February 21, 2010 

Tim Leighton 

Office of Pesticide Programs 

U.S. Environmental Protection Agency 

One Potomac Yard (South Building) 

2777 South Crystal Drive 

Arlington, VA 22202 

Dear Tim: 

This Document summarizes changes to the aerosol study protocol following the October 20-21, 

2009 HSRB meeting and December 16, 2009 HSRB final report. A revised protocol with track 

changes is attached. The changes made to the protocol fall into four categories: 1) changes in the 

study personnel; 2) changes to the study design; 3) changes in ethics sections; and 4) justification 

for separating monitoring activities between clusters from seven days to two days. 

1. Changes in Personnel 

Grayson/Eurofin personnel were replaced with the Golden Pacific Laboratories (GPL) 

personnel throughout the protocol. 

2. Changes in Study Design Sections

In each of the following sections a) through d), HSRB comments are italicized followed 

by an explanation of how the AEATF has addressed them in a revised protocol. 

a) Sample Size and Analysis: Raw and descriptive data will be provided, but no 

statistical analyses have been planned. Sample size adequacy cannot be judged 

without a statistical analysis plan. Placing 18 subjects into three clusters of six each 

requires consideration of cluster effects, and may complicate analysis of the results. 

Assumed constants related to exposure to the active ingredient may also prove to be 

incorrect, and additional subjects may need to be enrolled. 

If the benchmark accuracy goal (i.e., k=3) is not met once the data are collected and 

analyzed, the AEATF II will, in consultation with regulatory agencies, determine the 

best course of action to take. This may mean the development of guidance for the use 

of these data that takes the increased imprecision of the estimates into account. It is 

possible that collection of additional clusters might be considered. See Data Analysis 

Section 12 E page 53. 
Tim Leighton 

Summary of changes to the Aerosol Study Protocol 

February 21, 2010 

Page 2 

b) Application of Spray: The target number of cans sprayed in each location by each 

worker on each day could be specified better within the protocol. For example, does 

a worker need to continue spraying a room if the target application is met before 

completion of the room? Additional protocol clarifications needed include: 

addressing the issue of the interval between sampling at different sites (currently, it 

appears that no two monitoring events can occur within the same building within one 

week); clarifying the issue of using empty apartments rather than motel rooms; and 

clarifying the effect of exhaust vent effects on exposure, which could be different than 

indicated in the protocol. 

No more than 2 MEs per day could be monitored at a given site on a given day, and 

will depend upon the number of rooms available as well as on the number of canisters 

assigned. Rooms at a given site will only be used once per day. Subjects in the A, C, 

and F groups will be provided with half full canisters to easily fulfill the assigned 

amount without going over it. See Study Design Section 8B, page 26. 

c) Setting the minimal spike to two to four times that of the limit of quantitation (LOQ; 

cf. AEATF II 2009b: 42, 46). 

This is covered in two sections. A table summarizing the LOQ (see below) was 

added Under Test Substance Section 7B, page 21. 

Matrix LOQ 

Air Sampling Tubes 25 ng/tube 

Hand Washes 1.0 ng/mL 

Face/Neck Wipes 100 ng/sample 

Inner Dosimeter Section 3.0 μg/sample 

Outer Dosimeter Section 3.0 μg/sample 

Respicon Fiberglass Filters 25 ng/filter 

A second table (shown below) showing the 4X fortification levels was added under 

Field Recovery Evaluation Section 10H page 47. 

Matrix Fortification Level 

Air Sampling Tubes 100 ng/sample and 2.0 μg/sample 

Hand Washes 4.0 and 400 ng/mL 

Face/Neck Wipes 400 ng/sample and 10 μg/sample 

Inner Dosimeter Section 12 μg/sample and 1.0 mg/sample 

Outer Dosimeter Section 12 μg/sample and 1.0 mg/sample 

Respicon Fiberglass Filters 100 ng/filter and 2.0 μg/filter 
Tim Leighton 

Summary of changes to the Aerosol Study Protocol 

February 21, 2010 

Page 3 

d) Considering other variables that may influence the measurement of exposure, 

including whether the surface being treated is dry or wet at the time of the next spray 

application, and whether the applicant accidentally wiped the surface after spraying. 

