SRC TR

- 1 3

- 050

HPV 3 TEST RULE DATA ADEQUACY REVIEW ( FINAL ): ACUTE TOXICITY AND G ENE MUTATION Propanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester , CASRN 22527

- 63

- 5 Prepared by: Justine Von Runnen , Julie Klotzbach , Teresa Many in, and William L. Richards

Defense and Environmental Solutions Division

SRC, Inc. 7502 Round Pond Road North Syracuse, NY 13212
Contract No. EP

- W

- 09

- 027 Task FG007. 3 .009
. 00 6
Submitted to: OCSPP/ OPPT/RAD (7403M) U.S. Environmental Protection Agency 1200 Pennsylvania Avenue Washington, D.C. 20460 Bryan Lobar , Project Officer David Brooks, Work Assignment Manager Amy Benson, Alternate Work Assignment Manager

July 11 , 201 3Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

i TABLE OF CONTENTS A. SOURCE MATERIAL USED FOR ASSESSMENT
................................ .............................. 1 B. EVALUATING DATA ADEQUACY
................................ ................................ ..................... 1 C. SUMMARY OF DATA ADEQUACY FOR TESTING REQUIRED BY TE ST RULE
........ 1 D . DATA FOR SIDS ENDPOINTS
................................ ................................ .............................. 3 D

- 1. Acute Toxicity
................................ ................................ ................................ .................... 3 D

- 2. Genetic Toxicity

– Gene Mutation
................................ ................................ ..................... 5 E
. DATA ADEQUACY SUMMARY TABLE
................................ ................................ .......... 11Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

1 A. SOURCE MATERIAL USED FOR ASSESSMENT This document evaluates a submission from Eastman Chemical Company (dated February 26, 2013 ; available online at http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

2009

- 0112

- 0138 as of July 8 , 201 3) received by EPA in response to the final test rule, “Testing of Certain High Production Volume Chemicals; Third Group of Chemicals” ( 76 FR 65385 , October 21, 2011 ; referred to herein as HPV 3 ; available online at http://www.regulations.gov/ #!documentDetail;D=EPA

- HQ

- OPPT

- 2009

- 0112

- 0086 as of July 8 , 2013) for p ropanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester (CASRN 22527

- 63

- 5 ) . In this submission , Eastman Chemical Company (“Eastman”) supplied data for the acute mammalian toxicity (D) and gene mutation (E1) testing required by the HPV 3 test rule. These newly submitted data are evaluated for adequacy herein. No other toxicity data are evaluated in this report. In addition to the D and E1 testing requirement s , the HPV 3 test rule requires testing with CASRN 22527

- 63

- 5 for the following toxicity endpoints: C1 (consisting of acute toxicity to fish, acute toxicity to Daphnia , and toxicity to algae te sting if log K ow < 4.2 or chronic toxicity to Daphnia and toxicity to algae testing if log K ow ≥ 4.2), E2 (chromosomal a berration or micronucleus test) and F1 (combined repeated dose toxicity study with the reproduction/developmental toxicity screening test). B. EVALUATING DATA ADEQUACY Data available for SIDS toxicity endpoints are evaluated fo r adequacy in Section D. Each study was summarized according to a standard format and assigned a Klimisch reliability code based on its scientific merit and conduct, using professional judgment. To determine whether data were adequate to satisfy SIDS end points, study details were compared to current standard test guidelines to establish whether the study met current testing requirements. Studies for a common endpoint are grouped together in the document and an endpoint adequacy assessment is provided nea r the top of the first page of the summaries for that endpoint. If the data available for the endpoint were determined to be inadequate, a basis for that conclusion is supplied. A table summarizing data availability/adequacy for each SIDS endpoint is inc luded on the last page of this document (Section E). C . SUMMARY OF DATA ADEQUACY FOR TESTING REQUIRED BY TEST RULE The test rule classifies propanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester ( CASRN 22527

- 63

- 5 ) as a Class 1 Chemical Su bstance for which testing must be conducted with material having a purity of 99% or greater. However, in a letter to EPA dated February 20, 2012 (available online at http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

