Page 1 of 52 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 Scien ce Support Branch (RASSB)

Antimicrobials Division

REF: Selim, S. (2009) Transmittal Letter, 40 CFR 26.1125 Checklist, and Primary Documentation: Aerosol Application Scenar io: 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 Surf ace 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 Unpublished protocol dated August 4, 2009, prepared by Golden Pacific Laboratories for the Antimicrobial Exposure Assessment Task Force II under Sponsor ID AEA04 and GPL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 St udy. 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 E xposure Monitoring Program. August 4, 2009 ( Volume 4 )Page 2 of 52 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 th e 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 appe nded to this review as Attachment 6. All elements of required documentation are provid ed in the submitted protocol package.

Volume 1 of the submitted package incl udes 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: Ra tionale 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 Eng lish (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 Englis h (p. 93) and Spanish (pp. 94-95)

• IRB-approved Flyer soliciti ng research subjects, in E nglish (p. 97) and Spanish (p. 98)

• IRB-approved Newspaper advertisement so liciting research subjects, in English (p. 99) and Spanish (p. 100)

• Employer contact script and subject invitati on to participate script, in English (pp. 103-104) and Spanish (pp. 105-106)

• Executive Summaries from ADBAC a nd 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.Page 3 of 52 • 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 incl udes documentation of communications with IIRB, Inc. concerning the aer osol protocol, including:

• Protocol version transmitted to IIRB (b earing 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 im perative 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-s ubmitted 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 antim icrobial products that are applied by a multitude of application techniques incl uding from an aerosol spray can. Aerosol spraying can be accomplished with diffe rent types of nozzles, volumes, and techniques. AEATF II defines “...the aerosol application scenario is defined as the hand-held pressurized aerosol-based app lication of a label-specified end-use formulation containing an antim icrobial chemical.” (V1:8) 4 The actual task of spraying will be to spray a smooth, sweep ing, overlapping pattern until the treated surface is wet. The scope of the choices of aerosol can characteristics that are reasonably expected to influence exposur e includes nozzle size, particle size generated, ejection rate, and ge neration of non-volatile active ingredient. From this array of choices, the industry and regulato ry consensus was to monitor the selected product – Clorox Commercial Solutions Cl orox Disinfecting Spray (EPA Reg. No. 67619-03). A more detailed description of th e 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 numbe r. 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.”Page 4 of 52 provided below in Attachment 1 and Appe ndix 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 occupa tional 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 fo aming 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 ingredie nts. AD plans to use the data generated from the proposed aerosol study generica lly 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 o ccur in many Use Categories, most aerosol spray applications are expected to occu r in Use Categories II, III, IV, and V.

I Agricultural premises and equipment II Food handling/storage establis hments/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 scenar io is well defined, and we expect that resulting data will meet the needs of EPA and other regulatory agencies. The diversity of daily expo sures under the aerosol scenario as defined in this proposal will adequately describe a typical occupati onal 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 incorpor ated random elements where feasible. The AEATF II proposes to monitor dermal and inhalation exposures using passive dosimetry techniques to measure exposur e 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.Page 5 of 52 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.). Additionall y, the spray solution will be left on the surface, so no wiping will occur.

The amount of product applied during each m onitoring 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 w ill consist of the time it takes to spray a given number of cans. The focus on professi onal 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 ch ance that samples w ould be at or below the limit of detection of the analytical me thods used. Because of longer task duration and consequent exposure to greater quanti ties 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 re gistered aerosol spray uses include air sanitizers and foaming aerosols (e.g., bath tub and toilet cl eaners). The surface disinfecting spray was selected to represent the high end of potential exposure, based on a pilot study reported in the protocol s ubmission (V1:34). The AEATF II, in consultation with EPA/CDPR/PMRA, considered the various c onfigurations of aerosol spraying and the results of the pilot studie s’ nozzle size, particle si ze generated, and amount of material dispensed per unit time, air concen tration, and ejection ra te, and agreed that the hard-surface disinfection spray applica tion representing the reasonable high end of exposure. Other key factors considered in the selection of an aerosol application included its common use among consumers; it s use in tight spaces such as shower stalls; and the high exposure pot ential 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 su rface. Secondly, the AEATF II is developing other data under the “wipe” s cenario 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 pr oducts 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 establ ished. Subjects will apply the amount ofPage 6 of 52 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 act ual 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 se lected to be Fresno County, CA. EPA believes that the application of an anti microbial 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 laborato ry conducting the study, it is large enough to have a substantial population of professi onal 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 clus ters 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 th at each selected building must include appropriate building/room conf igurations, specifically hotels/motels with 20 or more units containing various full kitchens, kitche nettes, 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/mo tels 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 Yello wPages.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 protoc ol stipulating that if motels

with kitchens are not ava ilable 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 treatme nt 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.Page 7 of 52 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 kitchene tte and 20 or more units;

• Hotel/motel without kitchen/kitc henette 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. Theref ore, 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 subject s withdraw or fail to meet all eligibility criteria. Subjects will be professional janitors, selected from among those who respond to flyers posted in randomly select ed janitorial service companies in the Fresno area, and who meet the eligibility criteria for participation. In addition to the flyers, advertisements will be run simulta neously 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 di ffering 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 wi dely used, readily available, and there is a reliable and sensitive analyt ical 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 disc ussed in the Governing Document. The

