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## 2012 DEC -7 A;.i 10; 42 3676 Davis Road N

.W . " Dover, Ohio 44622 " (330) 343-7711 " 1-800-321-8805 " Fax: (330) 364-9626 November 30, 2012 Document Control Office (DCO) (7407M) Office of Pollution Prevention and Toxics (OPPT) Environmental Protection Agency 1200 Pennsylvania Avenue, NW Washington, DC 20460 ATTN : TSCA Section 4 40 CFR799.5089

; Docket ID Number EPA-HQ-OPPT-2009-0112 Dover Chemical Study Plan for CAS# 72162-15-3 Towhom

it may concern: IIIIII IIIII IIIII IIIII IIIII IIIII IIIII IIIII IIIII IIIII IIIII IIII IIII 4 0 1 3 0 0 0 0 0 1 7

Dover Chemical Corporation is requesting an extension for the test completion of several studies on (CAS# 72162-15-3) based on the EPA's HPV3 TEST PLANAND DATA ADEQUACY REVIEW (Contract No. EP-W-09-027) and our difficulties in scheduling lab space and scheduling the testing. We request the following extensions: Chronic Toxici ty to Da hnia For Chronic Toxicity to Daphnia we request an extension to the 3`d quarter of 2013. The chronic daphnia assay will be initiated following completion of the water solubility assessment . Reoroductive Toxicity Developmental Toxici For the Combined Repeat Dose Toxicity with Reproduction/Developmental Tox Screening we request an extension to the 4th quarter of 2013. The reproduction/developmental/repeated dose study will be initiated in 2nd quarter of 2013. All physical/chemical testing in the first quarter of2013, and expects results no later than the 2ndquarter 2013. Should you have any questions, please contact me.



## CONTAINS NOCB[

## " Corpo~r~ion

" //WL1rOf14Y1CY~YWK . 3678 Davis Road N

.W . " Dover, Ohio 44822 " (330) 343-7711 " 1-800-321-8805 " Fax: (330) 3648828 Sincerely,

Tim Nowicki Manager Product Compliance Dover Chemical Corporation 330-365-3610 tnowicki(~a,doverchem

.com Attachments HPV3 TEST PLAN ANDDATA ADEQUACY REVIEW 1-Decene, sulfurized, CASRN 72162-15-3 Proposed Test Plan; Testing of Certain High Production Volume Chemicals; Third Group of Chemicals. Federal Register 76 (204), 65385-65410, October 21, 2011 . 1-Decene, sulfurized (CAS No. 72162-15-3)p. 3

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## " Cor~poilration

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# . . .~. . . .~~. . 3676 Davis Road N

.W .- Dover . Ohio 44622 - (330) 343-7711 - 1-800-321-6605 - Fax : (330) 364-8626 November 30, 2012 Document Control Office (7407 M) Office of Pollution Prevention and Toxics (OPPT) Environmental Protection Agency EPA East Building, RoomE6428 (202-564-8930) 1201 Constitution Avenue, NW Washington, DC 20460 Attention : TSCA Section 4 Docket ID Number EPA-HO-OPPT-2009-0112 To Whom

It May Concern: RE : Proposed Test Plan; Testing of Certain High Production Volume Chemicals ; Third Group of Chemicals. Federal Register 76 (204), 65385-65410, October 21, 2011 . 1-Decene, sulfurized (CAS No. 72162-15-3) Dover Chemical Corporation submits this test plan to satisfy the testing requirements of the subject test rule for 1-Decene, sulfurized (CAS # 72162-15-3) . Dover Chemical Corporation agrees to conduct the following studies or provide read across data on appropriate compounds that can be bridged to satisfy one or more of the requirements in compliance with the test rule and according to current OPPTS Harmonized Guidelines. Test Facilitv Dover Chemical Corporation has entered a contract to performthe studies outlined in this test plan at the following testing laboratory; WIL Research Europe Hambakenwetering 7 5231 DD's-Hertogenbosch The Netherlands Phone: +31 (0) 73 640 6700 Fax: +31 (0) 73 640 6799 www .wilresearch.com



## CONTAINS NOCB!Test Material

Identification: Thetest itemwill be supplied by the Sponsor with details of purity and known properties if known. The integrity of data supplied relating to the identification, purity and stability of the test itemwill be the responsibility of the Sponsor. Storage: The test itemwill be stored at roomtemperature, in the dark, unless otherwise specified by the Sponsor . Analysis: Details of identification of the test itemwill be supplied by the study Sponsor . The test item formulations will be analyzed for concentration and stability by WIL Research Europe, 'sHertogenbosch, the Netherlands . Phys ical/Chemical Properties To Fulfill Testing Requirements A2 to 5 Boiling point : ECA

.2 / OECD 103/ OPPTS 830 .7220 Vapor Pressure: ASTM E 1782-08 (thermal analysis) N-Octanol/water partition coefficient (log 10 basis) or IogKoW by appropriate method as specified as Method A, 40 CFR 799.6755 (shake flask) ; Method B, ASTM E 1147-92 (reapproved 2005) (liquid chromatography) ; or Method C, 40 CFR 799.6756 (generator column) . Atest to determine the octanol/water partition coefficient is not required when the estimated log Kow (the log of the octanol/water partitioning coefficient) of the chemical is < 0 . Water solubility: By appropriate test method as specified as Method A, ASTM E 1148-02 (reapproved 2008) (shake flask) ; Method B, 40 CFR 799.6784 (shake flask) ; Method C, 40 CFR 799.6784 (column elution) ; or Method D, 40 CFR 799.6786 (generator column) . Aquatic Toxicitv-to_Fulfill Testing Requirement C1 as a Test Group 2 Substance Based upon the estimated log Kow (the log of the octanol/water partitioning coefficient) of 1- Decene, sulfurized being z 4 .2. We intend to perform aquatic toxicity Test Group 2 as detailed below

; Chronic Toxicity to Daphnia: ASTM E 1193-97. Mammalian Toxicity-Repeated Dose/Reproduction/Developmental to Fulfill Testing Requirement F1 Combined repeated dose toxicity study with reproduction/developmental toxicity screening test, 40 CFR 799.9365/OPPTS 870 .3650/OECD422Initiation of the required testing is subject to Environmental Protection Agency's guidance on the

recommendations offered in this letter. We welcome the opportunity to discuss this test plan in a meeting or conference call at the earliest opportunity so we can begin testing as quickly as possible. Please contact me directly at 330-365-3610 for questions or additional information. Sincerely,

TimNowicki Manager Product Compliance Dover Chemical Corporation tnowicki(cbdoverchem-comp. 6 ' -

G D15 SRC TR-12-034

HPV3 TEST PLANAND DATA ADEQUACY REVIEW 1-Decene, sulfurized, CASRN 72162-15-3

Prepared by: Teresa Manyin, ChristopherJanson, WilliamL . Richards, and Gary Diamond Chemical, Biological and Environmental Center

SRC, Inc. 7502 Round Pond Road North Syracuse, NY 13212 Contract No. EP-W-09-027 Task FG007.2.009.006 Submitted to: OCSPP/OPPT/RAD (7403M) U.S . Environmental Protection Agency 1200 Pennsylvania Avenue Washington, D

.C. 20460 Debra Milligan, Project Officer Meena Sonawane, Work Assignment Manager David Brooks, Alternate Work Assignment Manager

March 14, 2012



## CONTAINS "d0 CB!Final HPV3 Test Plan and Data Adequacv Review CASRN72162-15-3 TABLE OF CONTENTS A . INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 B . SUMMARY OF DATA ADEQUACY FORTESTING REQUIRED BYTEST RULE . . . . . . . . 2 C . ROBUST SUMMARIES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 C-1 . C-2 . C-3 . C-4 . C-5 . C-6 . C-7. C-8 . C-9 . Acute Toxicity to Fish. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 Acute Toxicity to Daphnia. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 Chronic Toxicity to Daphnia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Toxicity to Plants (Algae) . .. . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 AcuteToxicity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 Repeated-Dose Toxicity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 Genetic Toxicity- Bacterial Reverse Mutation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Genetic Toxicity- Chromosomal Aberrations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 Reproductive Toxicity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60 C-10 . Developmental Toxicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 D . DATA ADEQUACY SUMMARYTABLE. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

HPV3 Test Plan and Data Adequacy Review : 1-Decene, sulfurized, CASRN72162-15-3 A . INTRODUCTION The test rule, "Testing of Certain High Production Volume Chemicals; Third Group of Chemicals" (76 FR 65385, October 21, 2011), requires the following toxicity tests for 1-decene, sulfurized (CASRN 72162-15-3) : C 1 -Aquatic Toxicity D- Mammalian Acute Toxicity El -Bacterial Reverse Mutation E2 - Chromosomal Damage (Aberration or Micronucleus Test) Fl - Mammalian Repeated Dose/Reproduction/Developmental Toxicity For the C

1 testing requirement, thetest rule requires the use of Test Group 1 if log Kaw < 4 .2, whereas Test Group 2 is required if log Kaw >- 4.2. Given that the estimated log Kow of 1-decene, sulfurized is > 6

.0, the use ofTest Group 2 is required for this chemical substance. Test Group 2 consists of ChronicToxicity to Daphnia and Toxicity to Plants (Algae) . To address the required endpoints, Dover Chemical Corporation (referred to as Dover hereafter) submitted a test plan,' which utilizes data frommembers of the Alkyl Sulfides HPVCategory in an analog approach. The submitter proposes the use ofdata for the following three analogs for some ofthe required toxicity endpoints: 1-Propene, 2-methyl-, sulfurized, CASRN6841-50-2 Pentene, 2,4,4-trimethyl-, sulfurized, CASRN ;¢8515-88-8 Alkenes, C15-18 alpha-, sulfurized, CASRN 67762-55-4 The proposed analog chemicals were classified as the "Alkyl Polysulfides" Subcategory in EPA's Hazard Characterization of the Alkyl Sulfides Category.Z EPAaccepts the use ofthe proposed analogs because they are structurally similar to the test rule chemical substance3 with similar functional groups andall four chemicals share similar physicochemical characteristics,4 including high partition coefficient (log Kow), low water

' Letter dated February 20, 2012. Z Screening-Level Hazard Characterization: Alkyl Sulfides Category, dated September 2009; available at http://www

.e ap~ . ov/chemrtk/hpvis/hazchar/Cateaorv Alkvl%20Sulfides Seot2009.ndf. 3 Dover enoneously states that the analogs and test rule chemical substance are all composed of highly branched alkanes (on pp. 3 and 9 of Dover's test plan) . Dover's test plan indicates that the test rule chemical "is manufactured by reacting molten sulfur with 1-decene" and that the "decene feedstock is a mono-unsaturated highly branched C10 hydrocarbon" (on p

. 1 of Dover's test plan) . However, according to Chemical Abstract Service nomenclature conventions, both the 1-decene feedstock and the test rule chemical, 1-decene, sulfurized (CASRN 72162-15-3), are non-branched. Ofthe three chemicals that Dover proposes as analogs ofthe test rule chemical, only CASRNs 68511-50-2 and 68515-88-8 are composed of highly branched alkanes (as shown in Figure 1 of the initial test plan for theAlkyl Sulfide Category, submitted by the Health, Environmental, and Regulatory Task Groupof the Chemical Manufacturers Association Petroleum Additives Panel under the HPVChallenge Programand available at htto://www

