211(b) Toxicology Research Group Gasoline Tame VC: 

A 13- Week Whole Body Toxicity Study in Rats with Neurotoxicity 

Assessments and 4-Week In Vivo Genotoxicity and 

ImmunotoxicityAssessments 

Reviewer Checklist 

1 10/16/2009 

Inhalation Toxicity Report 

Comments Action/Response Page 

211(b) Research Group QA/QC Reviewer 

The following item requires consideration: 

2. Compliance Statement: Since it is the sponsor’s 

responsibility to maintain the method of synthesis, 

fabrication, or derivation of the test fuel, and this has not 

been completed, it must be included in the sponsor’s 

compliance statement. 

Added to Compliance Statement. Needs to be 

updated with latest sentence used in other 

reports. (information was not available 

during study, but is now maintained with 

sponsor) 

Compliance statement up-dated as indicated. 

4. Page 4, Quality Assurance Statement: Since HLS has 

inspected all of the subcontractors (except CDC) these 

facility inspections should be listed on the QA statement for 

this study (Immumotox and BioReliance periodic 

inspections). 

Our inspections of the Subcontractors added 

to our included QA Statements. I don’t see 

inspections of these two facilities. Those 

listed are for work done at HLS. 

Facility inspections are listed on the HLS QA 

statements contained within each sub-contract 

report.

6. Page 7, last sentences, first paragraph: There was a 

variation noted during Exposure 13. It is noted as 

“suspicious” on the table, but no mention of it occurs 

anywhere. Unless it is known to be in error, the variation 

should be reported. 

We don’t find any ‘suspect’ values noted in 

the CMR table in Appendix A for Exposure 

13 or any other exposure day. Sorry, this was 

in the analytical report. Appendix S... 

Exposure 13, footnote indicates suspect 

values. Not noted whether included in mean 

or not. Was it, and should it have been? 

The sample is included in the mean in the 

Summary Table. The word “suspect” is 

removed from the report (Exposure 13 table) 

to avoid confusion. 

11. Page 47, Chamber monitoring table: Group 4, 

nominal concentration standard deviation is 661 in other 

tables. 

Corrected. I cannot verify. Starting on page 

49 of the text of the report (Results and 

Discussion), pages from ETOH are included. 

This continues until page 62, so I can’t verify 

changes through item #17. In scanning 

through the appendices, it has come to our 

attention that not only the text of the report, 

but several of the Appendices have Ethanol 

Tables. The TAME Micronucleus and the 

Sister Chromatid Exchange reports contain 

tables from the Ethanol report. At least the 

headings of the tables indicate ethanol. 

We assume/hope that this is just an error in 

collecting all of the sections of data for the 

reports, but request that HLS QA check to 

make sure all appropriate tables, appendices 

and text appear in the next version of this 
211(b) Toxicology Research Group Gasoline Tame VC: 

A 13- Week Whole Body Toxicity Study in Rats with Neurotoxicity 

Assessments and 4-Week In Vivo Genotoxicity and 

ImmunotoxicityAssessments 

Reviewer Checklist 

2 10/16/2009 

Inhalation Toxicity Report 

Comments Action/Response Page 

report. 

Report originals checked and correct R/D is 

present; incorrect (00-6127) R/D does 

however appear to be in the PDF file 

submitted for 2nd draft. In-life tables 

presented with micronucleus and SCE reports 

were the correct files, though it appears that 

an error was made in the entry of the TASC 

protocols in as much as the wrong test 

substance was listed in the title. The study 

title was subsequently corrected and the tables 

have been reprinted. The remainder of the 

report was checked and all pages presented 

are for the indicated study (TAME). 

12. Page 48, last paragraph: data were consistent from 

week-to-week except for exposure 13, where there should be 

some mention of the “suspicious” values for the group three 

chamber. 

We don’t find any ‘suspect’ values noted in 

the CMR table in Appendix A for Exposure 

13 or any other exposure day. 

13. Page 50, Motor Activity: The statement that the 

subsequent statistical evaluation of the data did not confirm 

this initial determination, made in the last sentence is not 

altogether true. There were some statistically significant 

findings in the 13th week data. They should be discussed 

here. 

The Mark Nicolich text has been added here 

although the conclusion of no effect has not 

changed.