AEATF II reviewed two data sets in determ ining 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-mo tion tasks. The existing data set for aerosol exposure data in PH ED indicates less variation th en 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 bette r 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.Page 8 of 52 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 Di sinfecting Spray (EPA Reg. No. 67619-03) in 19 ounce ready-to-use pressurized spray can s, 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 hab its of the test subj ect. 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 overlappi ng 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 Disi nfecting Spray which contains the active ingredients didecyl dimethyl ammonium chlo ride (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 produc t 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 exposur es 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 i ndoor surfaces. The sele ction 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 spra ying of indoor surfaces. Dermal and inhalation exposure will be measured usi ng whole-body dosimeters (inner and outer), hand and face washes, and personal air monitors . The Agencies are most interested in

the inner dosimeters to asse ss potential exposure. The out er dosimeters will add to the existing data base on the development of protection factors for single layer of clothing. The potential for foot exposur e 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 t ubes 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.Page 9 of 52 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 r ecovery analyses, travel recovery analyses, storage stability studies, and break-through analyses of the air samplers. Method

validation results should be incl uded in the final study report.

Primary components of the field recovery an alyses include: two fo rtification levels per matrix at the LOQ, triplicate samples pe r 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 st orage 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 da ta will be imported into the AEATF II database (BHED®) where they will be avai lable to regulatory agencies for later statistical analyses. The study documentati on will report a confid ence-interval-based approach to determine the relative accur acy 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 a ddresses 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 sample s for laboratory, field, and storage stability samplesPage 10 of 52 Additionally, the AEATF II has addressed the technical aspects provided in the applicable exposure monitoring guidelines (i .e. Series 875 Group A a nd OECD Applicator Guidelines) as well as Good Laboratory Practices (GLPs).

The following element in the protocol requires revision before the re search 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 Pr ograms, Antimicrobial Division), the

AEATF II committed to address deviati ons from the benchmark accuracy goal (i.e., k=3) as follows: “[I]f large de viations from the benchmark goals are observed in the aerosol study, AEATF II w ill, 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 pre ssurized 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 m eeting contemporary standards of quality and reliability.

2. Subject Selection: Twenty-four adult subjects will be recruited among professional janitorial workers of Fresno County. Partic ipants will self-iden tify in response to flyers posted in the workplaces of janitorial services companies and advertisements in three Fresno newspapers targeti ng different demographic groups.

The use of newspaper advertisements is a new recruiting method adopted by the AEATF II because workplace flyers were in effective during recruiting for the mop and wipe studies. Although more than 50 flyers soliciting subj ects for the mop and wipe studies were distributed to janitori al firms in Fresno County, no one attended either janitorial firm manager meeting he ld to explain the study. Many janitorial firms that reviewed the flyer declined to pos t, and there were very few responses from employees of those employers who did agree to post a flyer. In June 2009, thePage 11 of 52 AEATF II amended the mop and wipe protocols to allow for the use of newspaper advertisements in addition to workplace flye rs for recruiting. Since the amendment, subjects have been recruite d for the mop and wipe studi es through both methods. The AEATF II intends to employ both recruiting methods simultaneously for the aerosol study.

Callers responding to either the workplace fl yers or the newspaper advertisements will be screened, scheduled for informed consent meetings, and enrolled at the

volunteer’s convenience. A lthough 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 c oncluded that it is necessary to enroll subjects as they are processed. The AEAT F 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 dela ys 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 Fr esno 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. Empl oyees or relatives of employees of the investigators and of cleaning product manufacturers are also excluded from participation.

No potential subjects are from a vulnera ble population. Recruitment materials and interactions with potential subjects w ill be conducted in English or Spanish, depending on subject preference. To mi nimize 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, a nd will be used in fu ll compliance with the approved label. Risks to subjects include risk s 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 wear ing 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 testi ng. All identified risks are characterized as of low probability.

Risks are minimized by exclusion of candida tes 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 e nvironment; 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 proc edures to keep results of pregnancy testing private and to permit discrete w ithdrawal; provision of appropriate work clothing and PPE.

4. Benefits: This research offers no direct bene fits to the subjects, but subjects may request their individual resu lts, 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 derm al 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. Th e low residual risk is reasonable, in light of the likely benefits to society from ne w data supporting more accurate applicator exposure assessments for antimicrobial pr oducts applied with pressurized aerosol

cans.

6. Independent Ethics Review: The proposed research has been reviewed and approved by the Independent Investigationa l 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 prefer red 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 prefer ring 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 w ill refer to individual subjects only by an arbitrary code. Provision is made for disc rete handling of pregna ncy testing, required of female subjects on the day of testing. Candidates and subjects will be repeatedly reminded that they are free to decline to par ticipate or to withdraw at any time for any reason, without penalty.Page 13 of 52 F. Compliance with Applicable Ethical Standards

This is a protocol for third-party resear ch involving intentio nal 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 standa rds 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 Regulati ons 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 ad dressed before the research

is initiated:

• In the statement about compensation for res earch-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 insuran ce of a third party under which you are

covered.”

40 CFR 26 Subpart L, at §26.1703, as amende d effective August 22, 2006, provides in pertinent part:

EPA shall not rely on data from any rese arch involving intent ional exposure of any human subject who is a pregnant wo man (and therefore her fetus), a nursing woman, or a child.

The protocol requires that subj ects be at least 18 years old and excludes female subjects who are pregnant or lactating. Thus §26.1703 would notforbid EPA to rely on a study executed according to this protocol.