.eua .flov/huv/pubs/summaries/alkvlsul/c12549to .odf ) . Although only two ofthe four chemicals of the Dover test plan (the test rule chemical and three analogs) are composed of highly branched alkanes, all four chemicals possess sufficient structural similarities to allowread across of toxicity values. 4Values estimated using EPISuite (p. 4 of Dover's test plan) .Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

solubility, low

vapor pressure and high boiling point . Additionally, the molecular weights of the analogs and test rule chemical are generally similar. B . SUMMARY OF DATA ADEQUACY FOR TESTING REQUIRED BY TEST RULE No experimental toxicity data are available for the test rule chemical . Dover proposes a readacross ("bridging") strategy using analog data, as well as modeled data, to satisfy the test-rulerequired endpoints. For aquatic toxicity endpoints, Dover suggests that testing is "not applicable based upon physicochemical properties of the substance, primarily low watersolubility which makes certain tests not feasible" . Dover agrees to conduct a test for only one ofthe toxicity endpoints: acute oral toxicity. Asummaryof EPA's response/assessment for each endpoint follows: 1 . Chronic Toxicity to Daphnia No experimental data are available for the test rule chemical or analogs. Although Dover provides ECOSAR-estimated chronic daphnid toxicity values for "plausible structures of lower molecular weight decyl sulfides", modeled data are not acceptable as a substitute for measured data. Furthermore, low water solubility does not preclude the conduct of a chronic daphnid test . The test rule guideline, ASTM E 1193-97 (Reapproved 2004) provides guidance for useof test substances that are slightly soluble in water. Therefore, the testing requirement for chronic toxicity to Daphnia is maintained. 2. Toxicity to Plants (Algae) Adequate data are available fromthe analog CASRN68511-50-2 to satisfy this testing requirement. 3. Acute Oral Toxicity (Rat) Adequate data are available fromthe analogs CASRNs 68511-50-2 and 68515-88-8 to satisfy this testing requirement . Although Dover proposes to conduct an acuteoral toxicity test on CASRN 72162-15-3, such testing is unnecessary because adequate analog data are available for this endpoint . 4 . Bacterial Reverse Mutation Adequate data are available fromthe analogs CASRNs 68511-50-2, 68515-88-8, and 67762-55-4 to satisfy this testing requirement . 5 . Chromosomal Aberration Adequate data are available fromthe analogs CASRNs 68511-50-2 and 68515-88-8 to satisfy this testing requirement . Data for the C12-C16 alkyl sulfide substance were notFinal HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3 considered for satisfaction of the endpoint because nojustification was provided for use of this analog. 6 . Reneated Dose/Renroduction/Develonmental Adequate data are available fromdermal studies of the analogs CASRNs 68511-50-2 and 68515-88-8 and an inhalation study of the analog CASRN 68515-88-8 to satisfy the repeated dose testing requirement . EPAcomments on the initial test plan for the Alkyl Sulfide Categorys stated that "based on the use ofthese materials (lubricant additives)" . . . "the most appropriate routes ofexposure in terms of relevance to humans appear to be either the inhalation or dermal routes." No analog data are available for the reproduction/developmental toxicity endpoint. The data from mammalian toxicity tests (e.g., repeated-dose toxicity tests) cannot be considered to constitute a substitute or alternative for reproduction/developmental toxicity testing . In general, the potential for reproduction/developmental toxicity is not inherently apparent fromother types of mammalian toxicity testing. Therefore, the testing requirement for reproduction/developmental toxicity testing is maintained. Giventhat adequate data are available for the repeated dose toxicity endpoint, the submitter may satisfy the Fl testing requirement by conducting 40 CFR 799 .9355 (Reproduction/Developmental Toxicity Screening Test) . Toxicity data presented below for the analog chemicals were obtained fromthe Final Submission for the Alkyl Sulfides Category, submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group ofthe American Chemistry Council's Petroleum

Additives Panel on May 19, 2005.6 Each available toxicity study is summarized in Section C

. Using standard testing guidelines and professional judgment as guides, each study was assigned a Klimisch reliability code. Atable summarizing the data adequacy for each toxicity endpoint is provided on the last page of this document (Section D) .

5 EPA comments on the initial test plan for theAlkyl Sulfides Category are available at

ml t "II w a ov/ol2pt/chemrtk/t)ubs/summaries/alkylsul/cl2549ct .pd h*e firialseuMssion of robust summaries for the Alkyl Sulfide Category is available at t http://www

.eoa .eov/huv/uubs/summaries/alkvlsul/cl2549rr2 .ndf.Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

C . ROBUST SUMMARIES C-1 . Acute Toxicity to Fish Summary of endpoint : Adequate Study 1 : Acute toxicity study ofCASRN68511-50-2 in fathead minnow (Ward, 1993) Title: Acute Toxicity of the Water Accommodated Fractions (WAFs) of CMA613 to the Fathead Minnow, Pimephales promelas This study is: ®

Adequate 0

NotAdequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfurized, CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: US EPAToxic Substances Control Act Test Guideline #797 .1400 (1985), OECDTG 203, Fish AcuteToxicity Test (1984) Test type: Fish acute toxicity test (WAY static renewal) GLPcompliant?: Yes Year study was performed: 1993 Test species: Fathead minnow (Pimephales promelas) Analytical monitoring: Yes; total organic carbon (TOC) measurements ofinitial test solutions and control (0-hour) and after one dayon test (24-h) before daily renewal of fresh test solution. Exposure duration: 96 hours Concentrations tested: 0, 100, 300, or 1000 mg/L (nominal loading rates) Test method/conditions remarks : Test Organisms: source - Aquatic Research Organisms, Hampton, New

Hampshire; age -juvenile; total length - 25 mm average (longest fish not more than twice the shortest fish) ; wet weight -0.1 g average (no range reported) . Loading: < 0 .5 g biomass/L; pretreatment : none, fish held for a minimum of 14 days before testing. No feeding occurred during thetest.Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Test System

: Individual WAFs were prepared for each test level and renewed daily. A measured weight of test material was added to a measured volume of dilution water (30 L) in a glass vessel and stirred for 24 hours. Stirring was accomplished using a Telfon-coated magnetic stir bar. Mixing speed was adjusted such that a vortex formed between 30 to 50% of the distance to the bottom

. Following the mixing period, the test solutions were allowed to stand for 1 hour before the water phase was removed . To avoid removing test material fromthe surface or bottom, a siphon was placed in the mixing vessel prior to addition of water and test substance with the lower end 1-2 inches off the bottom . The siphoned water phase (i .e., WAF) was used in the aquatic toxicity test . About 80%of the solution in each test level was renewed daily after 24, 48, and 72 hours. Two 15-L replicates were used per treatment, and 10 fish were used per replicate (for a total of 20 per treatment) . Test vessels were loosely covered to reduce entry of dust, etc . Dilution Water: Filtered well water collected at Hampton, New Hampshire and adjusted to the appropriate hardness of 176 mg/L as CaC03 . The water was passed through activated carbon, a particle filter, and then an ultraviolet sterilizer, and then it was stored in a polyethylene tank where it was aerated. The water was characterized as moderately hard water. Alkalinity was not reported. Dissolved oxygen: 7.1 to 8.7 mg/L; pH : 7.8 to 8 .5; conductivity: 570to 650 pmhos/cm

; temperature: 21 .4 to 22.8 °C . TOC levels were between 2 to 3 mg/L in the control, 3 mg/L at 100 mg/L test level, between 3 to 4 mg/L at 300 mg/L test level, and 5 mg/L at 1000 mg/L test concentration level . Test Levels: Control, 100, 300, & 1000 mg/L WAF loading rates. Calculation of LLscs: Statistical analysis of survival data was notwarranted. Test Substance: No undissolved test material was seen on the surface ofthe test vessels during the entire aquatic toxicity test. Reference Substance: No RESULTS Endpoint value(s) : 96-h LLso > 1000 mg/L (nominal) ; 96-h LOEC = 1000 mg/L (nominal) basedon lethargy and erratic swimming; 96-h NOEC = 300mg/L (nominal) Results remarks : Test Findings: No mortality was observed in all treatments and the control throughout the entire test and no signs oftoxicity were noted in all treatments throughout 72 hours. At 96 hours, a1120fish in the 1000 mg/L test level were lethargic and exhibited erratic swimming, but no signs of toxicity were observed in the lower test levels and control . TOCmeasurements at 1000 mg/L were 5 mg/L compared to 2 to 3 mg/L in the control . The TOC in dilution water at the beginning and endof the test was greater than 2 mg/L rather than < 2 mg/L. It could not be verified that water samples were passed through a 0 .45 micronFinal HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

filter prior to TOCanalysis, that the test vessels were covered, or that the continuous temperature measurement was made in a control vessel during the study . EPAdoes notconsider the loading rate as the no-effect concentration when the concentration exceeds the water solubility ofthe substance. Assuming that the exposure concentration in the WAF is the water solubility limit (saturation) for 1-propene, 2-methyl-, sulfurized, the no-effect concentration for lethality would be approximately 2.7 mg/L. (No lethal effects were observed at saturation.) CONCLUSIONS No mortality was observed in all treatments and the control throughout the entire test and no signs oftoxicity were noted in all treatments throughout 72 hours. At 96 hours, a1120fish in the 1000 mg/L test level were lethargic and exhibited erratic swimming, but no signs oftoxicity were observed in the lower test levels and control . STUDY RELIABILITY [2] Reliable with restrictions. TheTOCin dilution water at the beginning and end ofthe test was greater than 2 mg/L rather than < 2 mg/L. The pH range extended above 8.0. It could not be verified that water samples were passed through a 0 .45 micron filter prior to TOCanalysis, that the test vessels were covered, or that the continuous temperature measurement was made in a control vessel during the study. REFERENCE Ward, T.J . 1993. Acute Toxicity ofthe WaterAccommodated Fractions (WAFs) of CMA 613 to the Fathead Minnow, Pimephales promelas . T.R. Wilbury Study#9176-CMA/ESI-613. As summarized in the Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's PetroleumAdditives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 2: Acute toxicity study ofCASRN68511-50-2 in sheepshead minnow (Nicholson, 1986) Title: Acute toxicity ofCMATest Material Code 525 to Sheepshead Minnow, Cyprinodon variegatus This study is: ®

Adequate M

NotAdequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfurized, CASRN 68511-50-2 Remarks : Purity not provided METHOD Method/guideline followed: Test protocol followed OECDTG 203, Fish Acute Toxicity Test (1984) Test type: Fish acute toxicity test (WSF static renewal test) GLPcompliant?: Yes Year study was performed: 1986 Test species: Sheepshead minnow(Cyprinodon variegatus) Analytical monitoring: Yes; total organic carbon (TOC) measurements ofeach freshly prepared test solution and control and after 24-h on testjust before daily renewal with fresh test solution. Exposure duration: 96 hours Concentrations tested: 0or 10,000 mg/L (nominal loadingrates) Test method/conditions remarks: Test Organisms: source - a commercial supplier in New Hampshire; age - 24 to 29 days old ; total length - 20.3 nun average (range = 13 to 30 mm ; n = 18) ; wet weight - 0

.16 g average (range = 0 .03 to 0 .039 g; n =18) . Loading -0 .32 g biomass/L; pretreatment - none, fish held for a minimum of 20 days before testing. No feeding occurred during the test . Test System

: Individual WSFs were prepared for each daily renewal ofthe 10,000 mg/L test level . Ameasured weight oftest material was added to a measured volume ofdilution water (15-L) in a glass vessel and stirred for 16 to 24 hours. Stirring was accomplished using a Telfoncoated magnetic stir bar. Mixing speed was adjusted such that a vortex formed between 30 to 50%ofthe distance to the bottom . Following the mixing period, the test solutions were allowed to stand for 2 hours before the water phase was removed. To avoid removing test material fromFinal HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

the surface or bottom, a siphon was placed in the mixing vessel prior to addition of water and test substancewith the lower end approximately midway between bottomand surface. The siphoned water phase, designated water soluble fraction (WSF), was used for the aquatic toxicity test. About 90%

of the test solution in each test vessel was renewed daily after 24, 48, and 72 hours. Two 5-L replicates were used per treatment, and 10 fish were used per replicate (for a total of 20 per treatment) . Test vessels were loosely covered to reduce entry of dust, etc. Dilution Water : Natural seawater collected fromCape CodCanal, Bourne, Massachusetts which derives water fromBuzzards Bay or Massachusetts Bay . The water was filtered through 0 .5- micron polypropylene core filter and activated carbon, then stored for 1 to 4days prior to use while being constantly aerated. During storage, the water had a salinity of 32 to 33 ppt and pH of 7.7 to 7.8. During the test : dissolved oxygen - 5.6 mg/L to above 100%saturation (7.5 mg/L), pH - 6 .9 to 8 .0, salinity- 32, temperature - 20 to 22 °C . Mean measured TOClevels in the control and 10,000 mg/L WSF test level were 2.7 mg/L (range =1 .2 to 6 .0) and 4 .5 mg/L (range = 3.0 to 5.6), respectively. Test Levels: Control &10,000 mg/L WSF loading rate. Calculation ofLLsos: Statistical analysis of survival data not warranted. Test Substance: No undissolved test material was seen on the surface of the test vessels during the entire aquatic toxicity test . Reference Substance: Sodiumlauryl sulfate (SLS) . The 96-h LCso was 1 .2 mg/L. No information was provided on method of calculation . RESULTS Endpoint value(s) : Nominal concentrations : 96-h LLso > 10,000 mg/L; 96-h NOEC = 10,000 mg/L (no mortality or toxic signs noted) . Results remarks: No mortality or signs of toxicity were noted in the 10,000 mg/L WSFtest level and the control throughout the entire test. Mean measured TOCin the 10,000mg/L WSFtest level was 4 .5 mg/L compared to 2.7 mg/L in the control . Statistical results: Statistical analysis of survival data notwarranted. EPAdoes not consider the loading rate as the no-effect concentration when the concentration exceeds thewater solubility of the substance. Assuming that theexposure concentration in the WSF is the water solubility limit (saturation) for 1-propene, 2-methyl-, sulfurized, the no-effect concentration would be approximately 2 .7 mg/L. (No effects were observed at saturation.)Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

