergency Procedures (V4:124-126) 

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

“If a subject reports an adverse skin reaction during the work period, they will be 

asked to immediately stop working. Research staff will then assist the subject in 

gently washing exposed skin with clean water and mild soap. After drying the area 

with a clean towel, the Principal Investigator will be contacted for further 

instructions.

“The extra layer of clothing worn by subjects may increase the risk of heat-related 

illness. To minimize the possibility of heat stress, the study will be conducted indoors 
Attachment 2 

Page 42 of 52 

in an environment where the heat index (HI) is expected to be less than 85. Research 

personnel shall monitor the heat index, and stop subjects’ work if the heat index

exceeds 95. The SOP AEATF 11.B describes the procedure for identification and 

control of heat stress. The poster “Controlling Heat Stress Made Simple” will be 

posted in the subject dressing area, and the information contained on the poster 

available to subjects and research personnel at the field site.

“In brief, researchers will observe subjects for possible signs of early heat illness such 

as fatigue, dizziness, irritability, or decreased concentration, especially if the worker 

has been working for a while. If these symptoms are observed, the subjects will be 

asked whether they would like to rest for a moment. If they answer affirmatively,

they will stop working, be given their choice of water or a sports drink, and the 

Principal Investigator will be immediately contacted for further medical management 

instructions. If they answer negatively, they will be permitted to continue working, 

and frequently thereafter asked whether they would like to rest for a moment. Any

affirmative answer will be handled as described above.

“If subjects develop visible signs or report symptoms of distress such as pronounced 

fatigue, headache, cramps, feeling faint, increased pulse, muscle spasms, heavy 

sweating (or dry skin if previously sweating), extreme thirst, or rapid breathing, the 

subjects will be asked to stop working immediately, and given their choice of water

or a sports drink. The Principal Investigator will immediately be contacted for further 

medical management instructions. If the worker’s condition appears to be serious, a 

member of the study team will call 911 and allow emergency medical personnel to

respond and treat the subject.” (V2:33) 

(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? 

“Subjects will be instructed to inform the Principal Investigator or research staff 

immediately if they feel ill, suffer a skin reaction or experience any other 

unanticipated adverse effects they feel may be related to the study during or following 

conduct of the study.” (V2:32) 

“If two or more subject develop an adverse skin reaction after they leave the study 

site, all subjects will be contacted by the Principal Investigator to determine whether 

further medical management is appropriate.” (V2:33)

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

paid for? 

The informed consent form states: “If you are hurt while you are in this study, a 

nearby medical facility that knows about this study will provide care. If necessary, 

we will take you there. We will pay for needed medical treatment that is not paid for 

by your own insurance or by someone else. To find out more, or if you think you 
Attachment 2 

Page 43 of 52 

may have been hurt during the study, call Dr. Selim at Golden Pacific Laboratories 

(559-275-9091) from 9 am to 5 pm Monday through Friday.” (V2:71) 

To clarify the meaning of the phrase “by someone else” in the statement about 

compensation for research-related injuries, the AEATF II should make the following 

revision: 

o Current language: “We will pay for needed medical treatment that is not paid 

for by your own insurance or by someone else.” 

o Change to: “We will pay for needed medical treatment that is not paid for by 

your own insurance or by the insurance of a third party under which you are 

covered.” 

5. Benefits 

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

“While there are no direct benefits to the subjects participating in this research study, 

there are indirect benefits to both the volunteers and society. . . . If individual workers 

request their results, they may find that their work practice produces more or less 

exposure than average, and this could be a useful learning tool.” (V2:17) 

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

through the research? 

“Products containing antimicrobial chemicals are used extensively in hospitals, schools, 

homes, etc. to control pathogenic bacteria and viruses known to produce increased 

morbidity and mortality in humans, domestic animals and pets...Measuring exposure of 

workers in this research study will produce reliable data about the dermal and inhalation 

exposure of workers...performing these tasks. The resulting data will improve the 

completeness and accuracy of the database used by the EPA to assess exposure to these 

chemicals. The ability to accurately predict risk may allow other chemical classes of 

antimicrobials to also be registered based on exposure estimates generated from the data 

to be produced by this study.” (V2:17) 

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

research? 

“Results from the study may benefit EPA...by reducing uncertainty about the range of 

exposure experienced by consumers and workers handling antimicrobials. Registrants of 

antimicrobials will benefit because they will provide EPA with data on exposure that has 

been made a condition of re-registration for a number of antimicrobials, and they may be 

aided in registering new antimicrobials using the data generated from this study.” 

(V2:17)
Attachment 2 

Page 44 of 52 

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

The research is very likely to produce more accurate and reliable information concerning 

exposure in the aerosol scenario, with resulting societal benefits in the form of more 

accurate and confident assessments of applicator exposure and risk.

6. Risk/Benefit Balance: How do the risks to subjects weigh against the anticipated 

benefits of the research, to subjects or to society? 