2009

- 0112

- 0107 as of July 8 , 2013), Eastman proposed the use of a bulk reaction product of the following purity: 60

- 65% propanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester (CA SRN 22527

- 63

- 5), 20

- 25% 1,3

- pentanediol, 2,2,4

- trimethyl

- 1,3

- dibenzoate (CASRN 68052

- 23

- 3), and 10

- 15% 2,2,4

- trimethyl

- 1,3

- pentanediol diisobutyrate (CASRN 6846

- 50

- 0). Eastman’s rationale for the use of this mixture (the “bulk reaction product”) was bas ed on the fact that “The manufacture of benzoate ester for commercial use has been as a bulk reaction andFinal HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

2 there is no readily available process to synthesize the chemical substance by direct reaction to achieve the purity required for Class 1 substances. Eastman believes that use of the bulk reaction product which is formulated into finished products for commercial sale will more accurately represent exposure and use of the benzoate ester and avoid a costly and technically uncertain purification procedure. ” Given that the major constituent (60

- 65%) of the bulk reaction product is the test rule substance, propanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester, CASRN 22527

- 63

- 5 and the remaining two major components (CASRNs 68052

- 23

- 3 and 6846

- 50

- 0) can be considered close analogs of CASRN 22527

- 63

- 5, EPA approved Eastman’s plan to conduct aquatic and mammalian toxicity tests on this mixture via email from P. Campanella (EPA) to G. Shrum (Eastman) o n April 19, 2012 . This mixture, referred to a s “Benzoflex 1046” in the submitted study reports , can be considered a supporting substance for CASRN 22527

- 63

- 5; as such, the toxicity of Benzoflex 1046 is expected to be representative of the toxicity of CASRN 22527

- 63

- 5
. A cute mammalian toxicity and g ene mutation test results for Benzoflex 1046, supplied on February 26, 2013 by Eastman , are adequate to satisfy the D and E1 test requirement s promulgated in the final HPV 3 test rule. In a separate submission to EPA, Eastman provided the following addit ional studies of Benzoflex 1046 :  Daphnia magna Reproduction Test  Algae Growth Inhibition Test  Chromosome Aberration Test in Human Lymphocytes in vitro Data from the above tests will be evaluated for adequacy in a separate report. Data have not yet been su bmitted for the following toxicity testing requirement: combined repeated dose toxicity study with the reproduction/developmental toxicity screening test (F1 testing)
. In a letter dated December 2 1 , 2012 ( available online at http://www.regulations.gov/ #!documentDetail;D=EPA

- HQ

- OPPT

- 2009

- 0112

- 0131 as of July 11 , 2013) , Eastman provided an update on testing status . At that time, Eastman expected initiation of a defin itive combined repeated dose toxicity study with the reproduction/developmental toxicity screening test in the second quarter of 2013
.Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

3 D . DATA FOR SIDS ENDPOINTS D - 1 . Acute Toxicity Summary of endpoint: Adequate Study 1: Acute oral toxicity stud y in rats (

Sanders, 2012 ) Title: Benzoflex 1046: a cute oral toxicity in the rat

- up - and

- down procedure Th is study is: Adequate Not Adequate TEST SUBSTANCE Identity: Benzoflex 1046; supporting substance for pr opanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester (CASRN 22527

- 63

- 5) Remarks: C lear colorless liquid; Batch No. V046212201 ; Composition: 68.8% p ropanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester ( CASRN 22527

- 63

- 5), 19. 5% 1,3

- pentanediol, 2,2,4

- trimethyl

- 1,3

- dibenzoate ( CASRN 68052

- 23

- 3) , 7% 2,2,4

- trimethyl

- 1,3

- pentanediol diisobu ty rate ( CASRN 6846

- 50

- 0) , 4.2% miscellaneous esters, 0.5% volatiles, and 0.02% moisture
. 1 METHOD Method/guideline followed: OECD TG 425 Te st type: Acute Oral Toxicity

– Up

- and

- Down Procedure GLP compliant?: Yes Year study performed: 2012 Test species: Female W istar (RccHanTM:WIST) rats ; supplied by Harlan Laboratories UK Ltd., Oxon, UK; at test initiation, animals were 8