If the deficiency noted above is addresse d and the amended protocol is approved by the overseeing IRB, this research should meet the ethical standa rds 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 Pr otocol 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 ResearchAttachment 1

Page 15 of 52 EPA Scenario Review: AEATF-II Aero sol Application Scenario/Protocol

Title: AEROSOL APPLICATION SCEN ARIO: 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 scen ario to assess exposure as accurately as possible, while ensuring that any error does not underestimate exposure. Many varied antimicrobial products can be a pplied 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 surf ace disinfecting sprays, air sanitizers, and foaming aerosols. From this array of c hoices, the consensus among the Task Force and the three regulatory agencies was to select the pressurized disinfecting aerosol spray for

hard surfaces. The hard su rface “pressurized disinfecting ae rosol spray was selected to represent this group of spray products because it is a commonly used product type, the application process (spray no zzle actuation) is similar acr oss 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 uni t time, air concentrations, and aerosol characteristics, across products in the f our major aerosol categories (hard surface disinfecting sprays, hard surf ace foaming aerosols, soft surface sprays, air fresheners).

“The aerosol application scenario involves ap plication 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 ade quate amount for cleaning. Hard surfaceAttachment 1

Page 16 of 52 applications are typically sprayed until visibly “thoroughl y” wet per label direction.” “Surface applications are typically made in smooth, sweeping, overlapping patterns. Examples of ‘representative’ spra y application techniques that th is scenario is expected to capture include horizontal spraying movi ng upward and downward from the starting point to hard surfaces such as lamina te, 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 st udy. This separation of tasks will allow regulatory agencies to assess the exposure asso ciated with either pr ofessional “leave-on” products or other products requiring post-spray wiping.

The AEATF II study proposes to recruit only pr ofessional janitors. Janitors have been selected to provide longer task durations th en are typical for cons umers, to reduce the likelihood of undetectable residues on sampling materials. Thus, the scenario design will generate conservative data when used to es timate consumer application exposures.

The amount of product applied during each mon itoring 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 pred etermined. 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 aer osol application methods. These studies

have limitations that reduce their value for an antimicrobial-oriented generic database. Both of these studies involved aerosol appli cation for indoor residen tial crack and crevice insecticide treatment which may not be repr esentative of antimicrobial use patterns. Although both studies monitored ap plication 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 ch emical resistant gloves, a practice that is not typical for aerosol application of antim icrobials. 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 applicati on include only 5 monitoring events with measurable exposures, and only hand exposur es were detectable for these MEs. The application duration, only a fr action of which involved actu al aerosol spraying, rangedAttachment 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 pre ssurized aerosol scenario to provide diversity in sampling include (1) application equipm ent; (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 severa l 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 pr ocess 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 variab les were identified as ha ving 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 pr ocess was conducted in three phases. The first phase consisted of conducting an intern al membership survey that identified all of the members’ aerosol produc ts and characterized them by some of the influentialAttachment 1

Page 18 of 52 exposure variables. The next phase categori zed 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 i ngredient; 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 rang e 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 si ze distribution data ranged from 16 -164 μ m depending on the product type and method of particle size determination. The hard surface fine spray products had particle size distribution 40-

157 μ m. The surface disinfectants which are al so used as air fresheners/sanitizers and air treatment products had part icle size range of 16-87 μ m, and most of them had the smallest nozzle size. The foaming spray pr oducts had particle si ze distribution of 24-

164 μ m” (V1:43).

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

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

surface with a vertical and overhead orientat ion 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 th e air concentration in the breathing zone and dermal deposition.” (V1:43)

The second phase of the selection process pl aced all of the surv eyed antimicrobial aerosol products into the follo wing 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 pr oduct selection process, one re presentative 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 representa tive aerosol product in the exposure study.

“The results of the pilot study clearly s how that based on nozzle size, amount of material dispensed per unit time, air concen trations, and aerosol characteristics, theAttachment 1

Page 19 of 52 hard surface disinf ectant product, i.e., Clorox Disinfecting Spray ( EPA Reg. No. 67619-03), represents the high-end exposure scen ario and the product most likely to produce measurable exposure and would theref ore serve as the surrogate for the study entitled, “Measurement of Potential De rmal and Inhalation Exposure During Application of a Liquid Antimicrobial Pest icide 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 mentione d 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 differe nt 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 av ailable units containing full kitchens

B. Hotels/motels with 20 or more av ailable 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., k itchen or ‘kitchenette’) conf igurations which might be expected to impact exposure pot ential 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 prev ious 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 interference s with non-subjects... It also allows the focus to be on aerosol application only as oppos ed to the broad range of ja nitorial activities a subjectAttachment 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 ingred ient 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 whic h 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 fr om JohnsonDiversey indicates that a single individual would clean 20 rooms/day, whic h is considered to be a reasonable upper bound value. Therefore, the maximum am ount 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 mon itor 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 di versity. “Each selected worker provides his/her unique set of behaviors to the aer osol application ta sk...this study permits only one monitoring event per worker in or der to capture a larger diversity of application behaviors” (V1:17). The use of professional applicator s is believed to be a reasonable surrogate for the range of potential users that may apply aerosol antimicrobial products, including consumer s, school janitors, hospital janitors, commercial lodging janitors, employees of janitorial services firms, etc.

(b) How have random elements been incorpor ated into the scenar io sampling design?