The likely benefit to janitorial workers as a whole and to society in general, in the form of 

more accurate measurements of potential exposure to antimicrobial products, must be 

weighed against the risks to study participants. Aerosol antimicrobial products are 

widely used both by janitorial workers and the general public. Exposure data for this 

scenario meeting contemporary standards of reliability and quality will likely provide a 

significant benefit to society. Because the margins of exposure are acceptable for the 

product proposed for use in this research study, subjects are very unlikely to experience 

toxic effects, and because procedures will be in place to minimize these and other risks to 

participants, the likelihood of serious adverse effects is very small. In summary, the risks 

to study participants from participating in this study are reasonable in light of the likely 

benefit to society of the knowledge to be gained. 

7. Independent Ethics Review 

(a) What IRB reviewed the proposed research? 

Independent Investigational Review Board, Inc., Plantation FL (IIRB) 

(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? 

No. 

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

No. 
Attachment 2 

Page 45 of 52 

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

Yes. 

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

“This is a protocol for third-party research involving what EPA has interpreted to be 

intentional exposure of human subjects to a pesticide. The study is being conducted with 

the intention of submitting the resulting data to EPA under the Federal Insecticide 

Fungicide and Rodenticide Act (FIFRA). 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, and 

since the study will be conducted in California, the provisions of the California Code of 

Regulations, Title 3, §6710 would apply. The protocol will be reviewed by an 

Institutional Review Board (IRB).” (V2:7) 

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. See Attachment 5. 

(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. See Attachment 4. 

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

subjects? 

Literacy in English or Spanish is a criterion for inclusion. (V2:30) 

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

investigators and subjects? 

Two field research associates (Victoria Standart and Noé Galván) are fluent in both 

English and Spanish. (V2:9) 
Attachment 2 

Page 46 of 52 

All written materials provided to subjects will be available in both English and Spanish. 

(V2:14,20) A copy of the poster entitled “Controlling Heat Stress Made Simple” in 

English and Spanish will be posted in the dressing area at each site. (V2:20) 

“Therefore to adequately capture the ethnic diversity in the Fresno area, recruitment 

materials and all communications with potential subjects will be available in English and 

Spanish, as preferred by the subject.” (V2:27) 

“Janitorial services located in Fresno County and providing professional cleaning 

services for commercial buildings in the Fresno County, CA area will be contacted and 

asked to post flyers soliciting study subjects independently from the janitorial service. . . . 

The initial contact with service providers will determine language preference (English 

and/or Spanish) for the flyers.” (V2:27)

“If the firm wishes to post the flyer, research personnel will provide a general description 

of the study...and determine whether the flyer seems intelligible for the firm’s 

employees.” (V2:27) 

“The recruiting flyers will include telephone numbers for both English and Spanish 

speakers, and voicemail in the appropriate language will be available for messages when 

direct human contact is not possible.” (V2:28) 

“The recruiting advertisements [placed in area newspapers] will include a brief 

description of the study and include telephone numbers for English and Spanish speakers 

to call for more information.” (V2:28) 

“A Spanish-speaking member of the research team will be present during monitoring 

events involving subjects whose preferred language is Spanish.” (V2:32) 

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

discomforts? 

All written recruitment, consent, and risk communication materials will be available in 

both English and Spanish (including label, MSDS, recruiting materials, flyers, and poster 

entitled “Controlling Heat Stress Made Simple”). (V2:14, 20) 

“During the scheduled informed consent, the Principal Investigator or designee will share 

information on the study design with interested participants, and provide them with 

copies of the IRB approved Informed Consent Form (Appendix B) and answer their 

questions. The Principal Investigator or designee will describe the study to the individual 

in great detail and encourage each potential subject to ask questions and request 

clarification at any time during this process as well as in all activities that follow. The 

Principal Investigator or designee will provide each potential subject a copy of the 

product label (Appendix A), make available the MSDS (Appendix E) and answer any 

questions regarding the product to be tested. The Principal Investigator or designee will 

go over the Inclusion and Exclusion Criteria (see 9.A.iii. below) for the study and answer 
Attachment 2 

Page 47 of 52 

any questions that the potential subjects have. Potential subjects will be provided with 

copies of the Subject Self-Reporting Demographic Form (Appendix D) and the State of 

California Department of Pesticide Regulation “Experimental Subject’s Bill of Rights” 

(Appendix C) and asked if they would like to take any of the materials home to discuss 

with family and friends before deciding whether to enroll in the study. If the potential 

subject wishes to continue with the enrollment process at that time, the Principal 

Investigator or designee will explain to potential subjects they may withdraw from the 

research study at any time without penalty to their compensation. The Principal 

Investigator or designee will then read the “Experimental Subject’s Bill of Rights” to the 

potential subject. The amount and form of compensation, the potential risks and 

discomforts and treatment and compensation for injury will be more fully explained and 

potential subjects will be encouraged to ask questions. If the potential subjects do not 

have any questions and are interested in participating in this research study, they will then 

be asked to sign the Informed Consent Form and then fill out the Subject Self-Reporting 

Demographic Form.” (V2: 28-29) 

“A Spanish-speaking member of the research team will be available at recruitment 

meetings to assist and ensure communication with anyone preferring Spanish over 

English.” (V2:30) 

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

and obtain their consent? 

