### Testing of Certain High Production Volume Chemicals; Thir d Group of Chemicals Final Rule

RESPONSE TO PUBLIC COMMENTS

Prepared by the Chemical Information and Testing Branch, Office of Pollution Prevention and Toxics, Office of Chemical Safety and Pollution Prevention, U.S. Environmental Protection Agency September 12 , 20112 T 

### able of Contents Unit Title Page A . Introduction 4 

### B. General Comments and t he Use of Existing Information 5 C. Comments on Proposed Test Methods 5 1. Acute toxicity test method 5 2. Aquatic toxicity test method 7 3. Genetic toxicity test method 9 4. Reproductive and developmental toxicity screening 9 D. Animal We lfare Considerations 11 E. Chemic al Specific Comments 12 1. Benzene, 1,2 - dimethyl - 3 - nitro - (CASRN 83 - 41 - 0) 12 2. 3 - Pentanone ( CASRN 96 - 22 - 0) 14 3. Benzene, 1 - chloro - 4 - (trifluorom ethyl) - (CASRN 98 - 56 - 6) 15 4. Benzenesulfonic acid, 3 - nitro, sodium s alt (1:1) (CASRN 127 - 68 - 4) 17 5. 1 - Tetracosanol (CASRN 506 - 51 - 4); 1 - Hexac osanol (CASRN 506 - 52 - 5) 19 6 . 2 - Propenoic acid, 2 - carboxyethyl ester (B - CEA) (CASRN 24615 - 84 - 7 25 7 . Benzenesulfonic acid, dimethyl - (CASRN 25321 - 41 - 9) 26 8. Methanesulfonamide, N - [2 - [(4 - amino - 3 - methylphenyl)ethylamino]ethyl] - , sulfate (2 :3) (CASRN 25646 - 71 - 3) 31 9 . 1 - Propanesulfonic acid, 2 - hydroxy - 3 - (2 - propen - 1 - yloxy) - , sodium salt (1:1) (CASRN 52556 - 42 - 0) . 343 1 0 . Coal/Coal Tar Chemical Sub stances: 35 Solvent naphtha (coal) (CASRN 65996 - 79 - 4); Tar oils, coal (CASRN 65996 - 82 - 9); Tar, coal, high - temperature (CASRN 65996 - 89 - 6); Distillates (coal tar) (CASRN 65996 - 92 - 1); Pitch, coal tar - petroleum (CASRN 68187 - 57 - 5); Extract residues (coal), tar oil alk., naphthalene distn. resi dues (CASRN 73665 - 18 - 6) 1 1 . 1,4 - Benzenedicarboxylic acid, 1,4 - dimethyl ester, manuf. of, by - products from (CASRN 68988 - 22 - 7) 48 1 2 . 1 - Decene, sulfur ized (CASRN 72162 - 15 - 3) 5 3 Bibliography 5 64 A. Introduction This document contains a summary of the comments received by EPA in response to the TSCA section 4 proposed test rule entitled “Testin g of Certain High Production Volume Chemicals, Third Group of Chemicals” (75 FR 8575, February 25, 2010) along with EPA’s responses to those comments. The proposed rule and comments are available in EPA’s public docket and e - docket for the final test rule under Docket No . EPA

– HQ

– OPPT

– 200 9

– 0 112 . EPA received comments from industry, including:

**•** American Chemistry Council (ACC) **•** The American Coke and Coal Chemicals Institute (ACCCI) **•** BASF **•** Chlorinated Paraffi ns Industry Association (CPIA) **•** Eastman Chemical Company (Eastman) **•** Greenwich Chemical Consulting, Inc. (GCC) **•** International Imaging Industry Association (I3A) **•** INVISTA **•** IsleChem, LLC (IsleChem) **•** Rhodia, Inc. (Rhodia) **•** Sasol North America ( SASOL ) **•** GE Water and Process Technologies (GE) **•** Special Materials Company (SMC) Greenwich Chemical Consulting, Inc. (GCC) submitted comments on behalf of Brenntag North America, Inc.; Kowa American Corporation; Miami Chemical, Inc.; and Univer USA, Inc. (“The Companies”). EPA also received comments from a number of Animal W elfare Organizations (AWOs) . AWOs that submitted comments under the letterhead of People for the Ethical T reatment of Animals (PETA) included PETA, the Physicians Committee for Responsible Medicine (PCRM), the Alternatives Research Development Foundation, and the American Anti -5 Vivisection Society . EPA also received a comment from a private citizen concerned with companies using animals to test their commercial products and advocated other (non - animal) tests for this purpose. B. General Comments and the Use of Existing Information Comment . AWOs commented that, in cases where chemical properties and environmental fate data are lacking, no toxicity tests should be conducted until after such basic testing is completed, as certain chemical/fate properties may p reclude the need to conduct toxicity tests. Response . While it is true that physical chemical property and fate data can be useful in determining how specific toxicity tests are performed, few chemical and fate properties negate the requirement for testing altogether . Those physical properties that are most related to bioavailab i lity, the water solubility and octanol - water partition coefficient, are readily estimated using chemistry modeling computer programs, should these data be necessary for experimental design. With the exception of intrinsic chemical rea ctivity, knowledge of a chemical’s environmental fate is generally not necessary prior to assessing its toxicity . It is agreed that highly reactive chemicals that undergo instantaneous destruction upon exposure to the environment should not be tested for t oxicity . However, reactivity of this nature is generally well known to those who manufacture, transport, use or otherwise handle the chemical in question . In addition, the reactivity of the parent does not necessarily preclude testing of the degradation pr oducts . Therefore, unless there is a question regarding the reactivity of the chemical, there is no reason to withhold toxicity testing until all of the chemical/fate property testing has been conducted. C . Comments on Proposed Test Methods 1. Acute toxi city test method. Comment . AWOS commented that the acute toxicity testing requirement may not be necessary. According to AWOs, evidence indicates that more than 85% of industrial chemicals are non -6 toxic (Kinser - Ovaskainen et al., 2008) and efforts to vali date the in vitro 3T3 NRU cytotox assay ( ICCVAM , 2006) also indicate that this assay correctly discriminates non - toxic chemicals from toxic ones . AWOs suggest that EPA wait for the final results of this validation before requiring in vivo (acute mammalian) toxicity testing, which is specified in the test rule. AWOs stated that a coalition of European pharmaceutical companies determined that regulatory decisions were almost never predicated on the results of acute toxicity tests, which prom pted the International Council on Harmonization and its regional pharmaceutical regulators to propose removing the requirement for an acute oral toxicity testing from the testing guidelines (Robinson et al., 2008; European Medicines Agency, 2008) and obtai n necessary information on acute toxicity from available repeated dose studies . According to AWOs, the European Center for the Validation of Alternative Methods (ECVAM) used the approach of the International Council on Harmonization and correctly classifie d a majority of chemicals (Kinsner - Ovaskainen et al., 2008) . Additionally, AWOs state that ECVAM is currently conducting a definitive validation of this hypothesis with thousands of chemicals . AWOs also suggested that an emerging method to estimate toxicit y for exposures via the “inhalation route” (Mackay et al., 2008; Veith et al., 2008) should also be pursued . A similar comment was made by AWOs in response to the TSCA section 4 proposed test rule entitled “Testing of Certain High Production Volume Chemica ls, Second Group of Chemicals” (73 FR 43314, July 24, 2008). Response . While EPA generally agrees that, for a variety of reasons, test methods which minimize the use of live animals are desirable, a new in vitro method, such as one that would estimate toxicity for exposures via the “inhalation route,” must be validated before in vivo testing can be partially or completely eliminated . In the case of the 3T3 NRU cytotoxicity assay, EPA agrees with the conclusion of a 2006 ICCVAM Test Method Evaluation Report (ICCVAM, 2006); namely, that data from the 3T3 NRU cytotoxicity assay may be used in a weight - of - evidence approach, where appropriate, for determining starting doses for in vivo acute oral systemic toxicity stud ies (EPA 2008b). EPA acknowledges that this use of this approach may result in the reduction of animals used in acute oral toxicity testing . As discussed below, EPA also agrees with ICCVAM’s conclusion that these in vitro test methods are not7 sufficiently validated to replace animals for regulatory labeling and hazard classification purposes (EPA 2008b). Although EPA agrees that the 3T3 NRU assay can be used in a weight - of - evidence approach, it should be acknowledged that the conclusions of the ICCVAM re port, as well as an OECD guidance document on the use of the 3T3 NRU cytotox assay were based on validation studies conducted on only 72 chemicals . EPA agrees that an evaluation of thousands of chemicals using the 3T3 NRU cytotoxicity assay would be more d esirable in determining the true validity of the assay. However, t he results of the validation by ECVAM using thousands of chemicals are not available for EPA to evaluate as of this writing. It should also be noted that the 3T3 NRU cytotox assay is for the purpose of estimating starting doses -- not as a complete replacement of in vivo testing -- for acute oral systemic toxicity tests. Although the 3T3 cytotox assay is not currently accepted as a substitute for in vivo testing, EPA allows the use of in vitro testing and similar approaches, such as Structure - Activity Relationship ( SAR ) models, to estimate the likelihood of toxicity of chemicals and thereby reduce the number of animals necessary to determine toxicity. 2. Aquatic toxicity test method . C omment . AWOs stated that in vivo aquatic toxicity testing is unnecessary considering the availability of suitable in silico and in vitro test methods such as ECOSAR and the DarT test. Response . At the current time, the DarT test is not considered to be a dequately validated to use as a replacement for acute fish toxicity testing (see response to next comment below) . ECOSAR modeling, while useful in estimating toxicity, is not considered to be an acceptable method for the definitive determination of toxicit y of chemicals for regulatory purposes . Comment . AWOs commented that the DarT test, using fertilized zebrafish eggs as a surrogate for living fish, has been nominated for development into an OECD test guideline and should be used as a replacement for (ac ute) fish toxicity testing . If fish acute toxicity testing is still required, a threshold (step - down) test (e.g., Jerama et al., 2005) should be used, with an8 additional refinement also possible to terminate testing after 24 hours of exposure in some cases (Hutchinson et al., 2003) . AWOs stated that the Fish Threshold Approach (FTA) is a modification of the step - down approach and that an OECD Guidance Document describing this approach is in the process of being finalized (OECD, 2010). Response . EPA notes that the DarT test has received attention as a potential in vitro replacement for the current acute fish toxicity testing (e.g., Brannen et al., 2009). However, this test method is still being refined and undergoing validation. U ntil the DarT test has been adequately validated, the Agency will continue to recommend approved guidelines, rather than methods that are still under development, which may in fact not be adequately predictive. The acute fish toxicity test conducted in vivo will continue to be requi red. Acute aquatic toxicity testing determines a 96 - hour LC 50 ; i.e., the concentration at which 50% mortality is observed after 96 hours of exposure to a chemical. The Hutchinson et al. (2003) method appears to suggest that testing be terminated at 24 hours of exposure when lethality and/or serious morbidity are observed in two out of five fish (as described by AWOs) . The authors state that this method could be used to interpolate an LC 50 value . While this may be useful for estimation purposes, t his met hod of extrapolation is not acceptable to EPA for determining an LC 50 . Comment . AWOs suggested that toxicity testing should be conducted first on aquatic plants and invertebrates, as toxicity to these organisms (fish food supply) might obviate the need for fish toxicity testing. Response . EPA disagrees. Aquatic plants and invertebrates are included with fish in the evaluation of aquatic toxicity in order to determine the toxicity of a chemical to three different taxonomic levels of the aquatic ecosystem . In addition, it is possible that a chemical substance may be much more toxic to fish than to aquatic plants or invertebrates . Therefore, it is essential that all three tests be performed to adequately assess the risk to aquatic organisms.9 3. Genetic toxicity test method . Comment . AWOs suggested that the genetic toxicity sections of the proposed rule should be amended to mandate (rather than encourage) the use of in vitro genetic toxicity testing; with the requirement that any company proposing to us e an in vivo method first provide a compelling justification for an exemption from the in vitro test methodology, with opportunity for public comment . AWOs stated that in response to comments on the first proposed test rule, EPA anticipated that test spons ors would perform less expensive in vitro tests unless there were sound scientific reasons not to do so . However, AWOs indicated that retrospective analysis of all test plans submitted to the HPV Challenge Program indicated that only 21% of in vivo chromosomal aberration tests originally proposed were subsequently replaced by in vitro test s in revised test plans. Response . In the proposed rule, and in its comments to test sponsors, EPA has made it clear that in vitro testing is preferred for collec ting screening data for genetic toxicity endpoints . In the HPV Challenge Program and in the proposed rule, EPA has asked sponsors who submit test plans for in vivo genetic toxicity testing to include justification for the in vivo method instead of the in v itro method . EPA does not mandate that genetic toxicity testing be conducted in vitro in the HPV program because, as stated in the proposed rule, chemical properties may preclude use of the in vitro method . Also, extant in vivo genetic toxicity data for an HPV chemical or a supporting HPV chemical compound may be adequate to satisfy the genetic toxicity endpoints so that further testing is not required . In cases where an HPV chemical requires genetic mutation or chromosomal aberrations testing, EPA permits the use of in vivo testing, but only after those providing the testing present a strong justification for in vivo testing in lieu of in vitro testing . 