“Surrogate worker selection: This proce ss results in a simple random sample of qualifying subjects from the volun teer pool. Note, however, thatthis is not technically the same as a random sample from the exis ting population of prof essional janitorial workers. By definition, volunteers are self-sel ected and could, in theory, have different

characteristics than non-volunteers. Such fine distinctions have littl e relevance in this case, however, because this is not an obser vational study of existi ng applicator-days. Because workers are randomly assigned to synthetic application-day conditions, the resulting MEs are still consid ered synthetic applicator-day s. Thus, any type of random sampling of just one ME component (e.g., applic ator in this case) pr ovides 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 ra ndomized. Next, these properties will be investigated in (random) order until N C 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 subj ects from the volunteer pool.” (V1:17)

“...workers are randomly assigned to synthe tic application-day conditions...” (V1:17) “The N=18 randomly sampled professional app licators will be randomly assigned to the N=18 combinations of building/room confi guration category and application volume. This provides a diverse set of MEs with respec t to three meta-parameters: (1) applicator, (2) types of rooms, and (3) product volume applied. ” (V1:31)

(c) What feasible opportunities to incorp orate 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 labe l-specified end-use formulati on containing an antimicrobial

chemical. This includes the task of actual aer osol 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 a pplications 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 hori zontal 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 hor izontal 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, refrigerat or, and food preparation sink (V1:30).

“The Clorox Disinfecting Spray will be appl ied by janitorial profe ssionals according to typical practices, i.e., spraying surfaces from a distance of approximately 6-10 inches in a manner to apply enough formulation to provi de an adequate amount for cleaning. Hard surface applications are typically sprayed until visibly ‘thoroughly’ wet per label direction. No wiping will be conducted as pa rt of this application scenario. Surface applications are typically ma de in smooth, sweeping, overlapping patterns. Examples of ‘representative’ spray applica tion methods that this study is expected to capture (i.e.,Attachment 1

Page 22 of 52 “horizontal spraying moving upward and downw ard from the starting point to hard surfaces such as laminate, tile, porcelain, glass, and metal”) are specif ied 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 sp ray 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 specia lized 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 practi ce, aerosol application may or may not involve follow-on tasks, such as wiping th e sprayed surface. Some aerosol products are considered ‘leave-on’ sprays that do not re quire post-spray wiping. Only the actual aerosol spray application is c overed by this scenari o. Applicator exposur e associated with wiping is being addressed in another AEATF II scenario. Ther efore, 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 th e 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 Sp ray, is an appropriate choice for the development of surrogate exposure data becaus e it is stable, is characterized by a low vapor pressure (estimated to be 2E-11 mmHg), diluted solutions are not dermally irritating, no systemic toxici ty was seen in a 90-day de rmal rat study, the product is registered for aerosol sprays, and its prin cipal 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 (referre d to as Clorox Disinfecting Spray in this protocol), containing didecyl dimethyl am monium chloride (DDAC), n-Alkyl (C12, C14, and C16) dimethyl benzyl ammonium ch lorides (ADBAC), octyl decyl dimethyl ammonium chloride (ODAC), and dioctyl dimethyl ammoni um chloride (DODAC). The quaternary ammonium antimicrobials are co mmonly 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. Cl orox 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, a nd DDAC have complete toxicology databases with low mammalian toxicity. Virt ually all quat antimicrobial products contain more than a single quat, i.e., a readily av ailable product containi ng only ADBAC was not apparent. The analytical me thod 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 matric es to be used in this study is ongoing (GPL, 2009; in progress).” (V2:19)

“The very sensitive and selective analytic al method developed for the analysis of ADBAC on different study matrices will allo w 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 generati ng exposure data for other an timicrobial pesticides.” (V2:20)

ADBAC and DDAC have been approved fo r use in many formulations, and are extensively used in many janitorial produc ts. 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 establishe d through long term professional use of the product. The product will be us ed according to its label.

4. What is the rationale for the prop osed 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 a ppropriate 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 combin ation 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 areAttachment 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 th is scenario, the AEATF II pref ers to assume GSD=2.9 for the purposes of determining sample size. As al so 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) fo r a review of the results of the accuracy given n=2 versus n=6 MEs per cluster.

“Benchmark objectives specify accuracy goa ls that must be achieved within the framework of the reference sampling model wh en sample size is adequate. In this study, ‘sample size’ means both the number of cluste rs (NC) and the number of MEs per cluster (NM). For the aerosol a pplication 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 Aero sol Application Scenario/Protocol

Title: A Study For Measurement of Potential Dermal and Inhalation Exposure During Application of a Liquid Antimi crobial 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 applic ation monitoring study is to develop more accurate information on worker exposures to antimicrobials . . . that broadly represent those expected for the future application of arbitrary antimi crobial pesticides.” (V1:13)

“The main goal of the study is to genera te 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 [a ntimicrobial products in] aerosol cans.” (V1:38)Attachment 2

Page 26 of 52 (b) What research question does it addre ss? Why is this question important?

Would the research fill an im portant 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 professi onal 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 bei ng used by regulatory agencies

in the case of antimicrobials is the Chemical Manufacturers A ssociation 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 lim its the conclusions that can be made from the CMA study. Therefore, the Agency is requir ing 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 we re also echoed by regulatory agencies in California and Canada. All note that the exposur e data cannot be used as generic data for

all antimicrobials because recoveries were lo w, precision of the measurements were not established, and CMA did not establish the validity of generalizing the information among applications and end-us e 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 ha nd-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 b ecause they will normally be exposed to the test material when performing their daily ac tivities. There are no acceptable methods or

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

Page 27 of 52 2. Study Design

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

is it?

“The primary purpose of the aerosol applic ation 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 expect ed for the future appl ication of arbitrary antimicrobial pesticides.

Although this study will only monitor a sing le active ingredient, AEATF II and regulatory agencies generally recognize tw o important principles that allow such exposure results to be generalized to a larger set of conditions:

1. Dermal and inhalation exposure to antimic robial 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 predic t exposure for other active ingredients.