CONCLUSIONS No mortality or signs of toxicity were noted in the 10,000 mg/L WSF test level and the control throughout the entire test . STUDY RELIABILITY [2] Reliable with restrictions. The TOC in dilution water at the beginning and end ofthe test was greater than 2 mg/L rather than < 2 mg/L. REFERENCE Nicholson, R

.B. 1986. Acute toxicity of CMATest Material Code 525 to Sheepshead Minnow, Cyprinodon variegatus. Springbom Bionomics Study#10823-0186-6100-500-525, Report #BW-86-04-2004. As summarized in the Robust Summaries for theAlkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group ofthe American Chemistry Council's PetroleumAdditives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN_72162-15-3

C-2 . Acute Toxicity to Daphnia Summary of endpoint : Adequate Study 1 : Acute toxicity study ofCASRN68511-50-2 in Daphnia magna (Ward, 1993) Title: Acute Toxicity of the WaterAccommodated Fractions (WAFs) of CMA#613 to the Daphnid, Daphnia magna This study is: ®

Adequate 0

NotAdequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfurized, CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: U .S. EPA 797.1300 (1985, 1987), OECDTG 202 (1984) Test type: Daphnia sp., Acute Immobilization Test (Static) GLP compliant?: Yes Year study was performed: 1993 Test species: Water flea (Daphnia magna), less than 24-hours old, bred at lab facility Analytical monitoring: Yes; total organic carbon (TOC) measurements. Exposure duration: 48 hours Concentrations tested : 0, 100, 300, and 1000 mg/L WAF loading rates Test method/conditions remarks: A static non-renewal test was conducted using water accommodated fractions (WAF) ofthe test material at 100, 300, and 1000 mg/L loading rates. WAFs were prepared by adding a measured weight of the test material to a measured volume of the dilution water and stirring for 24 hours with a magnetic stir bar. The test solutions were allowed to stand for 1 hour before the water phase (WAF) was siphoned off . Test conditions: Two 250-ml, glass beakers that contained 200ml of test solution were used per treatment . The 250-mL test vessels were loosely covered to reduce entry ofdust, etc. Test temperature range: 20 t 1 °C

10Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Dilution water: Filtered well water collected at Hampton, New Hampshire and adjusted to the appropriate hardness 168 to 172 mg/L as CaC03 . The waterwas passed through activated carbon, a particle filter, and then an ultraviolet sterilizer and stored in a polyethylene tank where it was aerated. TOC levels were 2 mg/L at the beginning and end ofthe test, and 10 mg/L total suspended solids (TSS) at the beginning and < 10 mg/L at the end of the test . Lighting: A 16hour light and 8 hour dark photoperiod was maintained with cool-white fluorescent lights with an intensity of 201tEin'm 2 . Water chemistry: Dissolved oxygen - 8 .2 to 8 .7 mg/L; pH- 7.8 to 8

.2; conductivity - 570to 640ltmhos/cm

; temperature-20.5 to 20.9 °C. Element: Immobilization Test design: Control, 100, 300, &1000 mg/L WAF loading rates. 10 daphnids per replicate (20 per treatment) . Method of calculating mean measured concentrations: not applicable. Exposure period: 48 hours . Analytical monitoring: Total organic carbon (TOC) measurements ofinitial test solutions and control (0-hour) and at test termination (48-h) . TOClevels were 2 mg/L in the control, 3 mg/L at the 100 mg/L and 300 mg/L test levels; and 4to 5 mg/L at the 1000 mg/L test vessel . TOClevels were not considered to be indicative of actual test material concentrations and results are therefore based on nominal loading rates. Statistical methods were not utilized because there was greater than 50% survival in all test vessels. ° RESULTS Endpoint value(s) : 48-hour and 24-hour ECSo > 1000 mg/L (based on nominal loading rates) ; 48-hour and 24-hour NOEC = 1000 mg/L (based on nominal loading rates) . Results remarks: Effect concentrations were based on nominal loading rates. No immobilization was seen at the highest test concentration of 1000 mg/L (WAF) . Control response was satisfactory. EPAdoes notconsider the loading rate as the no-effect concentration when the concentration exceeds thewater solubility ofthe substance. Assuming that the exposure concentration in the WAF is the watersolubility limit (saturation) for 1-propene, 2-methyl-, sulfurized the no-effect concentration would be approximately 2.7 mg/L. (No effects were observed at saturation.) CONCLUSIONS The WAFs of the test material were not toxic to daphnids at the concentrations tested. Ninetyfive to 100%survival occurred at all test concentrations. No sublethal effects were noted during the test.Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

STUDY RELIABILITY [1] Reliable without restrictions. REFERENCE Ward, T .J . 1993. Acute Toxicity of the Water Accommodated Fractions (WAFs) of CMA#613 to the Daphnid, Daphnia magna. T .R. Wilbury Study#9178-CMA/ESI-613. As summarized in the Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPV Challenge Programby the Health, Environmental, and Regulatory Task Group ofthe American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

C-3 . Chronic Toxicity to Daphnia Summary of endpoint: Not adequate No studies were available for this endpoint .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

C-4 . Toxicity to Plants (Algae) Summary of endpoint: Adequate Study 1 : Toxicity ofCASRIV68511-50-2 to Pseudokirchneriella subcapitata Title: Not available This study is: ®

Adequate 0

Not Adequate TESTSUBSTANCE Identity: 1-Propene, 2-methyl-, sulfixrized, CASRN68511-50-2 Remarks : Purity not provided METHOD Method/guideline followed: Test protocol followed US EPAToxic Substances Control Act Test Guideline #797.1050 (1985, 1987), OECD TG 201 Alga, Growth Inhibition Test (1984) . Test type: Alga, Growth Inhibition Test (WAF static) GLP compliant?: Yes Year study was performed: 1994 Test species: Freshwater green algae (Pseudokirchneriella subcapitata) Analytical monitoring: Yes. Total organic carbon (TOC) measurements of initial test solutions andcontrol (0-hour) and at test termination (96-h) (EPA Method 415.1 (1979)) . Water samples were passed through a 0 .45 micron filter prior to TOC analysis. Exposure duration: 96 hours Concentrations tested: Control, 1 .0, 5.0, 10, 50, 100mg/L (nominal loading rates) Test method/conditions remarks: Test Organisms: source - T.R . Wilbury in-house culture originally purchased fromthe University ofTexas at Austin algae collection. Test System

: Individual WAFs were prepared for each test level and renewed daily . Ameasured weight of test material was added to a measured volume of dilution water (1-L) in a glass vessel and stirred for 24 hours. Stirring was accomplished using aTelfon-coated magnetic stir bar. Mixing speed was adjusted such that a vortex formed between 30 to 50% of the distance to the bottom . Following the mixing period, the test solutions were allowed to stand for 1 hour before the water phasewas removed. To avoid removing test material fromthe surface or bottom, a 14Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

siphon was placed in the mixing vessel prior to addition of water and test substance with the lower end 1-2 inches off the bottom . The siphoned water phase (i .e., WAF) was used for the aquatic toxicity test . A static test was conducted; i .e., there was no daily renewal oftest solution. Three 100-ml, replicates were used per treatment, with an inoculumof -10,000 cells/mL. The 250-mL Erlemneyer flasks were stoppered with foamplugs to reduce entry ofdust, etc. During the test, all treatment and control flasks were randomly placed on an orbital shaker adjusted to approximately 100cycles per minute under constant light (24 hours/day) . Dailycell counts were made visually by means of direct microscopic examination with a hemocytometer. Cool-white fluorescent lights provided a light intensity of47 to 50 WEin/m2sec. At the conclusion of the 96-h test, a 0 .5 mL subsample oftest media fromeach 100 mg/L test flask was combined with 100 mL of fresh untreated alga media and incubated for up to 9days or as soon as growth occurs. This was done to determine if growth inhibition was algistatic or algicidal . Dilution Water : Sterile enriched alga growth media (US EPA, 1978, T .R . Wilbury SOP#6) adjusted to pH 7.5. Measured TOC and total suspended solids in fresh untreated alga media were < 1 .0 and < 10 mg/L, respectively. Test temperature - 23.4 to 23.6 C ; pH- 7.0 to 7.1 at 0- hour and 8.6 to 10.2 after 96 hours. TOCmeasurements were only made on the lowest and highest test levels and control at the beginning and end ofthe test . TOC levels were < 1 .0 mg/L in the control and 1 .0 mg/L WAF test level and 3 mg/L at 100 mg/L. Test Levels: Control, 1 .0, 5.0, 10, 50, 100 mg/L WAF loading rates. Calculation of ELsos and NOELRs: Moving average and probit methods (Stephan, 1983) were used to calculate ECsos (i .e., ELscs) . Aparametric one-wayanalysis of variance (ANOVA) and Dunnett's test were used to calculate the no-observed effect concentration (i .e., EL.os) when data were normally distributed and a non-parametric Kruskal and Wallis test was used if data were not normally distributed. Test Substance: No undissolved test material was seen on the surface ofthetest vessels during the entire aquatic toxicity test . Reference Substance: No RESULTS Endpoint value(s) : Based on nominal loading rates: 72-hour EL50 (cell density) = 26 (21-32) mg/L; 72-hour NOELR(cell density) = 5.0 mg/L; 96-hour EL50 (cell density) = 34 (29-39) mg/L; 96-hour NOELR(cell density) =10 mg/L; 72-hour EL50 (growth rate) > 100 mg/L; 72- hour NOELR (growth rate) = 5.0 mg/L; 96-hour ELso (growth rate) > 100 mg/L; 96-hour NOELR (growth rate) = 10 mg/L Results remarks: Test concentrations for the definitive test were not specified in a protocol amendment and the pH ofthe sterile media at the start ofthe test was 7.0 rather than 7.5. These deviations did not compromise the study . 15Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

72-hour ELso for cell density was determined by moving average method. 96-hour ELSo for cell density was determined by Probit method. 96-hour NOELRs for cell density and growth rate were determined by hypothesis analysis tests. Re-growth of inhibited cultures fromthe 100mg/L test level revealed the effect was algistatic rather than algicidal . CONCLUSIONS After 96 hours of exposure, reductions in cell density and growth rate were observed at a loading rate > 10 mg/L. STUDY RELIABILITY [I] Reliable without restrictions. REFERENCE Reference information is not available. This studywas obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Programby the Health, Environmental, and Regulatory Task Group ofthe American Chemistry Council's Petroleum

AdditivesTanel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

C-5 . Acute Toxicity Summary of endpoint : Adequate Study 1 : Acute oral toxicity study ofCASRN685II-50-2 in rats Title: Not available This study is: ®

Adequate 0

Not Adequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfiu-ized, CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: Not specified Test type: Acute Oral Toxicity (LD5o) GLPcompliant?: Pre-GLP Year study performed: 1975 Test species : Sherman/Wistar rat (young adult) Number of animals/sex/dose: 5 males/dose Route of administration: Oral gavage Vehicle: None Doses/concentrations tested: 2 .0, 4 .0, 8 .0, 16.0, or 32.0 mL/kg Statistical methods used: Not specified Test method/conditions remarks: Rats were fasted 24 hours prior to dosing. Test material was administered by gavage in a single oral dose at concentrations of 2.0, 4 .0, 8.0, 16.0, or 32.0 mL/kg. Animals observed for 14 days post-dosing for signs oftoxicity or mortality . Body weights were not taken