- 12 weeks of ag e and weighed 161

- 177 g. Number of animals/sex/dose : 3 females Route of administration: Oral gavage

1 The certificate of analysis provided in the study report describes the c omposition as follows: 68.8% “diester” , 19.5% “dibenzoate”, 7% “TXIB”, 4.2% miscellaneous esters, 0.5% volatiles, and 0.02% moisture. In an email dated July 3, 2013 from G. Shrum (Eastman) to J. Schaeffer (EPA), Eastman indicated that “ diester” was synonymous with p ropanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester (CASRN 22527

- 63

- 5), “dibenzoate” was synonymous with 1,3

- pentanediol, 2,2,4

- trimethyl

- 1,3

- dibenzoate (CASRN 68052

- 23

- 3), and “TXIB” was synonymous with 2,2,4

- trimethyl

- 1,3

- pentanediol diisobu t yrate (CASRN 6846

- 50

- 0).Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

4 Vehicle: None Doses/concentrations tested: 2000 mg Benzoflex 1046 /kg

- bw Statistical methods used: Not applicable Test method/conditions remarks: One fasted animal was administered undiluted test substance at a dose level of 2000 mg/kg

- bw of Benzoflex 1046 mixture . Based upon no observed mortality in this animal, two additional animals were treated sequentially at the same dose level. Sufficient time was allowed between dosing of each individual animal to confirm the survival of the previously dosed animal. Following treatment, animals were observed for mortality and systemic signs of toxicity at 0.5, 1, 2 , and 4 hours post

- dose and subsequently once daily for 14 days. Body weights were recorded prior to dosing and on days 7 and 14. Following the 14

- day observation period, all animals were sacrificed and su b jected to gross pathological examination. RESULTS Endpoint value(s): LD 50 > 2000 mg B enzoflex 1046 /kg

- bw Results remarks: No deaths, signs of systemic toxicity , or effects on body weights were observed. No abnormal findings were noted at necropsy. CONCLUSIONS The oral LD 50 for female rats was > 2000 mg Benzoflex 1046 /kg

- bw. STUDY R ELIABILITY [ 1 ] Reliable with out restrictions. REFERENCE Sanders, A. 2012
. Benzoflex 1046: Acute Oral Toxicity in the Rat

– Up

- and

- Down Procedure. Harlan Laboratories Ltd., Project No. 41201919. Sponsor: Eastman Chemical Company
. Available online a t http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2009

- 0112

- 0138 as of July 8 , 2013.Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

5 D - 2 . Genetic Toxicity

– Gene Mutation Summary of endpoint: Adequate Study 1 : Mutagenicity study in Salmonella typhimurium and Escherichia coli ( Thompson, 2012 ) Title: Benzoflex 1046: R everse mutation assay “ A mes t est” using Salmonella typhimurium and Escherichia coli Th is study is: Adequate Not Adequate TEST SUBSTANCE Identity: Benzoflex 1046; supporting substance for p ropanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester ( CASRN 22527

- 63

- 5 ) Remarks : C l ear colorless liquid; Batch No. V046212201 ; Compositio n: 68.8% p ropanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester (CASRN 22527

- 63

- 5), 19.5% 1,3

- pentanediol, 2,2,4

- trimethyl

- 1,3

- dibenzoate (CASRN 68052

- 23

- 3), 7% 2,2,4

- trimethyl

- 1,3

- pentanediol diisobutyrate (CASRN 6846

- 50

- 0), 4.2% miscellane ous esters, 0.5% volatiles, and 0.02% moisture. 1 METHOD Method/guideline followed: OECD TG 471 ; Japanese Regulatory Authorities (METI, MHLW and MAFF ) guidelines ; Method B13/14 of Commision Regulation (EC) number 440/2008; EPA (TSCA) OPPTS harmonized guidelines Test type: Bacterial Reverse Mutation Test G LP compliant?: Yes (with the exception of a nalysis for concentration, homogeneity and stability of the test item formulations ) Year study performed: 2012 Test species: Salmonella typhimurium st rains TA98, TA100, TA1535 and TA1537 and Escherichia coli strain WP2 uv r A Metabolic activation : With (rat S9 microsomal fraction) and without