2. The principle of proportionality of exposure to appropriate measures of active ingredient contact po tential. 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 result ing database will contain sufficient data to support exposure assessments for aerosol applic ation for a number of antimicrobial active ingredients over a range of AaiH levels.” (V1:13)

“Benchmark objectives specify accuracy goa ls that must be achieved within the framework of the reference sampling model wh en sample size is adequate. In this study, ‘sample size’ means both the number of cluste rs (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 arithm etic mean and 95th pe rcentile 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

Page 28 of 52 2.1 Statistical Design

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

The goal is to use these data to characteri ze some ‘population’ aspect of the future exposure to arbitrary antimicr obial 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 stud y will utilize a combination of random sampling, randomization, and diversity selection methods. While this methodology contains some elements of all three pure situa tions ....[statistical theory], none apply completely. The ultimate goal of this study is to construct synthetic MEs that can be

used to characterize the dive rsity of future daily exposur es 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 se lection actually used. This reference model assumes

that:

1. Exposure, normalized by the amount of active ingredient handled, is lognormally distributed with a known ge ometric standard deviation (GSD). Equivalently, the logarith m 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 95 th percentile of normalized exposure are

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

“A Monte Carlo simulation approach was us ed to examine the impact of number of clusters (NC) and number of MEs per cl uster (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 clus ter 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 bett er indicator of the expected true relative variation forAttachment 2

Page 29 of 52 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 propos ed. 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 t reatment 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 spraye d will be executed first; a subject who is unable to continue spraying for the give n 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 ta rget 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 statis tical analysis of the data?

“The AEATF II will not statistically anal yze 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 turnove r rate, etc.) However, regulators and

other users of the constructed databa se (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 in clusion in BHED scenario monographs. Two specific types of analyses will be performed (these analyses are discussed in more

detail in the AEATF II’s Govern ing Document (AEATF II, 2008).Attachment 2

Page 30 of 52 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 exposur e normalized by amount of ai handled.

2. Cluster effects. The intraclass correla tion for clusters (ICC) and its confidence interval will be estimated using a vari ance components model. In addition, the effects, if any, of ignoring clusters in the estimation of m eans and percentiles will be determined by comparing the estim ates of a no-cluster model to those

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

(g) Are proposed statisti cal methods appropriate to an swer 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 jan itors elsewhere, and th at the exposures of professional janitors sprayi ng 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 aer osol 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 contai ns 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 duri ng 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 app lication scenario. Surf ace applications are typically made in smooth, sweeping, overlapping patterns. Examples of “representative” spray application techniques that this scenario is expected to capture include hori zontal spraying movingAttachment 2

Page 31 of 52 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 ba throoms and/or food preparation areas which

include horizontal and vertical surfaces (e.g. shower stall/tu b, 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 count er tops and some appliance surfaces. All

interior surfaces of the s hower/tub, the horizontal surface of the countertop/sink, and exterior surface only of toilets will be spraye d with the test substance. One complete can of aerosol product will cover the shower, sink and toilet in approxi mately 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 typica l 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 choi ce of test material and formulation?

“Based on the nozzle size, amount of ma terial dispensed per unit time, air concentrations, and aerosol ch aracteristics, across products in the four major aerosol categories (hard surface disi nfecting 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 produ ce measurable exposur es for purposes of the aerosol applicati on study.” (V1: 9)

“C14 ADBAC is the active i ngredient selected for measurement, based on its stability, abundance in the formulation, a nd 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 wa s selected as the analyte based primarily upon its abundance in aerosol produc ts, 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 wa s not apparent.” (V2:19)

“The analytical method for ADBAC on the pr oposed monitoring matrices at very low concentrations has been validated (GPL , 2004). The freezer storage stability of

ADBAC on the different matric es to be used in this study is ongoing (GPL, 2009; in progress).” (V2:19)Attachment 2

Page 32 of 52 “The very sensitive and selective analytic al method developed for the analysis of ADBAC on different study matrices will allo w 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 cont ains several active ingredients. However, only one of these, C14 ADBAC, will be qua ntified. Because the concentration of

ADBAC in the aerosol canisters is the same for all MEs, the amount of active ingredient ‘handled’ is vari ed 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 gram s of formulated product). (V2:25)

ADBAC was selected because of its relative ly 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 durat ion to apply the specified amounts will range from 30 to 180 minutes. The tota l 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 handl ed 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 can isters are allowed to stop and as long as at least one canister is appl ied the dosimeters will be analyzed. If the subjects fail to spray a single canister, th e ME will not be used and the monitoring media will not be analyzed. (V2:26)

The AEATF II cites existing su rvey data the Antimicrobi al Exposure Joint Venture (AEJV) and information provided by J ohnsonDiversey which was based on was based in part on information from the fo llowing sources: 1) the American Hospital Association (http://www.a ha.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 aver age of ~113 g product per room (kitchen or bathroom) (or ~20% of a 19 oz can). In formation from JohnsonDiversey indicates that a single individual w ould clean 20 rooms/day, whic h 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 s ubstance will be measured externally using whole body inner and outer dosimeters, ha nd washes, and face/neck wipes. All monitored subjects will wear the outer dos imeter (representative outer clothing consisting of cotton long pants and cotton long sleeve shirts) dire ctly 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). Pote ntial exposure to resp irable, thoracic and inhalable particles (100, 10 and 2.5 μ m, respectively) will be characterized with a three stage RespiCon TM 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 performe d to determine if there are background ADBAC air concentrations. (V2:34)