4. Reproductive and developmental toxicity screening. Commen t . AWOs commented that EPA should amend the proposed rule to require companies to use the combined repeated - dose toxicity study with the reproduction/developmental toxicity screen ing test unless, before testing, a compelling justification for an exemption fr om the10 combined protocol is provided, with opportunity for public comment available . AWOs stated that in response to comments on the first proposed test rule, EPA anticipated that test sponsors would perform less expensive combined tests unless there were sound scientific reasons not to do so . However, AWOs indicated that retrospective analysis of all test plans submitted to the HPV Challenge Program showed that only 35% of individual repeated - dose, reproductive and developmental toxicity tests originally p roposed were subsequently replaced by proposals for combined repeated - dose reproductive/developmental toxicity screening test s . Response . EPA does, in fact, recommend use of the Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Tox icity Screening Test (40 CFR 799.9365). However, there may be valid reasons to test a particular chemical using both 40 CFR 799.9355 (Reproduction/Developmental Toxicity Screening Test) and 40 CFR 799.9305 (Repeated Dose 28 - Day Oral Toxicity Study in Roden ts) to fill the Mammalian Toxicity - Repeated Dose/Reproduction/Developmental data needs, and is requiring in the final test rule that anyone who uses the combination of 40 CFR 799.9355 and 40 CFR 799.9305 in place of 40 CFR 799.9365 submits to EPA a ratio nale for conducting these alternate tests. Comment . AWOs stated that an embryonic stem cell test (E S T) has been validated by the ECVAM as a test for embryotoxicity (Buesen et al., 2009; Festag et al., 2007a, 2007b; Genschow et al., 2002; Spielmann, 2009; van Dartel et al., 2010) . AWOs suggested that this test is ideally suited for the HPV Challenge Program whereby chemicals testing positive for embryotoxicity could be classified as probable developmental toxicants without further testing. Response . EPA do es not agree. In a recent article on the embryonic stem cell test (EST; Buensen et al. 2009), the authors and developers of the original EST assay acknowledge that the version of the test that was validated by the ECVAM has since been shown to have some si gnificant drawbacks. The main drawbacks, from EPA’s perspective, are the test’s reliance on a morphological endpoint (contracting cardiomyocytes) and the training and validation required to ensure that personnel can reliably obtain repeat assessments for t his endpoint. The measurement for this endpoint is subjective rather than objective. In an attempt to address these issues, as well as the issue of time and expense of testing, Buensen et al. (2009) have11 established a new version of the EST, which they ref er to as molecular fluorescence activated cell sorting (FACS) - EST . However, the authors acknowledge that further studies which incorporate a much wider selection of test compounds are necessary to assess the capabilities and limitations of the new techniqu e. D. Animal Welfare Considerations Comment . AWOs expressed concern that EPA is still proposing extensive animal testing of HPV chemicals and that these proposals continue to ignore existing information and basic animal welfare principles that EPA enumer ated in its October 14, 1999 letter to program participants . AWOs stated that experience gained during the review of hundreds of test plans should be applied to reduce the need for additional animal testing . This experience was summarized in comments submi tted by the Physicians Committee for Responsible Medicine (PCRM) to EPA on June 4, 2008 regarding the Chemicals Assessment and Management Program (ChAMP) . The comments by PCRM were included as an attachment to AWO comments on the proposed test rule. Respo nse . EPA agrees with AWOs that experience gained during the review of hundreds of test plans should be applied to reduce the need for additional animal sacrifices testing . The testing in this rule was required only after extensive efforts by industry, public commenters and EPA scientists to determine the availability of extant data to satisfy the required aquatic and human health endpoints for the test rule chemicals, the av ailability of supporting chemical data that would cover the needed endpoints in lieu of the need to collect further data, the use of available in vitro data where acceptable, as well as modeled data and weight - of - evidence approaches where appropriate . All available and currently acceptable approaches combined were used to eliminate as much additional animal testing as possible . After exhausting all other available means available to satisfy the required endpoints, the Agency determined that tests involving the use of animals for these 16 chemicals were required to provide the data needed to guide risk assessments and risk management actions that would protect the environment and human health . While additional test rules for HPV chemicals may be promulgated by EPA, similar approaches as outlined above will be taken to minimize the use of live animals in testing.12 Once the required data are obtained on these chemicals, they will add to our compendium of knowledge on the toxicity of chemicals . EPA expects that this additional knowledge will aid in reducing the need for further animal testing on comparable chemicals in the future. Comment . AWOs urged OPPT to implement an approach outlined in the National Research Council’s report , “ Toxicity Testing in the Twenty - first Century: a Vision and a Strategy ” (NRC, 2007) in which a recommendation was made to shift from whole animal tests that assess apical endpoints toward more human - relevant in silico and in vitro methods that assess toxicity at the molecular level. Re sponse . EPA, the National Toxicology Program (NTP) and the National Institutes of Health Chemical Genomics Center (NCGC) have formed a partnership known as Tox21, dedicated to the advancement of toxicity testing . Tox21 has responded to the NRC report with the publication in March 2009 of The U.S. Environmental Protection Agency’s “Strategic Plan for Evaluating the Toxicity of Chemicals” ( http://www.epa.gov/osa/spc/toxicitytesting/ ). In this plan, EPA states its commitment to the use of new scientific tools in computational, information and molecular sciences to strengthen toxicity testing and risk assessment approaches. The change from using studies in animals to a reliance on results from biochemical and cell - based assays for regulatory decisions is a process requiring sufficient time to determine that predictions made by these new methods are adequate and reliable . E. Chemical Specific Comments: 1. Benzene, 1,2 - dimethyl - 3 - nitro - (3 - NOX) (CASRN 83 - 41 - 0). Comment . BASF, the only producer of this chemical substance , indicates in their comment that 3 - NOX is no longer a HPV chemical. Data from the most recent IUR cycles (2002 and 2006) show that the manufacture/import of this substance is well below the 1 million lb threshold established for HPV status. BASF noted tha t its IUR - reported production volume of 3 - NOX13 has been less than 100,000 pounds since 2002, and stated that the company has no plans to exceed 100,000 pounds in the future. BASF also submitted information indicating that the re is some additional production of 3 - NOX . However, this additional production of 3 - NOX is a s a byproduct produced during the production of another registered chemical and this byproduct is incinerated and managed according to a RCRA permit. Therefore, the chemical is not a high producti on volume chemical and is not subject to the rule. Response: BASF is the only producer of this chemical substance and EPA finds the additional information submitted by the company supportive of its claims. Therefore , EPA agrees 3 - NOX appears to no longe r meet the production volume or environmental release B finding thresholds and is not including this chemical in this final test rule. However, EPA may reevaluate this decision based on IUR data to be reported later in 2011. In the event IUR or other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for 3 - NOX, EPA will take appropriate steps to proceed with a test rule for th is chemical substance. Comment . AWOs noted that the proposed test rule requires mammalian acute toxicity and combined repeated - dose reproduction/developmental toxicity screening tests for 3 - NOX . AWOs found an abstract from a 1994 German toxicology assessment (Anonymous, 1994) in the TOXLINE database that summarized results of acute and repeated - dose oral toxicity of 3 - NOX in rats . AWOs stated that EPA should review the information in this assessment regarding acute and repeated - dose toxicity and determine whether reproductive organs were examined in the repeated dose toxicity study. Response: EPA ret rieved the 1994 German toxicology assessment for 3 - NOX (BG Chemie, 1994) that was mentioned by the AWOs . Although the information regarding acute and repeated - dose (28 - day) oral toxicity studies in rats were cited to unpublished studies from chemical compa nies in Germany, the BG Chemie (1994) summaries of these studies provided sufficient information for EPA to consider the studies to adequately characterize the acute and repeated - dose hazards of oral exposure to 3 - NOX .14 Comment . AWOs stated that data for a “similar” chemical (1,2 - dimethyl - 4 - nitrobenzene [CASRN 99 - 51 - 4]) were submitted to the HPV Program and that EPA considered the submitted data adequate for assessing health effects endpoints for 1,2 - dimethyl - 4 - nitrobenzene . AWOs noted that data for severa l analogues, including 2,4 - dimethyl - 1 - nitrobenzene (CASRN 89 - 87 - 2) and 4 - nitrotoluene (CASRN 99 - 99 - 0), were used in the HPV submission for 1,2 - dimethyl - 4 - nitrobenzene to complete the SIDS dataset and that EPA considered results of existing reproductive and developmental toxicity data for 4 - nitrotoluene to be adequate . AWOs urged EPA to consider this additional information in its assessment of 3 - NOX . AWOs consider it likely that the weight of the additional evidence is sufficient to eliminate the toxicity te sting requirements for this chemical . The toxicity testing requirements in the proposed test rule for benzene, 1,2 - dimethyl - 3 - nitro - include acute mammalian toxicity, in vitro or in vivo chromosomal damage, and a combined repeated - dose reproductive/develop mental screening test. Response . In its assessment of the HPV submission for 1,2 - dimethyl - 4 - nitrobenzene, EPA noted physical/chemical similarities between the sponsored chemical and analogues that included 2,4 - dimethyl - 1 - nitrobenzene and 4 nitrotoluene . T he determination that submitted information for the sponsored chemical and the analogs was adequate for assessing health effects was based in part on comparative information for acute toxicity and genotoxicity endpoints between sponsored and analogue chemi cals . However, in the absence of information regarding acute toxicity, repeated - dose toxicity, reproductive/developmental toxicity, or genotoxicity of benzene, 1,2 - dimethyl - 3 - nitro - , comparisons with data from analogue chemicals are not possible . Therefore , if 3 - NOX still met the finding of “substantial production,” EPA would still require these mammalian toxicity testing requirements listed in the proposed test rule. 2. 