1 The certificate of analysis provided in the study report describes the c om position as follows: 68.8% “diester” , 19.5% “dibenzoate”, 7% “TXIB”, 4.2% miscellaneous esters, 0.5% volatiles, and 0.02% moisture. In an email dated July 3, 2013 from G. Shrum (Eastman) to J. Schaeffer (EPA), Eastman indicated that “ diester” was synonymo us with p ropanoic acid, 2

- methyl

- , 3

- (benzoyloxy)

- 2,2,4

- trimethylpentyl ester (CASRN 22527

- 63

- 5), “dibenzoate” was synonymous with 1,3

- pentanediol, 2,2,4

- trimethyl

- 1,3

- dibenzoate (CASRN 68052

- 23

- 3), and “TXIB” was synonymous with 2,2,4

- trimethyl

- 1,3

- pentan ediol diisobu t yrate (CASRN 6846

- 50

- 0).Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

6 Concentrations tested: 0 (negative control), 0 (vehicle control ) , 50, 150, 500, 1500 and 5000 μg Benzoflex 104 6 /plate Statistical methods used: Statistical analysis of data was determined by UKEMS (Mahon et al
. , 1989 ; Analysis of data from microbial colony assays, In: Statistical Evaluation of Mutagenicity Test Data, UKEMS sub

- committee on guidelines for mutage nicity testing, Kirkland, D.J., Ed., Cambridge University Press Report, 26

- 65 ). Vehicle: Dimethyl sul f

oxide Test method/conditions remarks: Preparation of Test Items: “The test item [Benzoflex 1046 mixture] was immiscible in sterile distilled water at 50 mg/ml but was fully miscible in dimethyl sulphoxide at the same concentration in solubility checks performed in

- house. Dimethyl sulphoxide was selected as the vehicle.” [Thompson , 2012, p. 9] “The test item was accurately weighed and approximate half

- log dilutions prepared in dimethyl sulphoxide by mixing on a vortex mixer and sonication for 10 minutes at 40°C on the day of each experiment. All formulations were used within four hours of preparation and were assumed to be stable for this period. Ana lysis for concentration, homogeneity and stability of the test item formulations is not a requirement of the test guidelines and was, therefore, not determined. This is an exception with regard to GLP and has been reflected in the GLP compliance statement
. Prior to use, the solvent was dried to remove water using molecular sieves i.e. 2 mm sodium alumino

- silicate pellets with a nominal pore diameter of 4 x 10 - 4 microns.” [Thompson 2012, p. 9] A vehicle control group was treated with the solvent
. T he foll owing positive controls were used without S

- 9 mix: N

- ethy l

- N'

- nitro

- N

- nitrosoguanidine (ENNG): 2 μ g/plate for WP2 uv r A N

- ethy l

- N'

- nitro

- N

- nitrosoguanidine (ENNG): 3 μ g/plate for TA100 N

- ethy l

- N'

- nitro

- N

- nitrosoguanidine (ENNG): 5 μ g/plate for TA1535 9

- Aminoacridine (9AA): 80 μg/plate for TA1537 4

- Nitroquinoline

- 1

- oxide (4NQO): 0.2 μg/plate for TA98 The following positive controls were used in the presence of S

- 9 mix : 2

- Aminoanthracene (2AA): 1 μg/plate for TA100 2

- Aminoanthracene (2AA): 2 μg/plate for TA1535 and TA1537 2

- Aminoanthracene (2AA): 10 μg/plate for WP2 uvr A Benzo(a)pyrene (BP): 5 μg/plate for TA98Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

7 Preliminary Toxicity Test : “ In order to select appropriate dose levels for use in the main test, a preliminary assay was carried out to det ermine the toxicity of the test item [ Benzoflex 1046 mixture]
. The concentrations [of test item] tested were 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 μg/plate. The assay was performed by mixing 0.1 ml of bacterial culture (TA100 orWP2 uvrA), 0.1 ml of test item formulation, 0.5 ml of S9

- mix or phosphate buffer and 2 ml of molten, trace histidine or tryptophan supplemented, top agar and overlaying onto sterile plates of Vogel