“Airflow of pumps attached to the OV S tube and RespiConTM sampler will be calibrated to a nominal flow rate of appr oximately 2 and 3.1 L/min respectively usingAttachment 2

Page 34 of 52 SOPs of field study procedures. The begi nning flow rate of each pump will be checked and documented just before bei ng fitted to the subject.” (V2:35, 36)

“The inner and outer dosimeters, OV S tubes, filters from the RespiCon TM Particle Sampler, hand wash solutions, and face/neck wipes will be analyzed for residues of C14 ADBAC using validated an alytical methods.” (V2:12)

“ The amount of test substance applied during each ME will be determined by the total change in weight of a ll canisters used in the ME. E ach canister will be uniquely identified prior to experimental start. Pr ior 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 completi on of the ME, each canister used in the ME will be individually weighed, and the e nd 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 ne cessary to reconstruct the amount of test substance applied by each ME will be documented.” (V2:22)

“Air temperature and relative humidity of th e work area for the duration of exposure monitoring will be documented with au tomated 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 meas urements are accurate and reliable?

The AEATF II SOPs provide specific proce dures to ensure accurate measurements

such as calibration of inhalation mon itoring 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 Certif icate of Analysis will be kept in the raw data file.” (V2:19)

“Full details regarding fi eld recovery evaluation pr ocedures for all sampling media are given in the most recent vers ion of the SOPs of the AEATF II-8E[.1].” (V2:41)

(c) What QA methods are proposed?

Volume 2, Section 12, Analytical Procedur es, 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 con centrations 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 Pr ograms, Antimicrobial Division), the

AEATF II committed to address deviations from the benchmark accuracy goal (i.e., k=3) as follows: “[I]f large deviations fr om the benchmark goals are observed in the aerosol study, AEATF II will, in consultati on 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 estim ates into account. It is possible that collection of a dditional clusters might be c onsidered.” (a copy of the

letter is included in th e supplemental materials).

3. Subject Selection

3.1 Representativeness of Sample

(a) What is the population of co ncern? 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 th e janitorial/cleaning service population of Fresno County. The most-recent US Census i ndicates that 40% of the population in Fresno, CA metropolitan area is Hispanic. The proportion of Hispanics in service industries, e.g., janitorial se rvices, 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 poten tial 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 Ca lifornia Advocate and the Fresno edition of Vida en el Valle. The Fresno Bee is a larg e, general circulation daily paper in Fresno County. The California Advocate is the dominant African American communityAttachment 2

Page 36 of 52 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 representativ e of the population of concern? If not, why not?

Expected participants will self-identify in response to flyers so liciting interest, posted in the workplaces of janitorial services comp anies, or in response to advertisements placed in three Fresno-area newspapers. Vo lunteers may differ in unknowable ways from other workers who do not step forw ard. 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 th e Governing Document (2008), intends to develop a database of exposure monitoring data that can be used to support practical regulatory decisions about future exposu res for different (including currently nonexistent) active ingredient s and their associated produc ts. The database needs to address a variety of exposure scenarios for which no or limited data currently exist. The aerosol application scenar io is an important component of the AEATF II program and the focus of this study. As noted in th e previous section, existing monitoring data for this scenario are considered inadequate.

“The primary purpose of the aerosol applic ation monitoring study is to develop more accurate information on worker exposures to antimicrobials. These data will consist of dermal and inhalation exposure estima tes derived from monitoring subjects under conditions that broadly represent those expect ed for the future appl ication of arbitrary antimicrobial pesticides.

“Although this study will use only a si ngle active ingredient, AEATF II and regulatory agencies generally recognize tw o important principles that allow such exposure results to be generalized to a larger set of conditions:

1. Dermal and inhalation exposure to an timicrobial chemicals are considered generic (i.e., independent of the part icular 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 poten tial. For example, if measured exposure is E 1 when the amount of active ingredient handled (AaiH) is H 1 , then the predicted exposure

when AaiH is H 2 is just E 2 = H 2 (E 1 /H 1 ).Attachment 2

Page 37 of 52 “Consequently, AEATF II anticipates the re sulting database will contain sufficient data to support exposure assessments fo r 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-specifi c 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 c onditions. Therefore, the ae rosol application scenario can be viewed as the collection (or ‘populat ion’) 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 monito ring event, AEATF II researchers will collect dermal and inhalation exposure in formation from a worker while he/she performs aerosol application. Each ME is de signed 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 AE ATF II scenarios) only a small number of expensive experimentally-obtained monito ring 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 Consen t 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 Consen t Form or the Subject Self Reporting Demographic Form

• Does not read and understand English or SpanishAttachment 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 in farcts, stroke, congestive

heart failure or uncontroll ed high blood pressure)

• Is an employee of, or is related by blood or marriage to an employee of Golden Pacific Laboratories, Grayso n | Eurofins, or a cleaning product

manufacturer.”