3 - Pentanone (CASRN 96 - 22 - 0) . Comment . AWOs stated that the proposed test rule requires mammalian acute toxicity and combined repeated - dose reproduction/developmental toxicity screening tests for 3 - pentanone . AWOs noted that a search of fragrance raw materials monographs identified a reproductive15 toxicity study of 3 - pentanone in mice (Carlson et al., 1975) . AWOs further noted that a structurally similar chemical, 2 - pentanone (CASRN 99 - 51 - 4) was sponsored in the HPV Challenge program and that all health effects endpoints were fulfilled for this chemical . AWOs urged EPA to consider the existing data for 2 - pentanone, and other short chain aliphatic ketones such as methyl ethyl ketone (CASRN 78 - 93 - 3) in its assessment of 3 - pentanone. Response . AWOs incorrectly stated the test rule requirements for 3 - pentanone . T he proposed test rule proposed a b acterial reverse mutation test and a reproduction/developmental toxicity screening test. For these proposed studies, EPA assessed existing data for 2 - pentanone submitted to the HPV Challenge Program. Based on similar chemical/physical properties and compar ative acute lethality data for 2 - pentanone and 3 - pentanone, EPA considers a read - across approach from 2 - pentanone to 3 - pentanone to be appropriate, and EPA has determined that the toxicity data submitted for 2 - pentanone to the HPV Challenge program are ade quate to satisfy the requirements for bacterial reverse mutation ( in vitro ) and reproduction/developmental toxicity screening testing of 3 - pentanone. Therefore, additional testing of 3 - pentanone is not necessary, and EPA has dropped this chemical from the test rule. 3. Benzene, 1 - chloro - 4 - (trifluoromethyl) - (PCBTF) (CASRN 98 - 56 - 6) . Comment . GCC in their comments provided results of a l og K ow study, an aerobic biodegradation study and an anaerobic biodegradation study conducted by Lilly Research Laboratories and submitted to EPA under a 1983 negotiated testing agreement ( NTA , 48 FR 32730), in which Occidental Chemical Corporation (Occidental) had agreed to a set of testing to evaluate the human health effects, environmental effects, chemical fate and physical/chemical properties of PCBTF . Response . EPA agrees that additional log K ow testing for PCBTF is unnecessary. Although the submitted stu dy does not appear to meet GLP requirements, the test itself appears valid and the measured log K ow value is close to the one estimated by EPI Suite 4.1 (3.7 vs. 3.6), which lends confidence to the results .16 However; the aerobic and anaerobic biodegradat ion studies were inconclusive, in part or large measure because the lab did not adequately addr ess the substance's volatility, which is substantial . Although substantial, it is not so high as to render irrelevant any concern (thus testing) relating to media other than air . This means that biodegradation testing should proceed . Since the rule specifies ready biodegradability, this endpoint should be determined using a guideline appropriate for volatile chemicals . In any case, it would be a good idea for the testing lab to run high - performance liquid chromatography ( HPLC ) analyses and confirm that the test substance has been delivered to the test vessels in the am ount specified in the guideline as a check to be sure that most of it is not lost to the atmos phere somewhere between removing it from the supply bottle and getting it into the inoculated test medium. This is what happened the first time PCBTF was teste d under the NTA . This requires considerable care and even with the most commonly used method s for volatiles, o ne has to inject the substance directly into the bottle through a leak - proof septum. Comment . Regarding the need to conduct acute toxicity studies in fish, invertebrates, and plants, GCC provided results of five ecotoxicity studies performed by Occidental and submitted to EPA under the 1983 negotiated testing agreement . These ecotoxicity studies address acute toxicity in rainbow trout (Hooker, 1979a) and bluegill sunfish (Hooker, 1979b), chronic toxicity in fathead minnows (Hooker, 1981a), acu te toxicity in Daphnia (Hooker, 1979c), and chronic toxicity in Daphnia (Hooker, 1979d) . GCC believes that additional toxicity tests in fish and Daphnia are not needed because (1) EPA stated in its 1985 Federal Register (50 FR 42216) notice that “the acute and subchronic environmental effects of [PCBTF] on aquatic species are well characterized [and] additional testing is not considered necessary”, and (2) the five studies are acceptable based on an evaluation under Section 5 of EPA’s 1999 Guidance for Assessing the Adequacy of Existing Data . SMC, GCC, and IsleChem believe that the only data gap for PCBTF is algal toxicity . SMC and IsleChem noted that even though a data gap exists, it is questionable whether a study can be justified gi ven the extremely low water solubility of PCBTF . GCC propose d that the final test rule be modified to require only algal toxicity testing (ASTM E 1218 - 04e1).17 Response . EPA has reviewed the studies provided by GCC and concluded that the requirements to con duct acute studies in fish and invertebrates are satisfied . However, EPA does not agree with the assertion of SMC and IsleChem that it is questionable whether a toxicity to aquatic plants study can be justified given the extremely low water solubility of P CBTF . If the water solubility of PCBTF was great enough to permit studies to be conducted with fish and invertebrates, it should be feasible to conduct a toxicity study in algae . Therefore, EPA maintains the requirement to conduct a toxicity study in alga e. Comment . Regarding the need to conduct a reproduction/developmental toxicity test, GCC, IsleChem and SMC provided results of a two - generation reproductive toxicity study in rats performed by Occidental prior to the NTA and submitted to EPA under the NT A (Hooker, 1981b) . GCC, IsleChem and SMC consider this study to be valid and acceptable for the HPV Chemical Challenge and OECD SIDS programs . In addition, GCC, IsleChem and SMC noted that the National Toxicology Program (NTP) currently has ongoing repeate d - dose inhalation toxicity and developm ental toxicity studies of PCBTF, such th at additional testing requirements would be duplicative of the NTP studies . Response . EPA has reviewed the two - generation study provided by GCC, IsleChem and SMC and found it adequate to satisfy the test rule requirement . EPA also agrees with the GCC, IsleChem and SMC that the two NTP studies to be conducted, i.e., a teratology inhalation study and a continuous breeding inhalation study in rats, will provide sufficient informat ion regarding reproductive/developmental endpoints to satisfy the test rule requirement . Therefore, EPA is not finalizing the reproduction/developmental toxicity screening test rule proposed for PCBTF. 4. Benzenesulfonic acid, 3 - nitro - , sodium salt (1:1) (CA SRN 127 - 68 - 4) . Comment . AWOs stated that the proposed test rule requires a reproductive/developmental toxicity screening test for benzenesulfonic acid, 3 - nitro - , sodium salt . AWOs noted that a search of TOXLINE identified an abstract from a 1996 report on a safety assessment for this chemical (Anonymous, 1996) in which it was stated that a median lethal dose in rats was not achieved at an oral dose of 5,000 mg/kg, that chronic administration of a composite hair dye18 formulation containing 2.25% of the test chemical did not cause gross or microscopic changes in dogs, and that the same hair dye formulation was not classified as a reproductive toxicant in rats or rabbits administere d the formulation in the diet at concentrations of 0.005 % or 0.02% . Response . EPA reviewed the study report (Wernick et al., 1975) of chronic, reproductive, and developmental toxicity that was summarized in the safety assessment for benzenesulfonic acid, 3 - nitro - , sodium salt (sodium m - nitrobenzenesulfonate) mentioned by AWOs . The study employed a composite hair dye formulation that included 15 separate dyes and 10 separate base substances; sodium m - nitrobenzenesulfonate constituted only 2.25% of the comp osite formulation, thus precluding the usefulness of the study for hazard characterization . The requirement for a reproduction/developmental toxicity screening test will be maintained. Comment . AWOs urged EPA to consider a read - across approach from the sp onsored HPV program chemicals benzenesulfonic acid (CASRN 98 - 11 - 3) and hydroxybenzenesulfonic acid (CASRN 1333 - 39 - 7) for reproductive/developmental toxicity endpoints. Response . Regarding AWOs’ suggestion to use a read - across approach from sponsored HPV p rogram chemicals , EPA reviewed its comments on mammalian toxicity testing plans for benzenesulfonic acid and hydroxybenzenesulfonic acid . EPA determined that mammalian tests would not likely provide meaningful systemic toxicity information for these two ch emicals, which are strong acids . However, the chemical behavior of b enzenesulfonic acid, 3 - nitro - , sodium salt in mammalian systems has not been provided by the AWOs and inadequate data are available from which to predict the behavior of b enzenesulfonic ac id, 3 - nitro - , sodium salt based on available results for benzenesulfonic acid and hydroxybenzenesulfonic acid . Therefore, EPA considers data regarding reproductive and developmental toxicity endpoints for b enzenesulfonic acid, 3 - nitro - , sodium salt to be necessary and reaffirms the requirement for a reproductive/developmental toxicity screening test for benzenesulfonic acid, 3 - nitro - , sodium salt . 5. 1 - Tetracosanol (CASRN 506 - 51 - 4); 1 - Hexacosanol (CASRN 506 - 52 - 5) .19 Comment . 1 - Tetracosanol was included in the proposed test rule based on volumes reported under a 2006 Preliminary Assessment Information Reporting Rule (PAIR) . The volume reported by the only manufacturer, SASOL, under PAIR was 1,980,944 Kg . A comment by SASOL indicated that an error was made in co nverting pounds to kilograms resulting in an overstatement of the volume. SASOL recalculated the volume for this chemical as 198,094.4 Kg. Therefore, the chemical is not a high production volume chemical and is not subject to the rule. Response: EPA ha s examined the corrected information submitted by SASOL and agrees 1 - tetracosanol is not a high production volume chemical and has not included 1 - t etracosanol in the test rule. However, EPA may reevaluate this decision based on IUR data to be reported late r in 2011. In the event IUR or other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for 1 - tetracosanol, EPA will further evaluate the information submitted by SASOL regarding the chemical/physical properties of this chemical substance, and, if necessary, EPA will take appropriate steps to proceed with a test rule for th is chemical substance. Comment . 1 - Hexacosanol was included in the proposed test rule based on volumes reported under a 2006 Preliminary Assessment Information Reporting Rule (PAIR ). The volume reported by the only manufacturer, (SASOL ) under PAIR was 974,283 Kg . A comment by SASOL indicated that an error was made in converting pounds to kilograms res ulting in an overstatement of the volume. SASOL rec alculated the volume for this chemical as 97,428.3 Kg. Therefore, the chemical is not a high production volume chemical and is not subject to the rule. Response: EPA has examined the corrected information submitted by SASOL and agrees 1 - hexacosanol is not a high production volume chemical and has not included 1 - h exacosanol in the test rule. However, EPA may reevaluate this decision based on IUR data to be reported later in 2011. In the event IUR or other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for 1 - hexacosanol, EPA will further evaluate the information20 submitted by SASOL regarding the chemical/physical properties of this chemical substance, and, if necessary, take appropriate steps to proceed with a test rule for th is chemical substance. Comment . SASOL requested that EPA remove 1 - tetracosanol and 1 - hexacosanol from the proposed test rule because SASOL believes that sufficient information is already available in order to determine or predict the health or environme ntal effects of these substances . SASOL does not believe that the proposed studies are necessary to adequately characterize the effects of 1 - tetracosanol and 1 - hexacosanol and, in some cases, are not technically feasible . SASOL stated that it imports 1 - tet racosanol and 1 - hexacosanol as components of mixtures of alcohols, not as individual substances; therefore, if testing were to be required, it should be as mixtures containing these substances rather than as the individual substances . Response . As detail ed below, based on data provided by SASOL and AWOs, EPA agrees that there is adequate information available to characterize the toxicological properties of 1 - tetracosanol and 1 - hexacosanol . EPA notes that using data available for other long chain alcohols is a valid read - across approach . This approach is supported by the OECD SIDS Initial Assessment Profile of the Long Chain Alcohols Category (April 2006), which concluded that a comparison of the SIDS endpoints for the linear alcohols C6