Bonner Minimal agar (30 ml/plate). Ten concentrations of the test i tem and a vehicle control (dimethyl sulphoxide) were tested. In addition, 0.1 ml of the maximum concentration of the test item and 2 ml of molten, trace histidine or tryptophan supplemented, top agar were overlaid onto a sterile nutrient agar plate in ord er to assess the sterility of the test item. After approximately 48 hours incubation at 37°C the plates were assessed for numbers of revertant colonies using a Domino colony counter and examined for effects on the growth of the bacterial background lawn.” [Thompson 2012, p. 11] Mutation Test

- Experiment 1 (Range

- finding Test) : “ Five concentrations of the test item (50, 150, 500, 1500 and 5000 μ g/plate) were assayed in triplicate against each tester strain, using the direct plate incorporation method.” [T hompson 2012, p. 12]

“ Measured aliquots (0.1 ml) of one of the bacterial cultures were dispensed into sets of test tubes followed by 2 ml of molten, trace histidine or tryptophan supplemented, top agar, 0.1 ml of the vehicle, test item formulation or posi tive control and either 0.5 ml of S9

- mix or phosphate buffer. The contents of each test tube were mixed and equally distributed onto the surface of Vogel

Bonner Minimal agar plates (one tube per plate). This procedure was repeated, in triplicate, for eac h bacterial strain and for each concentration of test item both with and without S9

- mix.” [Thompson 2012, p. 12]

“ All of the plates were incubated at 37°C for approximately 48 hours and the frequency of revertant colonies assessed using a Domino colony co unter.” [Thompson 2012, p. 12] Mutation Test

- Experiment 2 (Main Test) : “ The second experiment was performed using fresh bacterial cultures, test item and control solutions. The test item dose range was the same as the range

- finding test (50 to 5000 μ g/ plate).” [Thompson 2012, p. 12]

“ As it is good scientific practice to alter one condition in the replicate assay, the exposure condition was changed from plate incorporation to pre

- incubation. The test item formulations and vehicle control were therefore dosed as follows: Measured aliquots (0.1 ml) of one of the bacterial cultures were dispensed into sets of test tubes followed by 0.5 ml of S9

- mix or phosphate buffer and 0.1 ml of the vehicle or test item formulation and incubated for 20 minutes at 37° C with shaking at approximately 130 rpm prior to the addition of 2 ml of molten, trace histidine or tryptophan supplemented, top agar. The contents of the tube were then mixed and equally distributed on the surface of Vogel

- Bonner Minimal agar plates (one t ube per plate). This procedure was repeated, in triplicate, for eachFinal HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

8 bacterial strain and for each concentration of test item both with and without S9

- mix. The positive and untreated controls were dosed using the standard plate incorporation method descr ibed [ above ]
. ” [Thompson 2012, p. 1 2 ]

“ All of the plates were incubated at 37°C for approximately 48 hours and the frequency of revertant colonies assessed using a Domino colony counter.” [Thompson 2012, p. 13] Acceptance Criteria: According to the study report, t he following criteria must be met for the assay to be considered valid:

“ All bacterial strains must have demonstrated the required characteristics as determined by their respective strain checks
... All tester strain cultures should exhibit a characteristic number of spontaneous revertants per plate in the vehicle and untreated controls
... All tester strain cultures should be in the range of 0.9 to 9 x 10 9 bacteria per ml. Diagnostic mutagens (positive control chemicals) must be included to de monstrate both the intrinsic sensitivity of the tester strains to mutagen exposure and the integrity of the S9

- mix. All of the positive control chemicals used in the study should induce marked increases in the frequency of revertant colonies, both with or without metabolic activation
... There should be a minimum of four non

- toxic test item dose levels. There should be no evidence of excessive contamination. ” [Thompson 2012, p. 13] Evaluation Criteria: “There are several criteria for determining a positive result. Any, one, or all of the following can be used to determine the overall result of the study: 1. A dose

- related increase in mutant frequency over the dose range tested
... . 2. A reproducible increase at one or more concentrations. 3. Biological relevance agai nst in