See procedure quoted in 8(f) above. 

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

coercion or undue influence? 

“Janitorial services providing professional cleaning services for commercial buildings in 

Fresno County, CA will be contacted by qualified research personnel and asked whether 

they would be willing to post a flyer soliciting volunteers for a study to be conducted 

independent of the janitorial service...If the firm wishes to post the flyer, research 

personnel will provide a general description of the study, explain the need and 

importance of remaining neutral (un-coercive) in their interactions with employees 

regarding study participation, and determine whether the flyer seems intelligible for the 

firm’s employees. Assuming the owner/manager still wishes to post the flyer, research 

personnel will provide approval to do so....To avoid the potential for coercion, subjects 

will not be recruited directly through contract janitorial service companies. Flyers will 

direct interested workers to contact research personnel directly.” (V2:27-28) 

9. Respect for Subjects 

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

their privacy? 
Attachment 2 

Page 48 of 52 

“Individual data, excluding the subject’s name and address, will be entered in Golden 

Pacific Laboratories’ computer data base by SISN. All subjects’ names and personal 

identifiers provided will be kept confidential to ensure their privacy.” (V2:31) 

“If a subject is female, she will be taken to a private area and asked to take a urine 

pregnancy test using an over-the-counter pregnancy test kit...Results of the pregnancy 

test will be kept in confidence, they will not be recorded, and they will be discussed only 

with the subject that provided the urine sample.” (V2:34-35) 

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

any time without penalty? 

See passage quoted in 8(f) above, and these passages from the consent document:

“If you decide to participate in this study it will be because you want to. There will be no 

direct benefit to you if you do decide to participate, and no harm to you if you decide not 

to. The choice is up to you.” (V2:72) 

“You are free to withdraw from this study at any time, for any reason. Simply tell any 

member of the research team. If you decide not to participate in this study or to withdraw 

from it, you will not be penalized in any way or lose any benefits.” (V2:73) 

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

dealt with? 

“All individuals that show up for the informed consent interview will be compensated 

$20 in cash at completion of the interview for their time and inconvenience. All 

individuals who are qualified, sign the informed consent form, and report to their 

assigned study site, will receive $100 in cash for their time and inconvenience when they 

leave the study site, whether they are monitored or not...Compensation will be provided 

to individuals who complete their assigned participation or who need to withdraw for 

whatever reason.” (V2:32) 

Subjects who are withdrawn by the investigators—and all participating subjects in the 

case that the entire study is stopped—are promised payment in full. (V2:73) 
Attachment 3 

Page 49 of 52 

§ 26.1111 Criteria for IRB approval of research 

AEATF II Aerosol Scenario/Protocol AEA04/070270: August 4, 2009 

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. 

Y 
Attachment 4 

Page 50 of 52 

§26.1116 General requirements for informed consent 

AEATF II Aerosol Scenario/Protocol AEA04/070270: August 4, 2009 

Criterion Y/N Comments 

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 

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 

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 

(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 

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

subject 

Y 

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

reasonably be expected from the research 

Y 

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

if any, that might be advantageous to the subject 

n/a 

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

identifying the subject will be maintained 

Y 

(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 Although research doesn’t involve 

more than minimal risk, compensation and treatment of injuries are 

provided for 

(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 

(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 

(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 

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

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

Y 

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

research 

Y 

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

and procedures for orderly termination of participation by the subject 

Y 

(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 

(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 
Attachment 5 

Page 51 of 52 

§26.1117 Documentation of informed consent 

AEATF II Aerosol Scenario/Protocol AEA04/070270: August 4, 2009 

Criterion Y/N Comments 

(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 

(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 

(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 6 

Page 52 of 52 

40 CFR 26.1125 Prior submission of proposed human research for EPA review

AEATF II Aerosol Scenario/Protocol AEA04/070270: August 4, 2009 

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 

(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 

V1: 6-56; V2:3-156 

V3:20-221 

V2:157-203 

(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 V2:157-159; V3:310-317 

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

(4) Copies of all correspondence between the IRB and the investigators. Y V3:4-317 

(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 

V3:316-317 

V3:316-317 

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

§26.1108(a) and §26.1108(b). 

Y Separately submitted to EPA 

under confidentiality claim 

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 

(1) The potential risks to human subjects Y V2:14-17 

(2) The measures proposed to minimize risks to the human 

subjects; 

Y V2:14-17 

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

research, and to whom they would accrue 

Y Nature -V2:14-17 

No discussion of magnitude 

(4) Alternative means of obtaining information comparable to what 

would be collected through the proposed research; and 

§1125(a) 

Y V2:13 

a discussion of: 

(5) The balance of risks and benefits of the proposed research. Y V2:17 

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

agreements as originally provided to the IRB, and as approved by the IRB. 

Y Original V3:80-90 

Approved V2:64-85; 182-203 

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

advertisements proposed to be used. 

Y V2:27-32; 96-100; 101-107 

§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 V2:27-32 

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

sponsors. 

The following Information, to the 

Y V3:4-317 

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 V2:157-159