– C22 demonstrated a clear inverse relationship between chain length and toxicological properties . Therefore, read - across of toxicity from C6

— C22 alcohols to 1 - tetracosanol (C24) and 1 - hexacosanol (C26) is acceptable as conservative prediction of toxicity. Comment . AWOs noted that the proposed test rule requires fish and mammalian acute toxicity and combined repeated - dose reproduction/developmental toxicity screening tests for 1 - tetracosanol and 1 - hexacosanol which have already been evaluated as part of the Aluminum Alkoxides Category . AWOs stated that these chemicals should be removed from the chemical list because they have already been thoroughly evaluated as part of the Aluminum Alkoxides Category sponsored by The Soap and Detergent Association; AWOs provided a we b address for the evaluation that was submitted to the HPV Challenge program ( http://www.epa.gov/HPV/pubs/summaries/almalkct/c16706tc.htm ).21 Response . EPA reviewed the data submitted to the HPV Challenge program for the Aluminum Alkoxides Category and determined that long chain alcohols were tested as analogues for the aluminum alkoxides . The data obtained for the Long Chain Alcohols in the HPV submission for Aluminum Alkoxides Catego ry are the same as the data that SASOL discussed for Long Chain Alcohols . Responses to specific comments are addressed below. Comment . SASOL does not possess results for a chronic toxicity study in Daphnia or toxicity study in algae . However, SASOL state d that the aquatic toxicity of alcohols was extensively reviewed in the LCOH SIAR(OECD, 2006); some relevant conclusions were: (1) studies of acute effects in fish (96 - h LC 50 ) for single carbon chain lengths greater than C13 - 14 in fish showed no effects up to the limit of water solubility, (2) studies of acute effects in invertebrates (48 - to 96 - h EC 50 ) for single chain lengths greater than C13 showed that no effects were likely up to the limit of water solubility, (3) studies of effects in algae (72 - h E r C 5 0 ) for single chain lengths greater than C14 showed that no effects were likely up to the limit of water solubility, and (4) chronic studies in invertebrates (21 - d NOEC) for single chain lengths greater than C15 showed no effects expected up to the limit o f water solubility . SASOL stated that the chronic toxicity studies in Daphnia required making extensive modifications to the standard experimental protocol to account for the rapid biodegradation of the alcohols and low solubility . SASOL noted that even wi th the extensive modifications and extreme measures taken, significant degradation of the alcohols occurred . SASOL stated that based on the experiences with the LCOHs tested, it would be unlikely that a chronic Daphnia study with 1 - tetracosanol and 1 - hexac osanol would be technically possible. Response . I nformation provided by SASOL and information in EPA’s Screening - Level Hazard Characterization for Long Chai n Alcohols Category (EPA, 2009) indicate s that, for single chain lengths >C14 and >C15, respectively, no effects are likely on algal growth rate and on chronic exposure in invertebrates up to the limit of water solubility . Therefore, EPA will not require aquatic toxicity testing (i.e., chronic testing in Daphnia and toxicity to algae). Comme nt . SASOL stated that a recent review (Veenstra et al., 2009) of the health effects of the LCOH category concluded that all members had low toxicity following acute and repeated22 exposures and that they are not mutagenic, carcinogenic, or developmental and reproductive toxins . The data were found to support the overall conclusion that there is an inverse relationship between chain length and the toxicity of long chain alcohols . SASOL indicated that it is reasonable to predict that 1 - tetracosanol and 1 - hexaco sanol might have even lower toxic potency than the alcohols in the category for which information on health effects is available . According to SNA, the available data can be used as conservative predictors of the health effects of 1 - tetracosanol and 1 - hexa cosanol. Response . EPA agrees that information regarding the LCOH category (C6