- house historical control ranges. 4. Statistical analysis of data as determined by UKEMS
... 5. Fold increase greater than two times the concurrent solvent control for any tester strain (especially if accompanied by an out

- of

- historical range response).” [Th ompson , 2012, p. 13

- 14] “A test item will be considered non

- mutagenic (negative) in the test system if the above criteria are not met.” [Thompson 2012, p. 14] “Although most experiments will give clear positive or negative results, in some instances the data generated will prohibit making a definite judgement about test item activity. Results of this type will be reported as equivocal.” [Thompson 2012, p. 14] RESULTS Genotoxicity result: Negative with and without metabolic activation Cytotoxic conce ntration (if applicable): > 5000 μg Benzoflex 1046 /plate , with and without S

9 mi xFinal HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

9 Results remarks: Preliminary Toxicity Test: “ The test item was non

- toxic to the strains of bacteria used (TA100 and WP2 uvr A ). The test item formulation and S9

- mix us ed in this experiment were both shown to be sterile.” [Thompson 2012, p. 15] Mutation Test s : “Prior to use, the master strains were checked for characteristics, viability and spontaneous reversion rate (all were found to be satisfactory). These data are not given in the report. The amino acid supplemented top agar and the S 9

- mix used in both experiments was shown to be sterile.” [Thompson 2012, p. 15] Results for the negative ( untreated ) and vehicle controls were acceptable. “The test item caused no v isible reduction in the growth of the bacterial background lawn at any dose level and was, therefore, tested up to the maximum recommended dose level of 5000 μ g/plate. A test item precipitate (oily in appearance) was noted at and above 1500 μ g/plate, this observation did not prevent the scoring of revertant colonies.” [Thompson 2012, p. 16] “No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation or [using either] exposure method.” [Thompson 2012, p. 16] “All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies thus confirming the activity of the S9

- mix and the sen sitivity of the bacterial strains.” [Thompson 2012, p. 16] CONCLUSIONS The test chemical , Benzoflex 1046 , d id not induce gene mutations in bacteria either with or without metabolic activation. STUDY RELIABILITY [ 2 ] Reliable with restrictions. The seco nd of two assays, which used the preincubation method, did not include positive or untreated controls test ed by that method. Instead, “[t] he positive and untreated controls [ run concurrently with cells exposed to the test substance using the preincubation method in Experiment 2 ] were dosed using the standard plate incorporation method
... ” [Thompson 2012, p. 12]Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

10 REFERENCE Thompson, P.W. 2012. Benzoflex 1046: Reverse Mutation Assay “Ames Test” Using Salmonella typhimurium and Escherichia coli . Harlan Lab oratories Ltd., Project No. 41201917. Sponsor: Eastman Chemical Company. Available online at http://www.regulations.gov /#!documentDetail;D=EPA

- HQ

- OPPT

- 2009

- 0112

- 013 8 as of July 8 , 2013. Text in quotes is taken directly from Thompson ( 2012 )
.Final HPV 3 Chemical Data Adequacy Review CASRN 22527 - 63 - 5

11 E . DATA ADEQUACY SUMMARY TABLE

Endpoint Study Information Available Acceptable Data Gap Aquatic Effects Acute Toxicity to Fish No

– – 1 Acute Toxicity to Aquatic Inve rtebrates No

– – 1 Chronic Toxicity to Aquatic Invertebrates Yes 2

– TBD Toxicity to Aquatic Plants Yes 2

– TBD Health Effects

Acute Toxicity Yes Yes No Repeated

- Dose Toxicity No 3

– Yes Gene Mutation Yes Yes No Chromosomal Aberrations Yes 2

– TBD Rep roductive Toxicity No 3

– Yes Developmental Toxicity No 3

– Yes
1 Acute toxicity to fish and acute toxicity to aquatic invertebrate testing are not required if the measured log K ow of CASRN 22527

- 63

- 5 is < 4.2. 2 Data were received from Eastman via email and will be evaluated for adequacy in a separate report. 3 According to Eastman’s letter to EPA dated December 2 1 , 2012, Eastman expected initiation of a definitive combined repeated dose toxicity study with the reproduction/developmental toxicity screenin g test in the second quarter of 2013. TBD = To be determined