(b) What, if any, is the relationship be tween 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 recruit ing 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 fl yer soliciting volunteers for a study to be conducted independent of the jani torial service...If the firm wishes to post the flyer, research personnel will provi de 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. Assu ming 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 info rmed consent form, and report to theirAttachment 2

Page 39 of 52 assigned study site, will rece ive $100 in cash for their time and inconvenience when they leave the study site, whet her 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, th e disqualified subject will be paid $100 as if a pa rticipant 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 ti me and incidental expenses incurred in study participation. Compensation will be pr ovided to individuals who complete their assigned participation or w ho 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, w ill receive $100 in cash...when they leave the study site, whether they are m onitored 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-register ed, 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 possi bly heat-related illness associated with wearing two layers of clothing; of discom fort 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

Page 40 of 52 Risks discussed in the consent form include ri sk of a reaction to th e test material, risk of discomfort, risk of sti nging from alcohol wash and wi pes, a small possibility of heat stress, risk of embarrassment, possible su rprise at the results of a pregnancy test,. (V2:70-71)

(c) What is the probability of each risk asso ciated 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; excl usion 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 cont rolled-temperature environment; alerting subjects to signs and symptoms of heat st ress; monitoring heat index with associated stopping rules; limited time of exposure with re st periods at 30-min intervals, or more frequently if requested; close observati on of subjects; training of experienced technicians to minimize embarrassment; incorp oration of procedures to keep results of pregnancy testing private and to pe rmit 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 Commerci al Solutions Clorox Disinf ecting Spray (EPA Reg. No. 67619-03) contains the test substance, ADB AC, and another activ e ingredient (i.e., DDAC). Therefore, the established NOAELs for both ADBAC and DDAC are provided below.

The ADBAC risk assessment developed to support the Reregi stration Eligibility Decision (RED) document provides for the se lection of the toxicological endpoints for risk assessment purposes. The dermal toxicological endpoints indicate no systemic toxicity. However, dermal irri tation 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 derm al endpoint selected from the 21-day study is 333 ug/cm 2 and 80 ug/cm 2 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 duratio ns is based on a developmental toxicityAttachment 2

Page 41 of 52 study in rabbits (MRID 42392801) with a NOA EL of 3 mg/kg/day. The inhalation risk assessment for aerosol inhalation expos ure 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 Reregist ration Eligibility Decision (RED) document provides for the se lection of the toxicological endpoints for risk assessment purposes. The dermal t oxicological endpoints indicate that low concentrations of DDAC (0.13% ai tested in a 21-day de rmal toxicity study, MRID 45656601) display no dermal irritation eff ects 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 concen tration of 0.378% activ e ingredient (i.e., aggregate of octyl dimethyl ammonium chloride, dioctyl dimethyl ammonium chloride, and didecyl dimethyl ammonium chlo ride) will not trigger a risk of concern. The inhalation toxicological endpoint iden tified for DDAC for all exposure durations is based on two oral toxicity studies (prena tal 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 inha lation 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 medi cal 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 provi de for safety monitoring?

“If a subject reports an adverse sk in reaction during the work period, they will be asked to immediately stop wo rking. 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 possibi lity of heat stress, the study will be conducted indoorsAttachment 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 s ubjects’ 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 resear ch personnel at the field site.

“In brief, researchers will observe subjects for possible signs of early heat illness such as fatigue, dizziness, ir ritability, or decreased concentration, especia lly 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 mome nt. 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 woul d 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 app ears 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-e xposure 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 Inves tigator 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 st udy during or following conduct of the study.” (V2:32)

“If two or more subject develop an advers e skin reaction after they leave the study site, all subjects will be cont acted by the Principal Investigator to determine whether further medical management is appropriate.” (V2:33)

(g) How and by whom will medical care for re search-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 ca re. If necessary, we will take you there. We will pay for need ed medical treatment that is not paid for by your own insurance or by someone else. To find out more, or if you think youAttachment 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 insuran ce 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 s ubjects 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 anticipate d 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 bacter ia and viruses known to produce increased morbidity and mortality in humans, domes tic animals and pets...Measuring exposure of

workers in this research st udy 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 predic t 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 benefi ts be distributed? Who woul d benefit from the proposed

research?

“Results from the study may benefit EPA...by reducing uncertainty about the range of exposure experienced by consumers and worker s 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 iden tified societal benefits would be realized?

The research is very likely to produce more accurate and reliable information concerning exposure in the aerosol scenari o, with resulting societal bene fits in the form of more

accurate and confident assessments of applicator exposure and risk.

6. Risk/Benefit Balance: How do the risk s to subjects weigh against the anticipated benefits of the research, to subjects or to society?

The likely benefit to janitorial workers as a whol e 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 participan ts. Aerosol antimicrobial products are widely used both by janitorial workers and the general public. Exposure data for this scenario meeting contemporary standards of re liability 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 st udy, 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 se rious adverse effects is very small. In summary, the risks to study participants from par ticipating in this study are reas onable 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 Federa l-Wide Assurance from OHRP?

No.Attachment 2

Page 45 of 52 (f) Are complete records of the IRB rev iew 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 thir d-party research involving what EPA has interpreted to be intentional exposure of human s ubjects to a pesticide. The study is being conducted with the intention of submitting the resulting da ta 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 a nd 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 obtain ed from each prospective subject?

Yes.

(b) Will informed consent be appropri ately documented, consistent with the requirements of 40 CFR §26.1117?

Yes. See Attachment 5.

(c) Do the informed consent material s meet the requirements of 40 CFR §26.1116, including adequate characterization of th e risks and discomforts to subjects from participation in the research, the potential be nefits 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 overco me language differences, if any, between investigators and subjects?

Two field research associates (Victoria St andart and Noé Galván) are fluent in both English and Spanish. (V2:9)Attachment 2

Page 46 of 52 All written materials provided to subjects w ill be available in both English and Spanish. (V2:14,20) A copy of the poster entitled “C ontrolling Heat Stress Made Simple” in English and Spanish will be posted in th e dressing area at each site. (V2:20)

“Therefore to adequately capture the ethni c diversity in the Fresno area, recruitment materials and all communications with potentia l 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 Fr esno County, CA area will be contacted and asked to post flyers soliciting st udy subjects independently from the janitorial service. . . .