– C22) can be used as a conservative predictor for the health effects of 1 - tetracosanol and 1 - hexacosanol . EPA’s Screening Level Hazard Characterization for the Long Chain Alcoho ls Category (2009), which is based upon the OECD SIAR for long chain alcohols, states that the category is justified on the basis of similarities in structural, physicochemical, environmental and health parameters . That document also concluded that a clear relationship between chain length and toxicological properties (as chain length increased, toxicity decreased) justified read across from shorter chain alcohols to category members . In addition, SASOL noted that part of the health effects database in th e LCOH SIAR (OECD, 2006) includes studies of Alcohols, C24 - 34, which is a mixture of long chain alcohols derived from beeswax. This substance is composed of a mixture of long - chain alcohols including significant amounts of 1 - tetracosanol (13.24%) and 1 - hex acosanol (15.34%). Information from the studies conducted with long chain alcohols derived from beeswax was evaluated along with the additional data provided by SASOL below. Comment . Regarding the requirement to conduct acute oral toxicity studies for 1 - tetracosanol and 1 - hexacosanol, SASOL noted that the LCOH SIAR (OECD, 2006) cites existing oral LD 50 values for linear alcohols (1 - octanol to 1 - docosanol [C22]) that range from >2,000 mg/kg to well over 10,000 mg/kg data; most values represent the maximum administered dose . SASOL believes that acute oral LD 50 s for 1 - tetracosanol and 1 - hexacosanol would fall within this range and are more likely to be even higher . SASOL indicated that conducting the proposed studies23 would not further the understanding of the low acute toxicity of these substances and would result in unnecessary use of animals for testing. Response . EPA agrees that based on information obtained for the LCOH category, it is reasonable to predict that 1 - tetracosanol and 1 - hexacosanol will have low acute toxicity . EPA’s Screening Level Hazard Characterization for the Long Chain Alcohols Category (EPA, 2009) concluded that the acute toxicity is low for the category members following oral, dermal, or inhalation exposure . These data satisf y the test rule endpoint. Therefore, EPA will not require the Mammalian Toxicity - Acute endpoint for 1 - t etracosanol and 1 - h exacosanol . Comment . Regarding the requirement to conduct a combined repeated - dose reproduction/developmental toxicity screening test, SASOL noted that the LCOH SIAR (OECD, 2006) references a number of repeated - dose studies of structurally - related long chain alcohols (a table is provided) . SASOL stated that these studies provide a sufficient basis to predict a low order of repeated - dose toxicit y for 1 - tetracosanol and 1 - hexacosanol . SASOL indicated that conducting the proposed studies would not further the understanding of the low acute toxicity of these substances and would result in unnecessary use of animals for testing. Response . EPA has ev aluated the information presented in the LCOH SIAR (OECD, 2006) and confirmed that numerous repeated duration studies have been conducted with alcohols of varying carbon chain length, including C24