; gross necropsies and histopathology were not performed.Final HPV3 Test Plan and Data Adequacv Review CASRN 72162-15-3

RESULTS Endpoint value(s) : LDso = 5.7 mL/kg ; 19/20confidence limits = 4-8 mL/kg Results remarks: No deaths were observed at 2.0 or 4 .0 mL/kg. At 8 mL/kg, 4/5 were dead at day 1 post dosing and 1/5 were dead at day 2 . 5/5 rats were dead at day 1 in groups given 16.0 or 32.0 mL/kg. No data were presented on parameters other than mortality. CONCLUSIONS The oral LDSo is 5.7 mL/kg (-5700 mg/kg) for male rats. STUDY RELIABILITY [2] Reliable with restrictions. Details of study design andobservations during study were not presented. Body weights were not measured; gross necropsies and histopathology were not performed. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 2: Acute oral toxicity study of CASRIV68515-88-8 in rats 

### Title: Not available This study is: ®

Adequate 0

Not Adequate TEST SUBSTANCE Identity: Pentene, 2,4,4,-trimethyl, sulfurized, CASRN68515-88-8 Remarks : Purity not provided METHOD Method/guideline followed: Consistent with guidelines outlined in OECDTG 401 Test type: Acute Oral Toxicity GLP compliant?: Yes Year study performed: 1987 Test species: Albino rats of the outbred Sprague-Dawley strain Number of animals/sex/dose : 5/sex/dose Route of administration: Oral gavage Vehicle: Mineral oil-based material Doses/concentrations tested: 5.0 g/kg Statistical methods used: Not specified Test method/conditions remarks: The animals were fasted overnight before dosing. The sample was administered as supplied at a limit dose of5.0 g/kg via oral gavage with a syringe and Nelaton catheter . Following administration, the animals were allowed food andwater for the 15-day observation period. The animals were observed three times on the day ofdosing and twice on study day two and daily thereafter. Individual weights were recorded on the day of dosage and on the day of termination (day 15) . The animals were euthanized by carbon dioxide at the conclusion ofthe observation period. Gross autopsies were performed on all animals that died during the observation period and on all survivors after day 15.Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

RESULTS Endpoint value(s) : LDSo > 5.0 g/kg Results remarks: All animals survived to termination ofthe experiment (day 15) . Decreased activity (3/5 females), diarrhea (1/5 males), salivation (1/5 males and 1/5 females), and apparent urinary incontinence (3/5 females) were noted following dose administration. All animals appeared normal on study days 5-15. Test material did not cause an adverse effect on mean body weight in either sex. CONCLUSIONS The oral LDSo is greater than 5.0 g/kg for male and female rats. STUDY RELIABILITY [1] Reliable without restrictions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Planand Data Adeouacv Review CASRN72162-15-3

Study 3 : Acute dermal toxicity study ofCASRIV68515-88-8 in rabbits Title : Not available This study is: ®

Adequate F~ Not Adequate TEST SUBSTANCE Identity : Pentene, 2,4,4-trimethyl, sulfiuized, CASRN 68515-88-8 Remarks: 97%

purity METHOD Method/guideline followed: FHSA Regulations 16 CFR 1500.40 Test type: Acute Dermal Toxicity GLPcompliant?: Yes Year study performed: 1986 Test species: New

Zealand White rabbits (young adults) Number of animals/sex/dose: 5/sex/dose Route of administration: Dermal Vehicle: Mineral oil-based material dosed undiluted Doses/concentrations tested: 2.0 g/kg Statistical methods used: Not specified Test method/conditions remarks: The sample was applied to unabraded shaved skin under impervious occlusion for 24 hours at a limit dose of 2 .0 g/kg. At the end of the 24-hour exposure period, the wrapping was removed and any unabsorbed test material remaining on the skin was removed by gentle sponging using a paper towel moistened with mineral oil . The animals were observed for signs of toxicity or behavioral changes frequently on the day oftreatment . Thereafter, all surviving rabbits were examined for outward signs oftoxicity once per day, for the entire 14-day observation period. Individual weights were recorded on the dayof dosage, weekly thereafter and prior to sacrifice. The surviving animals were euthanized at the conclusion of the observation period. Gross autopsies were performed on all animals after 14 days.Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

RESULTS Endpoint value(s) : LDSO > 2.0 g/kg Results remarks: One rabbit died on day 14 ofthe study. No other deaths were observed during the 14-day observation period. In the male group, mild skin erythema and mild-to-moderate edema were observed after unwrapping at 24 hours. Slight to mild skin irritation noted at day 7 was completely resolved by day 14. Aloss of body weight was noted for 1/5 male animals at day 7. The same animal was found dead on day 14 after experiencing a bloated appearance. Signs of dehydration and no formed fecal material in the intestinal tract were noted for the one mortality. Other than the previous observation, all animals appeared normal throughout the 14- dayobservation period. In the females, skin reactions were typical ofthose observed with the male group. A loss ofbody weight was noted in one female at day 7. Gross pathological examination of the female rabbits revealed no remarkable findings. CONCLUSIONS Thetest article, when dosed as supplied and studied in 5 males and 5 female albino rabbits, appears to have an acute dermal LDSo greater than 2.0 g/kg. STUDY RELIABILITY [1] Reliable without restrictions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Programby the Health, Environmental, and Regulatory Task Group ofthe American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adeauacv Review CASRN 72162-15- 3

Study 4 : Acute dermal toxicity study ofCASRN67762-55-4 in rabbits Title: Notavailable This study is : ®

Adequate F Not Adequate TESTSUBSTANCE Identity: Alkenes, C15-18 alpha-, sulfurized, CASRN 67762-55-4 Remarks: Purity not provided METHOD Method/guideline followed: Not indicated Test type: Acute Dermal Toxicity GLP compliant?: Yes Year study performed: 1996 Test species: New

Zealand White rabbits Number of animals/sex/dose: 10/sex/dose Route of administration: Dermal Vehicle: None Doses/concentrations tested: 2000 mg/kg Statistical methods used: Not specified Test method/conditions remarks: One dermal, semi-occluded patch of test article at 2000 mg/kg was applied to clipped dorsal skin of each animal . The patches were removed after 24 hours. All animals were observed daily for 14 days following test article administration. RESULTS Endpoint value(s) : LDSp > 2000 mg/kg Results remarks: No clinical signs were observed during the study. Erythema and/or edema of the skin at application site were observed on Day 1 in some animals. Therewas an increase in mean body weight during the study . None ofthe animals died during the study. No visible lesions were observed in any animal at terminal necropsy. 23Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

CONCLUSIONS The dermal LD5o is greater than 2000mg/kg . STUDY RELIABILITY [1] Reliable without restrictions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacv Review __ CASRN 72162-15-3

Study 5: Acute inhalation toxicity study ofCASRIV685II-50-2 in rats Title: Not available This study is: ®

Adequate 0

Not Adequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfurized, CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: Modified OECDTG 403 

### Test type: Acute Inhalation Toxicity GLPcompliant? : Yes Year study performed: 1990 Test species: Sprague-Dawley rats (9 weeks old at initiation of study) Number of animals/sex/dose: 10/sex/dose Route of administration : Whole-body inhalation of vapor Vehicle: Not applicable Doses/concentrations tested: 0 .07 or 0 .39 mg/L Statistical methods used: Not specified Test method/conditions remarks : Rats were exposed to 0 .07 or 0 .39mg/L vapor for a single 4-hour whole-body exposure. Shamcontrol rats were placed in inhalation chambers in room air. One half ofrats (5M, 5F) fromeach group were sacrificed 24 hour post-exposure; others maintained for 2 weeks recovery. Vapors fromthe methyl propene derivative were generated fromsample heated to 200°C in a counter current generator and delivered into the exposure chamber. Air samples were pulled through glass fiber filters to verify that particles were not present and animals were being exposed to pure vapor. Vapors were analyzed by GC to quantitate chamber concentrations using octane as a standard; qualitative analyses were performed by GC/MS. Animals were observed in the chambers and post exposure for clinical signs of toxicity. Body weights were taken and selected organs weighedat necropsy. Histopathology was conducted on liver, kidney, lungs, nasal turbinates, and tracheobronchial lymph nodes. 25Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

RESULTS Endpoint value(s) : LCso > 0 .39 mg/L Results remarks: No mortality and minimal toxicity were observed. Abnormal clinical signs occurring during and immediately post-exposure included oral and ocular discharge, shallow respiration (some high-dose rats only), and decreased response to stimuli . No abnormal treatment-related clinical signs were observed from1 hour post-exposure to the end of the recovery period. Group body weights and weights ofliver, kidney, and lungs were unaffected by exposure. No treatment-related microscopic lesions were observed in the 5 organs examined. CONCLUSIONS The LCSO > 0.39 mg/L was for avapor generated fromthe starting material at 200°C, not from the whole methyl propene derivative. Vapor approximates conditions ofworkplace exposure. STUDY RELIABILITY [2] Reliable with restrictions. Study performed with only two dose groups and is not a limit test . Physical and chemical properties provided with exception of structural formula and purity of starting material . REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Programby the Health, Environmental, and Regulatory Task Group ofthe American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 6 : Acute inhalation toxicity study ofCASRN685I5-88-8 in mice andguinea pigs Title: Not available This study is: ®

Adequate 0

Not Adequate TEST SUBSTANCE Identity: Pentene, 2,4,4-trimethyl-, sulfurized, CASRN68515-88-8 Remarks: Purity not provided METHOD Method/guideline followed: Consistent with EPAHealth Effects Guideline OPPTS 870 .1300 Test type: Acute Inhalation Toxicity GLPcompliant?: Yes Year study performed: 1988 Test species : CD-1 strain mice, Hartley strain guinea pigs Number of animals/sex/dose: 5/sex/dose Route of administration: Aerosol inhalation (whole-body) Vehicle: Mineral oil-based material, dosed as supplied Doses/concentrations tested: 4 .3 mg/L Statistical methods used: Not specified Test method/conditions remarks: Two groups of five mice/sex and five guinea pigs/sex were exposed for 4hours to the test material (nominal concentration of 5 mg/L) as a liquid droplet aerosol generated by a Laskin nebulizer apparatus delivered into a plexi-glass chamber. Also, a control group of mice and guinea pigs was exposed to mineral oil in the same manner as the testmaterial-exposed group except that the test material was not administrated. The details of the whole-body exposure are consistent with those described in EPA Health Effects Guideline OPPTS 870

.1300 . The actual exposure concentration as measured by gravimetric analysis was 4 .3 mg/L. Particle size analyses were performed once/hour fromthetest material chamber using a cascade impactor. Animal observations for toxicological signs and mortality were recorded every 15 minutes during the exposure, and twice daily for the 14-day observation period. Individual weights were recorded on the day prior to exposure and on days 2, 3, 5, 8and 14. At theconclusion ofthe observation period, the surviving animals were euthanized by

27Final HPV3 Test Plan and Data Adeauacv Review CASRN72162-15-3

exsanguination under general anesthesia. All animals were subjected to gross necropsy (nasal passages, trachea, external surface, all orifices, the cranial cavity, the brain and spinal cord, and all viscera) . RESULTS Endpoint value(s) : LCSO (mice) > 4 .3 mg/L; LCso (guinea pigs) > 4 .3 mg/L Results remarks: The mass median aerodynamic diameter for the studies was 1 .6 microns with a geometric standard deviation of 2.1 (estimated percent of particles < 10 microns = 100%) . One female guinea pig was euthanized on study day 7 because of a broken leg, an effect thought to be unrelated to the administration to the test material . All other animals survived the duration ofthe study. Observations noted during the test material exposure included reduced activity and matted coat. Signs exhibited by the test animals upon removal fromthe chamber and during the two-hour post-exposure observation period on day 1 included matted coat, yellow fur, yellow ano-gentital staining, and nasal discharge. The control groups were generally unremarkable during the exposure and immediately thereafter. During week 1, thetest mice exhibited few signs other than matted coat . The test guinea pigs exhibited matted coat and ano-genital staining. During week 2, ano-genital staining was theonly remarkable sign in the guinea pigs. No significant difference was noted between the test and control group weights for either species. No gross lesions that could be attributable to the test material were observed in any ofthe mice or guinea pigs. CONCLUSIONS Ten often CD-1 mice and ten of ten Hartley guinea pigs received a single four-hour whole-body exposure to 4