The initial contact with serv ice providers will determine language preference (English and/or Spanish) for the flyers.” (V2:27)

“If the firm wishes to post the flyer, resear ch personnel will provide a general description of the study...and determine whether the fl yer 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 la nguage 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 in clude telephone numbers for E nglish 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 pref erred language is Spanish.” (V2:32)

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

discomforts?

All written recruitment, consent, and risk co mmunication materials will be available in both English and Spanish (including label, MSDS , recruiting materials, flyers, and poster entitled “Controlling Heat Stre ss Made Simple”). (V2:14, 20)

“During the scheduled informed consent, the Principal Investigator or designee will share information on the study design with interest ed participants, and provide them with copies of the IRB approved Informed Cons ent Form (Appendix B) and answer their questions. The Principal Investigator or desi gnee will describe the study to the individual in great detail and encourage each potent ial subject to ask questions and request clarification at any time duri ng this process as well as in all activities that follow. The Principal Investigator or designee will prov ide 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 (s ee 9.A.iii. below) for the study and answerAttachment 2

Page 47 of 52 any questions that the potential subjects have . Potential subjects will be provided with copies of the Subject Self-Reporting Demogr aphic Form (Appendix D) and the State of California Department of Pesticide Regulati on “Experimental Subject’s Bill of Rights” (Appendix C) and asked if they would like to take any of th e materials home to discuss with family and friends before deciding whet her to enroll in the study. If the potential subject wishes to continue with the enro llment process at that time, the Principal Investigator or designee will explain to poten tial 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 “E xperimental 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 as k questions. If the potential subjects do not have any questions and are intere sted 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 communi cation 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 ensu re 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 qualif ied research personnel and asked whether they would be willing to pos t 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 descript ion of the study, explain the need and importance of remaining neutral (un-coercive ) in their interactions with employees regarding study participation, and determine wh ether 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 serv ice companies. Flyers will

direct interested workers to contact re search 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 confidenti al 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-t he-counter pregnancy test k it...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 freed om to withdraw from the research at any time without penalty?

See passage quoted in 8(f) above, and th ese passages from the consent document:

“If you decide to participate in this study it wi ll be because you want to. There will be no direct benefit to you if you do decide to par ticipate, 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 a ny 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 tim e and inconvenience. All individuals who are qualified, sign the info rmed consent form, and report to their assigned study site, will receiv e $100 in cash for their time and inconvenience when they leave the study site, whether they are mon itored or not...Compensation will be provided to individuals who complete their assigned pa rticipation 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—ar e promised payment in full. (V2:73)Attachment 3

Page 49 of 52 § 26.1111 Criteria for IRB approval of research
AEATF II Aerosol Scenario/Proto col AEA04/070270: August 4, 2009

Criterion Y/N Comment/Page Reference
(a)(1)(i) Risks to subjects are minimized by us ing 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 re asonable in relation to antici pated benefits, if any, to subjects, and the importance of the knowle dge 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 resear ch (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 appl ying knowledge gained in the research (for example, the possible effects of the re search 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 in volving 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 prot ect the privacy of subjects and to maintain t he 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. YAttachment 4

Page 50 of 52 §26.1116 General requirements for informed consent AEATF II Aerosol Scenario/Proto col 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 t he legally effective informed consent of the subject or the subject’s le gally authorized representative Y

An investigator shall seek such consent only under circumstances that provide the prospective subject or the representative suffici ent 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 ap pears to release the investigator, the sponsor, the institution or its agents fr om 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 proced ures to be followed, and identification of any procedures which are experimental Y

(2) A description of any reasonably fo reseeable 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 alternativ e 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 involvemore 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 subjec ts’ rights, and whom to contact in the event of a research-related injury to the subject Y(a) In seeking informed consent the following informationshall be provided to each subject

(8) A statement that participation is vol untary, refusal to participate will involve no penalty or loss of benefits to which t he subject is otherwise entitled, and the subject may discontinue participation at any time without penalty or loss of benefits to which the subj ect is otherwise entitled Y

(1) A statement that the particular treatme nt or procedure may involve risks to the subject (or to the embryo or fetu s, if the subject may become pregnant) which are currently unforeseeable Y

(2) Anticipated circumstances under wh ich the subject’s participation may be terminated by the investigator with out regard to the subject’s consent Y

(3) Any additional costs to the subject t hat 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 ormore of the following elementsof information shall also beprovided 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. YAttachment 5

Page 51 of 52 §26.1117 Documentation of informed consent AEATF II Aerosol Scenario/Proto col 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 gi ven to the person signing the form. Y

(b)(1) The consent form may be a writt en 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 authoriz ed 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 r epresentative. 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 subjec t or the representative. Only the short form itself is to be signed by the subject or t he 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 co py of the summary shall be given to the subject or the representative, in add ition to a copy of the short form. n/aAttachment 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 hu man research covered by §26.1101(a) shall, after receiving approval from all appropriate IRBs, submit to EPA prior to initiating such research all information relevant to the proposed research specified by §26.1115(a), and the following addition al information, to the extent not already included:
Requirement Y/N Comments (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 discus sion 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, fu ll-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 claimAll 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 Y V2:13§1125(a)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 t he IRB, and as approved by the IRB. Y Original V3:80-90 Approved V2:64-85; 182-203 §1125(c): Information about how subjec ts 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. Y V3:4-317The following Information, to theextent 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