14. The draft report, on page 51, section 3.8.3 Clinical 

Chemistry discusses the parameters that were statistically 

significant. The dosage levels should be added to this 

section to clarify these results. (alkaline phosphatase, males 

20000 mg/m3

; chloride, females 10000 and 20000mg/m3) 

Exposure levels have been added. 

15. The draft report, on page 51, section 3.8.3 Clinical 

Chemistry discusses the terminal sacrifice results. In the 

third sentence of this section, male animals in the 

20,000mg/m3

 level should be added and the fourth sentence 

should add in the 10,000 and 20,000 mg/m3

 levels to clarify 

the results. 

Exposure levels have been added.

16. The draft report text on page 52, section 3.9 Organ 

Weights and Brain Measurements should be reviewed to 

clarify the discussion on liver weights. Only three liver 

weights were statistically significant: Male relative to body 

weight, 20,000mg/m3

, terminal sacrifice; female, absolute 

weight, 20,000mg/m3

 recovery sacrifice; male relative to 

body weight 20,000mg/m3

 recovery sacrifice. 

The current text is adequately descriptive and 

the reader may review the tables for more 

details for these no effects differences.

17. Page 52, Macroscopic Pathology: It may be helpful to 

indicate that 6 of 20, meaning 3 main study animals and 3 

recovery animals, exposed to 20,000.... 

Added to this text. 
211(b) Toxicology Research Group Gasoline Tame VC: 

A 13- Week Whole Body Toxicity Study in Rats with Neurotoxicity 

Assessments and 4-Week In Vivo Genotoxicity and 

ImmunotoxicityAssessments 

Reviewer Checklist 

3 10/16/2009 

Inhalation Toxicity Report 

Comments Action/Response Page 

20. Table 12 and Appendix L do not contain the Sponsor 

Study Number in the heading. The individual pages in 

Appendix L and Table 12 are not labeled as such. 

Headings will remain as is. The computer 

program does not allow for addition of a 

sponsor study number. Table 12.... 01-6128 

on page 1 ... should be 00-6128 

Typographical error in the heading of the 

preface page has been corrected.
211(b) Toxicology Research Group Gasoline Tame VC: 

A 13- Week Whole Body Toxicity Study in Rats with Neurotoxicity 

Assessments and 4-Week In Vivo Genotoxicity and 

ImmunotoxicityAssessments 

Reviewer Checklist 

4 10/16/2009 

GFAP SATELLITE REPORT 

Comments Action/Response Page 

GFAP LEVELS IN SPECIFIC RAT BRAIN AREAS FOLLOWING A 13-WK WHOLE BODY INHALATION 

EXPOSURE TO BASELINE GASOLINE VAPOR 

Peer Reviewer – Schlesinger

Peer Reviewer - Goldsworthy 

Pituitary GFAP levels could only be detected in less than 

25% of samples; due to the paucity of data and inability to 

make any conclusions from this region of the brain, this data 

should be omitted from Tables and noted in text that analysis 

of pituitary could not be adequately performed. 

WG agrees with the reviewer’s comments. 

Sparse pituitary data has been deleted from 

Tables 5 and 6, but retention of individual 

values in Table 7, primarily because this 

series of studies is one of the first in which 

GFAP levels in pituitary have been measured. 

Although other 211(b) reports in the series 

show somewhat better recovery of pituitary 

data, this study has no usable data for females 

in control, low or high doses and only 1 entry 

at mid-dose; data for males is only slightly 

better. To include this limited data in 

summary tables (5&6) gives an erroneous 

impression that sufficient animals/group 

responded. Keeping individual entries (Table 

7) provides an idea of the level of GFAP 

likely to be present in this small organ, 

providing a starting point for improvement of 

assay methods. Results and conclusions can 

also be revised to indicate that GFAP was not 

increased in 8 areas of the brain, pituitary 

excluded.

Done. 

Note that text, page 18, states that ‘half of pituitaries’ gave 

results below level of sensitivity. This is incorrect according 

to Table 7, where the majority (9/40) of animals were shown 

to be below limits of detection. 

Since the pituitary entry in Table 7 is retained, 

the text on p. 18 has been corrected 

The footnote content on Table 7 should appear 

on Page 17 or 18 to clarify. 

Done