.3 mg/L test material as a respirable aerosol . All animals survived the exposure and the 14-day post-exposure observation period with the exception of a single guinea pig that was euthanized for a broken limb. Signs oftreatment included reduced activity and matted coat during the exposure. Thetreated animals were generally comparable to air-only control animals during the observation period. Body weight values and gross post mortem observations were generally unremarkable for differences between control and treated animals of either species. STUDY RELIABILITY [1] Reliable without restrictions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This studywas obtained from the Robust Summaries for theAlkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

Study 7: Acute inhalation toxicity study ofCASRN68515-88-8in mice, rats, andguinea pigs Title: Not available This study is: ®

Adequate 7 Not Adequate TEST SUBSTANCE Identity: Pentene, 2,4,4-trimethyl-, sulfurized, CASRN68515-88-8 Remarks: Purity not provided METHOD Method/guideline followed: Consistent with EPA Health Effects Guideline OPPTS 870 .1300 Test type: Acute Inhalation Toxicity GLPcompliant?: Yes Year study performed: 1988 Test species : CD-1 COBS Swiss Albino mice, Sprague-Dawley CDrats, Hartley guinea pigs Number of animals/sex/dose: 5/sex/dose Route of administration: Aerosol inhalation (whole-body) Vehicle: Mineral oil-based material, dosed as supplied Doses/concentrations tested: 4 .3 mg/L Statistical methods used: Not specified Test method/conditions remarks: Group of five mice/sex, five rats/sex, and five guinea pigs/sex were exposed for 4 hours to the test material as a liquid droplet aerosol generated by a Laskin nebulizer apparatus delivered into a plexi-glass chamber. Also, control groups of mice, rats and guinea pigs were exposed to mineral oil in the same manner as the test-material-exposed group except that the test material was not administrated. The details ofthe whole-body exposure are consistent with those described in EPAHealth Effects Guideline OPPTS 870 .1300 . The actual exposure concentration as measured by gravimetric analysis was 4 .3 mg/L. Particle size analyses were performed once/hour fromthe test material chamber using a cascade impactor . Animal observations for toxicological signs and mortality were recorded every 15 minutes during the exposure, and twice daily for the 14-day observation period. Individual weights were recorded on the day prior to exposure and on days 2, 3, 5, 8, and 14. At the conclusion of the observation period, the surviving animals were euthanized by exsanguination 29Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

under general anesthesia. All animals were subjected to gross necropsy (nasal passages, trachea, external surface, all orifices, the cranial cavity, the brain and spinal cord, and all viscera) . RESULTS Endpoint value(s) : LC50 (mice) > 4 .3 mg/L; LCso (rat) < 4

.3 mg/L; LCso (guinea pigs) > 4 .3 mg/L Results remarks : The test animals received an average analytical exposure concentration of 4 .3 mg/L test material with a nominal exposure concentration of 100 mg/L. Particle size distribution measurements showed an average mass median aerodynamic diameter of 3.8 microns with an average geometric standard deviation of 1 .9 microns. Approximately 93 percent ofthe aerosol was 10 microns or less in size. Mortality and physical observations : Three female rats died within a day after exposure. A single male mouse and a single male guinea pig also died on test days 7 and 9, respectively. All other animals survived the duration ofthe study. Observations noted during exposure included nasal discharge, salivation, closed eyes and wet fur. Signs exhibited by the rats upon removal fromthe chamber and during the two-hour post-exposure observation period on day included numerous secretory responses, labored breathing, rales andwet fur. Also, several ofthe females showed tremors. One ofthe female rats died two hours after exposure. The mice and the guinea pigs were generally unremarkable except for contaminated fur. Two additional rats were found dead the morning after exposure. The surviving rats (both sexes) continued to show responses without a complete recovery during the 14-day post-exposure observation period, including nasal discharge, labored breathing, rales, andcontaminated fur leading to alopecia. Body weight : Significant body weight losses were observed following exposure amongall three species. The surviving mice and rats began to recoverweight within a week after exposure. However, guinea pigs continued to lose weight throughout the first week and did not showa weight gain until the end of the second week. Gross post mortemobservations: Discoloration ofthe lungs and nasal turbinates was noted among the spontaneously dying animals. CONCLUSIONS The LC50values are as follows: LC50 (mice) > 4 .3 mg/L; LCso (rat) < 4

.3 mg/L; LCso (guinea pig) > 4 .3 mg/L. STUDY RELIABILITY [2] Reliable with restrictions. Only one concentration was tested and mortality was observed in rats; a full study should have been conducted to determine the LC50 for rats.Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3



### REFERENCE Reference information is not available (the underlying study contains confidential business information) . This studywas obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPV Challenge Program by the Health, Environmental, and Regulatory Task Group ofthe American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

Study 8 : Acute inhalation toxicity study ofCASRN685I5-88-8in rats Title : Not available This study is : ®

Adequate 0Not Adequate TEST SUBSTANCE Identity: Pentene, 2,4,4-trimethyl-, sulfurized, CASRN 68515-88-8 Remarks: Purity not provided METHOD Method/guideline followed: OECDTG 403 Test type: Acute Inhalation Toxicity GLPcompliant?: Yes Year study performed: 1987 Test species : Sprague-Dawley Albino rats Number of animals/sex/dose: 5/sex/dose Route of administration: Aerosol inhalation (whole-body) Vehicle: Mineral oil-based material dosed undiluted Doses/concentrations tested: 1 .5, 2 .5, and 5.6 mg/L (measured) Statistical methods used : The LCso with 95%confidence intervals was computed using the method ofMiller and Tainter (1944) . Test method/conditions remarks: Three groups offive rats/sex were exposed for 4 hours to the test material as a liquid droplet aerosol generated by a pressure spray apparatus delivered into a plexi-glass chamber. The details ofthe whole-body exposure are consistent with those described in OECDTG 403 . The actual exposure concentrations as measured by gravimetric analysis were 1 .5, 2

.5, and 5.6 mg/L. Particle size analyses were performed twice/hour using a multi-stage cascade impactor . Animal observations for toxicological signs and mortality were recorded periodically during the exposure, and twice daily for the 14-day observation period. Individual weights were recorded on the day prior to exposure and on days 4, 8, and 14. At the conclusion ofthe observation period, the surviving animals were euthanized using pentobarbital as an anesthetic followed by exsanguination. All animals were subjected to gross necropsy (external body surface and orifices, major visceral organs, body cavities and carcass) . 32Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3



### RESULTS Endpoint value(s) : LCso (males) > 5.6 mg/L; LCso (females) = 2.17 mg/L Results remarks: The mass median aerodynamic diameter for the studies was 3.15 microns with a geometric standard deviation of 2.45 (estimated percent ofparticles < 12 microns = 90.5%)

. Remarkable animal observations during the studies include alopecia (noted at all dose levels during second week of observation), ataxia (noted prior to thedeath ofone female in the 5.6 mg/L group), dark material around eye (noted in two animals/sex at the 5.6 mg/L dose), decreased activity (noted in all animals at the dose level of 2.5 and 5.6 mg/L; reversible by study day 5), respiratory irregularity (increased respiration noted in all groups during and immediately following exposure; reversible by study day 7), tremors (noted in one female during and immediately following exposure to 2.5 mg/L) . No male deaths were recorded for any ofthe dose levels. Group mean body weights were decreased at day 4 among males exposed to 2.5 and 5.6 mg/L. This effect was reversible by study observationday 8and 14. Three of 5 females in the 1 .5 mg/L group died on day 2 following exposure. Four of 5 female rats exposed to 2.5 mg/L died on observation day 2 . Three females in the high-dose group died on day 2 following exposure, with an additional death on day 6 . Body weights decreased at day 4 in the surviving females, an effect that was reversible by days 8and 14. No internal lesions or abnormalities were noted in any animal sacrificed at study termination. Pathological findings among females which died during the course ofthe observation period were observed in several organs, including the brain (prominent vascularization, and blood in the cranial cavity), nasal passages (reddening of the nasal passage, with the notation ofclear fluid in the nasal passage), lungs (reddening of the lungs, with the observation of a `puffy' lung in one female), and trachea (clear fluid noted in the trachea of one female) . CONCLUSIONS Following 4-hour whole-body exposure to a liquid droplet aerosol of the test material, the LC5o in male Sprague-Dawley rats is considered to be greater than 5.6 mg/L. The LCso value in females was calculated to be 2.17 mg/L with upper and lower confidence limits of 3.69 and 0 .64 mg/L, respectively. STUDY RELIABILITY [1] Reliable without restrictions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPA under the HPV Challenge Programby the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's PetroleumAdditives Panel on May 19, 2005) .

33Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

C-6 . Repeated-Dose Toxicity Summary of endpoint : Adequate Study 1 : Repeated-dose dermal toxicity study ofCASRN68511-50-2 in rats Title : Not available This study is : ®

Adequate 0Not Adequate TEST SUBSTANCE Identity : 1-Propene, 2-methyl-, sulfiuized CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: Comparable to OPPTS 870 .3250 Test type: Thirteen-week dermal subchronic toxicity study GLPcompliant?: Yes Year study performed: 1989 Test species: Sprague-Dawley (Tac:N[SD)fBR) Number of animals/sex/dose: 10/sex/dose Route of administration: Dermal Vehicle: Mineral oil (solvent-refined naphthenic base stock), density 0 .88 g/mL Exposure duration: 5 days/week, 13 weeks Doses/concentrations tested: Part 1 : 500and 2000 mg/kg/day undiluted test material ; 500 mg/kg/day diluted 50% w/v in mineral oil base stock; Part 2 : 500, 250, 100, 50, and 10 mg/kg/day diluted in mineral oil base stock Statistical methods used : Analysis of Variance followed by multiple range tests; serum chemistry and hematology data were evaluated using the F test forANOVAand StudentNewman-Keuls multiple comparison test . Test method/conditions remarks: Age at initiation: 7 weeks old following 2weeks acclimation . 34Final HPV3 Test Plan and Data Adeauacy Review CASRN 72162-15-3

Studywas designed to identify inherent toxicity oftest material and to determine whether dilution in a mineral oil carrier would alter toxicity. Methyl propenederivative was applied to the clipped backs of groups of 20 Sprague-Dawley rats (1 OM, l OF) 5 days per week for 13 weeks at dose levels of 500or 2000 mg/kg/day undiluted or at dose levels of 500, 250, 100, 50, or 10 mg/kg/day diluted in mineral oil on the same schedule. Rats were fitted with Elizabethan collars to minimize ingestion oftest material, which was left uncovered on the skin. One vehicle and 2 untreated shaved control groups were included in the study . Assessments for toxic response included: daily clinical observations ; weekly skin irritation scoring ; weekly body weights; terminal organ weights; hematology, serum chemistry, and urinalysis at weeks 5 and 13; and gross necropsy evaluations, sperm morphology, and histopathology at study termination . Endpoint value(s) : LOAEL = 100mg/kg/day based on increased production of white blood cells in spleen and bone marrow ; NOAEL = 50 mg/kg/day Results remarks : Male rats treatedwith methyl propene derivative for 13 weeks at dose levels >_ 250mg/kg/day gained less weight (15%less at study termination) than controls. Female weights were unaffected. At doses ? 250mg/kg/day, both sexes had decreased levels of red blood cells and increased levels ofneutrophils in circulation, increased spleen size and increased pigment and red pulp in the spleen. At doses >_ 100mg/kg/day, there was increased production ofWBCin spleen and bone marrow . Mean liver-to-body weights were increased in male rats at dose levels >_ 250mg/kg and in female rats at >_ 500 mg/kg/day. Male rats treated with undiluted test material at 500or 2000mg/kg/day had increased kidney weights correlated with dose-related increase in hyalinedroplet formation indicative of light hydrocarbon nephropathy. Undiluted test material and dilutions at 25% (500 mg/kg, 250mg/kg in Part 2) induced moderate to strong reaction in the skin, characterized by erythema, edema, increased thickness, and stiffness; these effects were more severe in the 500mg/kg (diluted 50% w/v) group. Microscopically, hyperkeratosis, hyperplasia of sebaceous gland, increased mitosis in epidermis, and dermal abscesses were observed. Virtually no irritation was observed in the vehicle control group or in dose groups of 100, 50, or 10 mg/kg/day where dilutions were made at 10% or 5% w/v . No effects on sperm

motility or morphology were observed in rats treated with 2000 mg/kg/day. The relative weight increases in livers of higher dose animals of both sexes had no microscopic correlate and is considered an adaptive response to treatment. The increase in kidney weight and hyaline droplet formation in male rats is indicative of light hydrocarbon nephropathy, a condition considered by EPAto be specific to male rats and not predictive ofcomparable toxicity in humans . Although many changes in hematology parameters can be associated with infections which can occur with severe skin irritation, increased dose-related neutrophil production at >_ 250 mg/kg/day was observed in animals with minimal skin irritation and can be considered adirect effect ofthe test material . CONCLUSIONS The NOAEL for systemic toxicity is 50mg/kg/day basedon increased production ofwhite blood cells in spleen and bone marrow at 100mg/kg/day. The minimally irritating concentration of methyl propene derivative diluted in mineral oil base stock is 10% . 35Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