Data accumulated during gross deviations from the protocol should also be excluded. 

Finally, the protocol should indicate that there will be a maximum of two workers on 

any given day in the same location. 

See Random Selection of Facilities as Monitoring. Section 8 B, page 26. 

No more than 2 MEs per day could be monitored at a given site on a given day, and 

will depend upon the number of rooms available as well as on the number of canisters 

assigned. Rooms at a given site will only be used once per day. 

See Assignment of Amount of Active Ingredient Handled to MEs. Section 8D, page 

27. 

The total number of qualified subjects will each be assigned a consecutive number, 

starting at AE1. The numbers will then be randomized using a research randomizer 

program accessible at the following internet website: http://randomizer.org. The first 

24 numbers in the generated randomized list will determine the participating subjects, 

while the remaining subjects will be held as alternates, their order for potential entry 

into the study being determined by the randomization process. The 24 subjects will 

be split into three groups, each corresponding to the location of one of the three 

experimental sites. The first set of eight numbers (AE1-AE8, where AE indicates the 

randomized order list) will be grouped into location A, the second set of eight 

numbers (AE9-AE16) will be grouped into location B, and the third set of eight 

numbers (AE17-AE24) will be grouped into location C. There will be six different 

aerosol spraying durations for assigned monitoring events (MEs). Within each group 

of eight, the first six subjects will be assigned the durations listed under section 8.C in 

the protocol (AE1 1 to 1.5 canisters; AE2 1.5 to 2 canisters; AE3 2 to 2.5 canisters; 

AE4 2.5 to 3 canisters; AE5 3 to 3.5 canisters; AE6 3.5 to 4 canisters). The last two 

subjects in the group of eight will be considered as alternates and will be on hand if 

any subject does not show up. If additional subjects above the 24 initially selected 

are required, AE25 will be contacted followed by AE 26 and so on, until a total of 18 

MEs are completed for the study. 

Once the subjects have been randomized into three clusters, subjects from the first 

cluster will be scheduled into the study. Every attempt will be made to schedule two 

subjects per day. No more than two subjects will be monitored in one day. An 

example schedule could look as follows: 

E

v

e

r 

Day ME No. of Canisters ME No. of Canisters 

1 AE6 3.5 to 4 AE1 1 to 1.5 

2 AE5 3 to 3.5 AE2 1.5 to 2 

3 AE4 2.5 to 3 AE3 2 to 2.5 

4 AE7 alternate AE8 Alternate 
Tim Leighton 

Summary of changes to the Aerosol Study Protocol 

February 21, 2010 

Page 4 

Every attempt will be made to schedule the subjects with the longest duration first. If 

this is not possible because of work or scheduling conflict, subjects with the longest 

MEs will be assigned to the earliest time slot available. If the subject with the longest 

ME cannot finish his or her application volume, subsequent subject with a shorter 

unfilled application volume will be asked to complete the longest ME. As long as the 

six subjects achieve the target spraying amount, the process is continued and six ME 

have been obtained one for each of the six strata. If this is not achieved, because two 

subjects ended up with the same spray amount, alternate subjects will be asked to 

participate in the study and fill the missing ME. 

3. Changes in the Ethics Section 

a) Possible subjects for this study may be undocumented immigrants. Recruitment 

materials should state that government-issued identification is necessary for 

enrollment in this study. 

The Principal Investigator or designee will check the potential subject’s driver license 

or government-issued identification card to verify identity as required by California 

DPR. See Subject Recruitment, Section 9 ii, page 32. 

b) A valid government-issued form of identification is necessary for enrollment. 