– 34 (derived from beeswax) . Robust summaries of these studi es are also available as part of the Aluminum Alkoxides Category . EPA has also confirmed that the reliability score of these studies is 1 or 2 . (Note: A score of 1 equals “reliable without restrictions” and is assigned to “studies or data...generated ac cording to generally valid and/or internationally accepted testing guidelines (preferably performed according to GLP) or in which the test parameters documented are based on a specific (national) testing guideline...or in which all parameters described are closely related/comparable to a guideline method ; ” A score of 2 equals “reliable with restrictions” and is assigned to “studies or data...(mostly not performed according to GLP), in which the test parameters documented do not totally comply with the spe cific testing guideline, but are sufficient to accept the data or in which investigations are described which cannot24 be subsumed under a testing guideline, but which are nevertheless well documented and scientifically acceptable.” ) ” ( OECD, 2005) . )

EPA agrees that based on the information available, it is reasonable to conclude that the predicted repeated - dose toxicity NOAELs and LOAELs for 1 - tetracosanol and 1 - hexacosanol will fall within the range of values for the tested long chain alcohols . The r e fore , these data satisf y the test rule endpoint and EPA will not require the Mammalian Toxicity - Repeated Dose/Reproduction/Developmental endpoint for 1 - tetracosanol and 1 - hexacosanol. Comment . SASOL stated that the LCOH SIAR (OECD, 2006) includes reference to studies that found no significant effects on pregnancy rates, uterine parameters, time to pregnancy, and gestation length, and no microscopic changes in the reproductive organs of rats exposed to 1 - dodecanol (C12) and 1 - octadecanol (C18). SASOL noted that studies of reproductive parameters with other long chain alcohols (C22 and C24 - 34) in rats and/or dogs for periods of up to a year showed these substances to have low reproductive toxic potency and that conducting the proposed reproductive studies with 1 - tetracosanol and 1 - hexacosanol seems unnecessary . SASOL also noted that results of developmental toxicity studies referenced in the LCOH SIAR (OECD, 2006) indicate that long chain alcohols have no potential for developmental effects and embryotoxicity or f etotoxicity (tables are provided). Response . Based on the evaluation of the information provided by SASOL and of the robust summaries of the mentioned studies, the information available is adequate to predict that 1 - tetracosanol and 1 - hexacosanol do not e xhibit potential for developmental toxicity . This is consistent with the conclusion of EPA’s Screening Level Hazard Characterization for the Long Chain Alcohols Category (EPA, 2009) that overall, the available reproductive and developmental toxicity data f or the long chain alcohols indicate no developmental toxicity and minimal maternal toxicity with repeated exposure . The information available is found adequate to characterize the Mammalian Toxicity - Repeated Dose/Reproduction/Developmental endpoint for 1 - t etracosanol and 1 - hexacosanol and EPA will not require the Mammalian Toxicity Repeated Dose/Reproduction/Developmental endpoint for 1 - tetracosanol and 1 - hexacosanol .25 Comment . Regarding the requirement to assess the mutagenicity and clastogenicity of 1 - tet racosanol and 1 - hexacosanol, SASOL stated that the LCOH SIAR (OECD, 2006) provides pertinent data for a whole range of linear alcohols (including C22 and C24