STUDY RELIABILITY [1] Reliable without restrictions. REFERENCES Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 2: Repeated-dose dermal toxicity study of CASRN685II-50-2 in rabbits (28 days) Title: Not available This study is: ®

Adequate F NotAdequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfiuized, CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: Reference for study design: Federal Register, Volume 43, Number 163 Test type: 28-Day Subchronic Dermal Toxicity Study GLP compliant?: Yes Year study performed: 1982 Test species: Albino rabbits Number of animals/sex/dose: 6/sex/dose Route of administration: Dermal Vehicle: None Exposure duration: 4weeks Doses/concentrations tested: Group 1 : 200mg/kg/day undiluted test material ; Group 2 : 2000 mg/kg/day undiluted test material Statistical methods used: All data was submitted to analysis ofvariance (method: Statistical Analysis System, SAS Statistical Institute 1979) followed by evaluation using the NewmanKeuls method for all significant dose differences. Test method/conditions remarks: Age at initiation oftreatment: Not specified. Treatment beganfollowing 1 week acclimation. Number of animals tested: 36 animals (6M, 6F/group) : 2 treatment groups, 1 untreated control group. Study was designed to evaluate the subchronic toxicity ofthe test material when applied dermally. Methyl propene derivative was applied to the shaved dorsal trunk area (approximately 10%of the body surface) of 2 groups of 12 albino rabbits (6M, 6F) 5 days perweek for 4weeks at dose levels of200or 2000 mg/kg/dayof 37Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

undiluted test material on the same test schedule. Halfof the animals in each group were abraded once perweek throughout the study . The abrasion penetrated the stratumcorneum, but did not cause bleeding. The treated skin was occluded for at least 6hours daily and the trunk of each animal covered with an impervious material . One untreated shaved control group of 6 animals (3 intact, 3 abraded) was included in the study. Assessments for local and systemic effects included: clinical observations; skin irritation scoring 5 days per week; body weights (at pretest, every 3 to 4days during testing, termination) ; hematology, serum chemistry, and urinalysis at pretest andtermination ; and gross necropsy evaluations at study termination. Endpoint value(s) : LOAEL = 2000 mg/kg/day based on weight loss in males and increased monocytes; NOAEL = 200mg/kg/day Results remarks: One male rabbit death occurred at the higher dose level . Body weight gains were observed in control (0.5 to 1 .0 kg) and lower dose group (0.2 to 1 .0 kg) . A trend ofweight loss and change in food consumption among the high-dose males was observed in the latter half ofthe study

. Weight gain was observed in 5 of 6 high-dose females. Treatment with the test material caused severe skin irritation at both doses. Abrasion of skin increased the degree of irritation at the low-dose level . No irritation was observed in the control group. Urinalysis values were normal for all groups. The low-dose group showed an increase in chloride and a decrease in albumin

. The high-dose group showed decreased alkaline phosphatase and an increase in chloride and globulin. Hematology showed no trends in the control and low-dose groups while monocyte determinations were significantly different (increased) in the high-dose group . Gross and histopathological examination of tissues did not reveal any pattern of changes attributable to dermal contact with the test material . At autopsy, one animal in the control group was found to be female instead of male and one animal in the low-dose group was found to be male instead of female. Statistical evaluation including and excluding these two animals showed no significant differences. CONCLUSIONS LOAEL = 2000 mg/kg/day ; NOAEL = 200mg/kg/day. No minimally irritating concentration was identified by this study. STUDY RELIABILITY [2] Reliable with restrictions. Animal ages were not included in the report . An uneven sex distribution was used. Clinical behavior determinations beyond that of morbidity were not included in the report . Only two doses were tested. REFERENCES Reference information is not available (the underlying study contains confidential business information) . This study was obtained from the Robust Summaries for theAlkyl Sulfides Category (submitted to EPAunder the HPV Challenge Programby the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) . 38Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

Study 3: Repeated-dose dermal toxicity study ofCASRN685II-50-2 in rabbits (2l days) Title: Not available This study is: ®

Adequate RNot Adequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfurized, CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: None cited Test type: 21-Day Repeated Dose Dermal Toxicity Study GLPcompliant?: No Year study performed: 1979 Test species : New

Zealand White rabbit Number of animals/sex/dose: 3 treatment groups (7M/3F, 8M/2F, 6M/4F), 1 untreated control (5M/5F) Route of administration : Dermal Vehicle: None Exposure duration: 21 days Doses/concentrations tested: 140, 560, and 2240 mg/kg/day undiluted test material Statistical methods used: Not available Test method/conditions remarks: Age of animals at initiation: Not specified. Exposure was initiated following at least 2 weeks acclimation . Study was designed to evaluate local and systemic effects of test material when applied dermally. Methyl propene derivative was applied to the shaved backs and sides (approximately 10%of the body surface) of 3 groups of 10 New ZealandWhite rabbits 5 days per week for 3 weeks at dose levels of 140, 560, or 2240 mg/kg/day ofundiluted test material on the same test schedule. The animals were fitted with plastic collars to inhibit ingestion of the test material, which was left uncovered on the skin and not removed prior to the next dose. One untreated shaved control group of 10 animals was included in the study

. Assessments for local and systemic effects included: twice daily (morning 39Final HPV3 Test Plan and Data Adequacv Review CASRN 72162-15-3

and afternoon) clinical observations; skin irritation scoring 5 days per week; weekly body weights; hematology, serum chemistry, and urinalysis at pretest and termination ; and gross necropsy evaluations at study termination. Endpoint value(s) : NOAEL = 2240 mg/kg/day basedon no systemic effects at the highest dose tested Results remarks: All rabbits survived the duration of the test . Body weight changes were within expected ranges and comparable for all groups. Rabbits in all groups had lethargy, ptosis, G .I . disturbances, nasal and ocular discharges, and respiratory distress, all more often in the second and thirdweeks with no discernible pattern ofresponse. Skin responses included slight to moderate erythema and very slight to slight edema during Week 1 for all treated groups. During Week 2, responses in all treated groups were moderate to severe erythema with additional signs ofcracked skin, bleeding, and discoloration. Edema was slight at the lowest dose and slight to moderate at the higher doses. During Week 3, all treated groups had severe erythema with cracked and bleeding skin, eschar, and discoloration and edemawas slight to moderate. No irritation was observed in the control group. Urinalysis values were normal in all groups. Several individual and isolated hematological and serumchemistry values were out of expected range, but with no discernible treatment related changes in the mean values for all groups. At necropsy, sporadic occurrences of dark lungs and liver, red and bloated intestines, pale kidney, or small or gray spleen were noted with no relationship to treatment . Epithelial hyperplasia of the treated skin was observed in all rabbits with the treated groups exhibiting slightly more severe grades of hyperplasia than the control group. The rabbits were grouped by sex at the start of the study. At necropsy, six errors in sexing were discovered which resulted in uneven sex distribution within the groups. Since there were no apparent effect differences between the sexes, the study is not considered to be compromised. With no discernable pattern of response in both test andcontrol groups, observed clinical signs are considered to be related to handling. The occurrence of hyperplasia in all groups suggests a relationship to clipping rather to test material administration. However, in-life dermal observations revealed severe erythema responses in all treated rabbits and none in the sham-treated control group. CONCLUSIONS The NOAEL is 2240 mg/kg/day based on no systemic effects at the highest dose tested. No minimally irritating concentration was identified by this study . STUDY RELIABILITY [2] Reliable with restrictions. Animal age and organ weight data were not included in the report . The test site was not occluded following test material application . An uneven sex ratio was used. REFERENCES Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, 40Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

and Regulatory Task Group of the American Chemistry Council's PetroleumAdditives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 4 : Repeated-dose inhalation toxicity study ofCASRN68515-88-8 in rats Title: Not available This study is: ®

Adequate F] Not Adequate TEST SUBSTANCE Identity: Pentene, 2,4,4-trimethyl-, sulfurized, CASRN68515-88-8 Remarks : Purity not provided METHOD Method/guideline followed : OECDTG 412 Test type: 4-Week Inhalation Toxicity Study in Rats GLPcompliant?: Yes Year study performed: 1989 Test species : Sprague-Dawley CDrats (7 weeks old at initiation of treatment) Number of animals/sex/dose: 10/sex/dose for low- and mid-dose levels; 15 male and 20female rats for the control and high-dose level Route of administration: Aerosol inhalation (whole-body) Vehicle: None 

### Exposure duration: 4 weeks followed by a 3-week recovery period Doses/concentrations tested: 0, 15, 50, and 160mg/m3 Statistical methods used: Body weight, food consumption, hematology and clinical chemistry parameters, organ weights, and organ/body weight ratios were analyzed. Mean values of all dose groups were compared to control at each time interval . Tests included parametric ANOVAwith a Dunnett's post-hoc test, non-parametric Kruskal-Wallis and Dunn's rank sum test, Bartlett's test for equal variances, and Student's t-test . Test method/conditions remarks: The rats were exposed for 6 hours/day, 5 days/week for 4 weeks to the test material (target concentrations = 15, 50, and 150 mg/m) as a liquid droplet aerosol generated by an air atomizing nozzle apparatus delivered into a plexi-glass chamber. Control rats were exposed in the same manner as the test-material-exposed group except that mineral oil only was administered at a level of 150 mg/m3 . The details of the whole-body

42Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

exposure are consistent with those described in OECD TG 412 . The actual exposure concentrations, as measured by gravimetric analysis, were 15, 50, and 160 mg/m3 . Particle size analyses were performed once/week fromthe test material chamber using a cascade impactor. Animal observations for toxicological signs and mortality were recorded twice daily, once in the morning and once in the afternoon. Individual weights were recorded twice pre-test and then weekly during the exposure and recovery periods, and at termination. At the conclusion of the observation period, the surviving animals were euthanized with carbon dioxide. Animals were fasted prior to sacrifice. Five rats/sex were subjected to post-exposure blood analysis (routine hematology and clinical chemistry parameters) on test day 1 for thecontrol and high-dose groups, at termination on 5 rats/sex for all dose groups, and on 5 rats/sex fromthe control and high-dose group after threeweeks ofrecovery. Complete gross post mortemexaminations were performed on all animals (nasal passages, trachea, external surface, all orifices, the cranial cavity, the brain and spinal cord, and allviscera) . Nine major organs were weighed to obtain organ/body weight calculations, 42 individual organs and/or tissues were preserved, and 10 major organs and/or tissues were examined for histopathology. Endpoint value(s) : NOAEL (male) = 160mg/m3 based on no relevant effects at the highest dose tested; LOAEL (female) = 160 mg/m3 based on increased hemoglobin levels; NOAEL (females) = 50 mg/m3 Results remarks: The mass median aerodynamic diameter for the studies ranged from1 .9 to 2.6 microns with a geometric standard deviation ranging from1 .8 to 2.2. These data indicated that the aerosol was of a respirable size in the rat, with at least 96% ofthe particles 10 microns or less in diameter . Mortality: One high-dose female had convulsive behavior following the third dayof exposure, andwas found dead the next morning. The cause ofdeath was unclear. There were no other unscheduled deaths in the study. Physical observations: The animals were unremarkable during the exposure period. Weekly detailed observations included an increased incidence ofnasal discharge or dried red material on the facial area amongthe high-dose animals. However, these findings were not temporally consistent nor were they apparent in the lowest two doses of test material . No significant respiratory sounds were noted. Body weights: Although there were no significant differences seen between control and treated groups, there was a trend toward lower body weight gains during the exposure period ofthe study at all dose levels in the males and with the two highest dose levels in the females. During the three-week recovery period, the high-dose animals did not regain the difference in body weight compared to the controls. Hematology: The only significant difference fromcontrol values was increased hemoglobin concentration in the high-dose females sacrificed after 4weeks of exposure.Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