Volunteers who lack proper identification are excluded from study participation but 

no other action is taken, minimizing any social risks to undocumented workers from 

disclosure of their status to law-enforcement or immigration authorities. 

Volunteers who lack proper identification will not be enrolled in the study. No other 

action will be taken. Language added to the protocol. See Subject Recruitment, 

Section 9 ii, page 32. 

c) The Board agrees with the EPA ethics review that the informed consent form should 

read: “We will pay for needed medical treatment that is not paid for by your own 

insurance or the insurance of a third party under which you are covered.” 

The revised language now reads: AEATF will cover the cost of reasonable and 

appropriate medical attention for a study-related injury or illness that is not covered 

by your own insurance or insurance provided through your employer. This includes 

deductible costs and any out-of-pocket expenses, including co-payments you might 

have. The Study Director, in consultation with the on-site medical professional will 

decide if you have an illness or injury that is due to your participation in this study. 

See Appendix B, page 78. 
Tim Leighton 

Summary of changes to the Aerosol Study Protocol 

February 21, 2010 

Page 5 

d) As some possible subjects for this study may be undocumented immigrants, 

recruitment materials should more explicitly state that a valid government-issued 

form of identification is necessary for enrollment. 

The following was added to the recruitment flyer: subject must “have a valid photo 

ID.” See Subject Recruitment, Section 9 ii, page 32. 

e) The protocol provides for use of a community notification flyer. However, one 

neglected community includes persons who might be staying in the hotels where the 

study is conducted. The flyer should be revised so that it communicates the goals of 

the study and risks to that group. These flyers, in both English and Spanish, should 

also be posted in locations so that the hotel guests are likely to see it. Alternatively, 

the researchers should consider conducting the research in areas away from hotel 

guests. 

A second ‘Notice to Hotel/Motel Guests’ flyer is now added under Community 

Involvement, Section 6 D, page 18 and Appendix H, page 127. 

f) The exclusion criteria should be revised to eliminate some groups that might be at 

higher risk of physical harm but are not presently excluded. This might include 

subjects who might be immunosuppressed for a variety of reasons, those with severe 

diabetes, and those with other conditions that pose a health risk. 

Section added to protocol: (Under Exclusion Criteria, Section 9 A iii, page 35): 

• Severe diabetes 

• Immunologically suppressed (e.g. undergoing chemotherapy, transplant patients) 

Added under Subject Self-Reporting Demographic Form, Appendix D, page 107: 

Do you have any of the following conditions? 

Moderate or severe asthma, emphysema Yes  No 

Cardiovascular disease Yes  No 

Severe diabetes Yes  No 

Immunologically suppressed Yes  No 
Tim Leighton 

Summary of changes to the Aerosol Study Protocol 

February 21, 2010 

Page 6 

4. Justification for the separating monitoring activities between clusters from seven days to 

two days. 

The seven day waiting period originally arose out of applications in agricultural settings 

which could be influenced by weather conditions and other outdoor settings. 

Most antimicrobial exposure monitoring studies, including the aerosol study, will be 

conducted indoors and weather changes are not expected to have any type of impact on 

monitoring events. In addition, there are no specific parameters that would be expected to 

change significantly with time, two vs. seven days. Furthermore, based on professional 

judgment and the experience of the AEATF in mop and wipe studies, it is anticipated that 

any increase in “diversity” that may come with a seven day time span between clusters will 

have minimal to no impact on the variation estimate when compared to other factors such as 

new subjects or a new application scenario. In AEATF’s judgment, based on previous 

experience, it is much easier to sign up and retain subjects into a study with a shorter 

waiting period between enrollment and study participation. Thus, the Task Force proposes a 

minimum of two days between clusters as adequate time to get ready for monitoring the 

next cluster in more practical and cost effective manners. 

* * * 

If you have any questions, please contact me at (703) 741-5637 or via email at 

has_shah@americanchemistry.com. 

Sincerely, 

Hasmukh Shah 

Manager, AEATF 

Cc: John Carley 

 Kelly Sherman