Clinical chemistry: There were several statistically significant differences fromthe control values at both the post-exposure and post-recovery time intervals. However, these differences did not correlate with dose, with sex, with potential target organs, or with sacrifice interval . Terminal organ and body weights: Following 4weeks of exposure to test material, increases in kidney weights were seen in the males at all three dose levels, and were statistically significant in the higher two levels. This effect was considered to be related to renal effects seen microscopically in males (see below) . This difference in weight abated following 3 weeks of recovery. Following 4weeks of exposure, statistically significant increases were seen in highdose liver weights and liver/body ratio in both sexes. These differences abated following 3 weeks of recovery. Spleen and adrenal weights increased compared to controls in the high-dose groups ofboth sexes. Post-recovery increases in testes, heart, lung, and spleen weights were recorded. These effects were not accompanied by pathologic microscopic findings, and therefore, the biological significance was considered equivocal . Afew visible gross changes, such as discolored lungs, were noticed in the sacrificed animals. Microscopically, treatmentrelated effects were seen in the kidneys in the males in a dose-related profile. Findings included globular casts at the corticomedularyjunction, the cortex and medulla, as well as hyaline droplets in the proximal convoluted tubule cells. These responses were seen in males in all treatment groups following 4weeks of exposure, and in the high-dose group after 3 weeks of recovery. The nephropathy in males is consistent with the alpha 2p-globulin-mediated mechanism that is male rat-specific and is not considered by EPAto be predictive of comparable toxicity in humans. All other microscopic tissue alterations observed in other organs were considered incidental findings. CONCLUSIONS The LOAEL for females is 160mg/m3 based on increased hemoglobin levels in females. The NOAEL is 50 mg/m3 in females. The NOAEL for males is 160 mg/m3 based on no human health-related effects at the highest dose tested. STUDY RELIABILITY [1] Reliable without restrictions. REFERENCES Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of theAmerican Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 5 : Repeated-dose dermal toxicity study of CASRN68515-88-8 in rats Title: Not available This study is : ®

Adequate F Not Adequate TEST SUBSTANCE Identity : Pentene, 2,4,4-trimethyl-, sulfurized, CASRN 68515-88-8 Remarks: Purity not provided METHOD Method/guideline followed: Consistent with OECDTG 410and OPPTS 870 .3200 Test type: 28-Day Dermal Toxicity Study in Rats GLPcompliant?: Yes Year study performed: 1988 Test species: Sprague-Dawley CDrat (21 days old at initiation of treatment) Number of animals/sex/dose: 5 males/dose Route of administration: Dermal Vehicle : None Exposure duration: 5 days/week, 4weeks Doses/concentrations tested: 0 or 1000 mg/kg/day Statistical methods used: Continuous data including body weight, body weight gain, and food consumption were analyzed by analysis ofvariance. Test method/conditions remarks: Rats were administered the test substance at 0 or 1000 mg/kg/day topically on shaved, unabraded skin under semi-occluded conditions for 6hours/day, 5 days/weekfor 4weeks (20applications) . Controls rats received topical application of mineral oil . Following the exposure period, the test material was removed with mineral oil . Endpoint value(s) : NOAEL = 1000 mg/kg/day based on no systemic toxicity at the highest dose testedFinal HPV3 Test Plan and Data Adequacy Review -_ CASRN 72162-15-3

Results remarks : All animals survived throughout the study and physical examinations were generally unremarkable. No difference between the test material-treated animals and control animals was noted for the parameters ofbody weight, body weight gain, or food consumption. Detail gross pathological examination of external and internal features ofthe animals revealed no remarkable findings with the exception of weak-moderate irritation responses at the site oftest material application. These responses were characterized by erythema, eschar, and flaking of the skin that persisted over the majority of the 28-day treatment period. CONCLUSIONS All animals survived throughout the study and physical examinations were generally unremarkable. The NOAEL is 1000 mg/kg/day (only dose tested) . STUDY RELIABILITY [2] Reliable with restrictions. Data for clinical chemistry, hematology, and histopathology are not presented. REFERENCES Reference information is not available (the underlying study contains confidential business information) . This studywas obtained fromthe Robust Summaries for theAlkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

C-7. Genetic Toxicity - Bacterial Reverse Mutation Summary of endpoint : Adequate Study 1 : Mutagenicity study ofCASRN685I1-50-2 in Salmonella typhimurium Title : Not available This study is: ®

Adequate 0Not Adequate TEST SUBSTANCE Identity : 1-Propene, 2-methyl- sulfurized, CASRN68511-50-2 Remarks: Purity not provided 

### METHOD Method/guideline followed: Consistent with guidelines outlined in OECDTG 471 Test type: Bacterial Reverse Mutation Assay GLPcompliant? : No Year study performed: 1978 Test species: Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, and TA98 Metabolicactivation : With and without Concentrations tested: 0, 0 .01, 0 .05, 0 .1, 0.5, 1 .0 gL/plate Statistical methods used: Determination ofmean t S.D . ofreplicate plate counts Test method/conditions remarks: The vehicle was DMSO . All stock and working solutions were stored at 4°Cin glass screw-capped bottles. All sterility controls were negative for bacterial growth. Vehicle was tested as negative control . Activation consisted of adult male Sprague-Dawley rat liver S-9fraction, inducedwith Aroclor 1254 (100 gl/plate) . RESULTS Genotoxicity result : Not mutagenic Cytotoxic concentration (if applicable) : Not specified Results remarks : For all strains and dose levels with and without metabolic activation, the criteria for a positive mutagen (at least 3 times the number of colonies as the controls for 47Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

spontaneous reversion) were not met . Positive controls (9-aminoacridine and 2-nitrofluorene without activation and 9-aminoacridine, 2-nitrofluorene, aflatoxin, and 6-aminochrysene with activation) had at least 3 times the number of colonies as the control . CONCLUSIONS Thetest chemical does not induce gene mutations in bacteria either with or without metabolic activation. STUDY RELIABILITY [2] Reliable with restrictions. Cytotoxicity was not noted and the assaywas conducted using very low concentrations. REFERENCE Reference information is not available. This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Programby the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum

Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 2: Mutagenicity study ofCASRN685I5-88-8 in Salmonella typhimurium Title: Not available This study is: ®

Adequate F Not Adequate TEST SUBSTANCE Identity: Pentene, 2,4,4-trimethyl-, sulfurized CASRN 68515-88-8 Remarks: 97%

purity METHOD Method/guideline followed: Consistent with guidelines outlined in OECDTG 471 Test type: Bacterial Reverse Mutation Test GLPcompliant? : Yes Year study performed: 1982 Test species: Salmonella typhimuriumstrains TA98, TA100, TA1535, TA1537, and TA1538 Metabolic activation: With and without Concentrations tested: 0, 0 .01, 0 .03, 0 .1, 0 .3, and 1 1tL/plate Statistical methods used: The mean number ofhis' revertants/plate for three replicate assay plates was calculated for each concentration and strain. Test method/conditions remarks: No significant deviations fromguideline protocols. The vehicle was DMSO

. RESULTS Genotoxicity result : Negative with and without metabolic activation. Cytotoxic concentration (if applicable) : Not observed Results remarks : The test material was tested without metabolic activation at 1, 0 .3, 0 .1, 0 .03, and 0.01 1xL of test material/plate and found to be non-mutagenic to the bacterial strains tested. The number of revertant colonies as a result of treatment with the test material did not differ significantly fromthe number produced by the DMSOvehicle control . The test material was not toxic to any strain at any concentration. Thepositive controls, sodium azide, 2-nitrofluorene, and 9-aminoacridine at concentrations ranging from2.5-100 pg/plate produced more than a 10- fold greater incidence of his+ revertants/plate with the bacterial strains used in this study. The 49Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

test material was also tested in the presence of an S9 microsomal fraction from Aroclor 1254- treated rat livers. The concentrations tested (1, 0 .3, 0 .1, 0 .03, and 0

.01 1xL of test material/plate) in theactivated systemdid not induce significant detectable mutagenic events with the bacterial strains used. The positive metabolic activated control, 2-anthramine (2.5 ltg/plate) produced positive mutagenic responses in the bacterial strains used in this study. CONCLUSIONS Thetest material did not produce significant mutation in any ofthe Salmonella strains in the quantitative mutagenesis assay, either in the presence or absence of metabolic activation. Thus, under the conditions of the assay employed, the test material was determined to be nonmutagenic in the Salmonella/microsome mutagenesis assay . STUDY RELIABILITY [2] Reliable with restrictions. Cytotoxicity was not observed and the assay was conducted using very low

concentrations . REFERENCE Reference information is not available (the underlying study contains confidential business information) . This studywas obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Programby the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Study 3: Mutagenicity study ofCASRIV 67762-55-4in Salmonella typhimurium Title: Not available This study is: ®

Adequate F Not Adequate TEST SUBSTANCE Identity: Alkenes, C15-18 alpha-, sulfurized, CASRN 67762-55-4 Remarks: Purity not provided METHOD Method/guideline followed: Designed to be in compliance with microbial mutagenicity testing as set forth by OECD 1981, EPA 1982, FDA 1993 Test type: Bacterial Reverse Mutation Test GLP compliant?: Yes (except analyses were not performed to verify the homogeneity, stability or accuracy ofthe test/control article preparation) Year study performed: 1996 Test species : Salmonella typhimuriumstrains TA1535, TA1537, TA98, TA100, and TA102 ; Escherichia coli strain WP2uvrA Metabolic activation: With andwithout Concentrations tested: Pre-screen, duplicate cultures: 50.0, 167, 500, 1670, and 5000 pg/plate, plus control ; Triplicate cultures: 50.0, 167, 500, 1670, 5000, and 10,000 wg/plate Statistical methods used: Statistical analyses are performed using the program developed by Snee and Irr (1981), with significance established at the 95% confidence limit. Test method/conditions remarks : Test articlewas first evaluated in a pre-screen using both liquid preincubation and plate incorporation treatment conditions. Duplicate cultures of strains TA1537, TAI00, and WP2uvrA were treatedwith article at doses of 50 .0, 167, 500, 1670, and 5000 Wg/plate, as well as the solvent control, in the absence of S9 . The test article was found to be incompletely soluble (droplets were observed) at all doses. The article was next evaluated using both treatment conditions. Based upon the results of the pre-screen, the article was evaluated in triplicate cultures in strains TAI 535, TA1537, TA98, TA100, TAI02, and WP2 uvrA in the presence and absence of S9 at doses of 50.0, 167, 500, 1670, 5000, and 10,000 pg/plate . Six doses of the article were evaluated in the event of unacceptable toxicity and/or insolubility at the highest dose levels evaluated in the mutation assay. The S9 mixture included 6%(v/v) Aroclor 1254-induced male Sprague-Dawley rat liver homogenate with the appropriate

51Final HPV3 Test Plan and Data Adequacv Review CASRN 72162-15-3

buffer and cofactors. The test article was again found to be incompletely soluble at all doses, under both treatment conditions. RESULTS Genotoxicity result: Negative with and without metabolic activation Cytotoxic concentration (if applicable) : Not specified Results remarks: In the pre-screen, results indicated that the article was not toxic. In the following study, normal growth was observed in all tester strains at all doses evaluated with and without S9. Revertant frequencies for all doses of article in all tester strains, with and without S9 under both treatment conditions approximated or were less than those observed in the concurrent negative control cultures. All positive and negative controls were within acceptable ranges. CONCLUSIONS The results were negative in this study, using liquid pre-incubation and plate incorporation treatments. STUDY RELIABILITY [2] Reliable with restrictions. No analyses were supplied to verify the homogeneity, stability, or accuracy of test article preparation. The test article was found to be incompletely soluble (droplets were observed) at all doses, under both treatment conditions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This studywas obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data A dequacy Review CASRN72162-15-3 C-8 . Genetic Toxicity - Chromosomal Aberrations Summary of endpoint : Adequate Study 1 : Micronucleus assay ofCASRN68511-50-2 in mice Title: Not available This study is: ®

Adequate 0 Not Adequate TESTSUBSTANCE Identity : 1-Propene, 2-methyl, sulfurized-, CASRN68511-50-2 Remarks: Purity not provided METHOD Method/guideline followed: OECD TG 474 Test type: Mammalian Erythrocyte Micronucleus Test (in vivo) GLPcompliant?: Yes Year study performed: 1989 Test species: B6C3F1 mice Number of animals/sex/dose: 15/sex/dose (5/sex/dose/sample interval) Route of administration : Intraperitoneal Vehicle: Hydroxypropyl methylcellulose (Methocel K4M Premium

- Dow Chemical) Exposure duration : Single dose Doses tested : 3500 mg/kg bw Statistical methods used : Normal test for equality ofproportion (one-tailed) . Because of multiplicity of comparisons, a Dunnett adjustment was made. Test method/conditions remarks : Young male and female mice were treated with a single intraperitoneal injection of3.5 g/kg test material, 50 mg/kg cyclophosphamide in saline, or methylcellulose vehicle alone. Dose had been determined in a preliminary toxicity test to identified MTDfor this study . Animals were sacrificed and femurs removed at 24, 48, or 72 hours post-dosing (5M, 5F per interval) for test material and negative control, andat 24 hours

53Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

post-dosing only for cyclophosphamide. Bone marrowsmears were prepared and immature red blood cells (polychromatic erythrocytes, PCEs) and mature red blood cells (normochromatic erythrocytes, NCEs) were evaluated for toxicity andthe presence of micronuclei . Slides were stainedwith acridine orange and scored under a fluorescence microscope. Slides from all dose groups were sorted by a computerized randomnumber systemand the cytogeneticist was unaware of what dose group any individual slide was from . The ratio of PCE or NCE per the first 1000 erythrocytes counted was calculated to determine cytotoxicity, if any. RESULTS Genotoxicity result : Negative Cytotoxic dose: Not observed Results remarks : In the preliminary toxicity test (2M, 2F/group) all mice died at 5.0 g/kg and all survived at 3.5 g/kg with no cytotoxicity in bone marrowcells 24 hours after injection. Data fromthe full study demonstrate that the frequency of mironucleated PCEs in femoral bone marrowfor males and females treated with the test material was not significantly elevated (p < 0 .05) when compared to negative controls for groups sampled at 24, 48, or 72 hours postinjection. Results fromboth sexes combined demonstrate the same results. Cyclophosphamide, the positive control material, did induce statistically significant increases in micronucleated PCEs in all animals demonstrating that avalid studywas performed. CONCLUSIONS Methyl propene derivative administered via i .p. injection at 3.5 g/kg body weight did not induce the formation of micronuclei in PCEs in male or female mice at anytime interval and is not considered clastogenic in this test system . STUDY RELIABILITY [I] Reliable without restrictions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's PetroleumAdditives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adecauacv Review CASRN72162-15-3

Study 2 : Micronucleus assay of CASRN68511-50-2 in rats Title: Not available This study is: ®

Adequate F~ Not Adequate TEST SUBSTANCE Identity: 1-Propene, 2-methyl-, sulfurized, CASRN 68511-50-2 Remarks : Purity not provided METHOD Method/guideline followed: Modified OPPTS 870 .5395 Test type: Mammalian Erythrocyte Micronucleus Test (in vivo) GLPcompliant? : Yes Year study performed: 1987 Test species: Sprague-Dawley (Tac :N[SD]fBR) rats Number of animals/sex/dose: 5/sex/dose Route of administration: Dermal Vehicle: Mineral oil (solvent refined naphthenic base stock) ; density 0 .88 g/L Exposure duration: 5 days/week for 13 weeks Doses tested : 500or 2000mg/kg/day undiluted test material ; 500mg/kg/daydiluted 50% w/v in mineral oil base stock Statistical methods used: SAS ANOVAand ANOVA F test ; Tukey's Studentized Range test and Scheffe's test ; SAS General Linear Model, a studentized linear regression analysis to determine dose responsiveness. Statistical analyses compared test values to negative control data; a significant increase in micronuclei is an indicator of clastogenic activity by the test material Test method/conditions remarks : Age at initiation: 7 weeks old following 2weeks acclimation .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

Methyl propene derivative was applied to the clipped backs of groups of 20 Sprague-Dawley rats (1 OM, l OF) 5 days per week for 13 weeks at dose levels of0, 500, or 2000 mg/kg/day undiluted or 500mg/kg diluted (50% w/v) in mineral oil base stock. Rats were fitted with Elizabethan collars to minimize ingestion of test material, which was left uncovered on the skin. At termination ofthe 13 week subchronic study, approximately 24 hours after the final dermal administration, bone marrow was harvested from

femurs of the first 5 rats/sex/group necropsied. Three bone marrowslides were prepared for each animal . Slides were stained with acridine orange and scored under afluorescence microscope. All slides were randomized by a computergenerated randomnumbers table so that the cytogeneticist was unaware of what dose group any individual slide was from . Immature red blood cells (polychromatic erythrocytes, PCE) and mature red blood cells (normochromatic erythrocytes, NCE) were evaluated for toxicity and the presence of micronuclei . The ratio of PCE to NCE per the first 1000 erythrocytes counted was calculated to determine cytotoxicity if any . At least 1000 PCE and 1000 NCE were scored for the presence of micronuclei . RESULTS Genotoxicity result : Negative Cytotoxic dose: No data Results remarks : Methyl propene derivative undiluted (500 or 2000 mg/kg/day) and methyl propene derivative (500 mg/kg/day) 50% w/v in mineral oil base stock were not cytototoxic to red blood cell formation. Thesetest materials did not induce any statistically significant increase in theformation of micronucleated PCEs or NCEs in bone marrow red blood cells ofmale or female rats exposed dermally for 13 weeks. CONCLUSIONS Methyl propene derivative does not cause chromosome damage in rats following regular and prolonged dermal exposure in this test system . NOEL= 2000 mg/kg/day STUDY RELIABILITY [2] Reliable with restrictions. Study does not include a positive control . Samples were collected only once approximately 24 hours after the final dose. Information on test material composition, purity, and stability are not part of this report, but the reader is referred to the adjunct 13-week subchronic report . REFERENCE Reference information is not available (the underlying study contains confidential business information) . This studywas obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) . 56Final HPV3 Test Plan and Data Adequacv Review CASRN 72162-15-3

Study 3 : Micronucleus assay of CASRN68515-88-8 in mice Title: Not available This study is: ®

Adequate F-1 Not Adequate TEST SUBSTANCE Identity: Pentene, 2,4,4-trimethyl-, sulfurized, CASRN 68515-88-8 Remarks: 97%

purity METHOD Method/guideline followed: OECDTG 474 Test type: Mammalian Erythrocyte Micronucleus Test (in vivo) GLP compliant? : Yes Year study performed: 1988 Test species: B6C3F1 mice Number of animals/sex/dose : 5/sex/dose Route of administration: Oral gavage Vehicle: Not indicated Exposure duration: Single dose Doses tested: 5000 mg/kg Statistical methods used: Group mean body weights, total polychromatic erythrocytes (PCEs), normochromatic erythrocytes (NMEs), PCEs with micronuclei, and NMEs with micronuclei were compared. For each animal, a minimum of 1000 PCEs were counted for the presence of micronucleated PCEs. The frequency of micronucleated cells per animals was expressed as the number ofmicronucleated PCEs per 1000 PCEs counted. The ration of PCEs/NMEs was also recorded. The data were analyzed for statistical significance on a binomial distribution, at a level of significance of0

.05, and using the table of Kastenbaum and Bowman (Mutation Res. 9 :527- 549,1970) . Test method/conditions remarks: Number of animals per dose: 5/sex/group. 57Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

Control groups and treatment: 5/sex negative control (mineral oil) ; 5/sex positive control (cyclophosphamide, 50 mg/kg intraperitoneal injection) . Mice were approximately 12 weeks old and 17-31 grams at study initiation. Animals were observed daily and body weights were recorded after 18, 24, and 48 hours. Test material and negative control groups were sacrificed after 18, 24, and 48 hours, whereas the positive control group was terminated after 24 hours. RESULTS Genotoxicity result : Negative Cytotoxic dose: No data Results remarks: The frequency of PCEs with micronuclei ranged from1 .0 to 5.9/1000 PCEs in negative control mice with groups means of2.6, 3.0, and 2.4 PCEs for the three time points. These averages and group meanswere within the expected range based on published data on the performing laboratory historical controls. In contrast, male animals dosed with cyclophosphamide had 9 .0 to 24 .0 micronucleated PCEs/1000 PCEs, with a mean of 14.5 for the group. The average frequencies of micronucleated PCEs obtained frommale animals receiving the test material after the three time periods were 5.1, 3.0, and 5.7/1000 PCEs. These group means were not significantly higher than the negative control values. The mean PCE/NME ratios in negative male group for the three time periods were 0 .60, 0 .60, and 0

.69, respectively. Thetest material was not cytotoxic since the PCE/NME ratio at the three time points was 0 .60, 0 .59, and 0

.66 . The mean frequency of micronucleated PCEs/1000 PCEs for female mice was 1 .9, 2.1, and 2.9, respectively. The average micronucleated PCEs value for the cyclophosphamide treated females was 20 .5. Female mice treated with the test material were foundto have mean micronucleated PCEs values of 1 .1, 2.0, and 1 .2 at the three time points, respectively. Acomparison ofthe PCE/NME ratio between the negative control and test material treated female mice did not vary significantly. CONCLUSIONS The subject material was tested for its genotoxicity using mouse in vivo micronucleus screening assay in bone marrow

. There was no significant increase in micronucleated PCEs in animals exposed to the test substance. Thus, the test material was negative in this assay . STUDY RELIABILITY [1] Reliable without restrictions. REFERENCE Reference information is not available (the underlying study contains confidential business information) . This study was obtained fromthe Robust Summaries for the Alkyl Sulfides Category (submitted to EPAunder the HPVChallenge Program by the Health, Environmental, 58Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

and Regulatory Task Group of the American Chemistry Council's Petroleum Additives Panel on May 19, 2005) .Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

C-9 . Reproductive Toxicity Summary of endpoint : Not adequate No studies were available for this endpoint .Final HPV3 Test Plan and Data Adequacy Review CASRN72162-15-3

C-10 . Developmental Toxicity Summary of endpoint : Not adequate No studieswere available for this endpoint .Final HPV3 Test Plan and Data Adequacy Review CASRN 72162-15-3

D . DATA ADEQUACY SUMMARY TABLE Endpoint Study Information Available Acceptable Data Gap Acute Toxici ty to Fish Yes Yes No Acute Toxici ty to Dap

hnia Yes Yes No Chronic Toxici ty to Dap hnia No Yes Toxicity to Plants (Algae) Acute Toxicity Yes Yes Yes

Yes No

No Rep eated-Dose Toxicity Yes Yes No Gene Mutation Yes Yes No Chromosomal Aberrations Yes Yes No I Rep

roductive Toxicit y No Yes Developmental Toxicity No - YesANGELAJACKSON 8007536537 DOVER CHEMICAL 3676 DAVIS ROAD NW DOVER OH 44622 SHIP TSCASECTION4l2021564-8930 TO : DOCUMENT CONTROL OFFICE 1200 PENNSYLVAINAAVENUE NW ENVIRONMENTAL PROTECTIONAGENCY LTR 7 OF 1 

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