– 34) and that the results were consistently negative (examples are presented in a table) . SASOL no ted that the category of long chain aliphatic alcohols does not contain any structural elements that are of concern for potential mutagenic activity and that there is no reason to predict that 1 - tetracosanol and 1 - hexacosanol would deviate from this behavi or . SASOL stated that conducting the proposed studies would not yield any useful information. Response . EPA has evaluated the information in the LCOH SIAR (OECD, 2006) and the pertinent robust summaries of these studies available as part of the evaluation of the Aluminum Alkoxides Category and concludes that the data available are sufficient to predict with reasonable certainty that 1 - tetracosanol and 1 - hexacosanol would not be mutagenic or clastogenic . Therefore, EPA will not require the Mammalian Toxicity

– Genotoxicity endpoint for 1 - tetracosanol and 1 - hexacosanol . 6. 2 - Prope noic acid, 2 - carboxyethyl est er (B - CEA) (CASRN 24615 - 84 - 7). Comment . The Society of Chemical Manufacturers and Affiliates (SOCMA) commented on behalf of two member companies, Bimax, Inc. and Rhodia, Inc. to qu estion EPA’s addition of B - CEA to the proposed test rule . SOCMA indicates that there are only two manufacturers of this material, Bimax, Inc and Rhodia, Inc. and the combined production is well below 1 million pounds of this material and is not likely to increase significantly. Bimax, Inc and Rhodia, Inc have subsequently submi tted corrections to their respective 2006 IUR r eports that have substantiated the claim that the combined production volume for this chemical is well below 1 million pounds. Response: EPA has examined the information on production volumes for B - CEA submitted by SOCMA (and IUR corrections submitted directly from Bimax, Inc. and Rhodia, Inc.) and agrees B - CEA i s not a high production volume chemical and has not included B - CEA in the test rule. However, EPA may reevaluate this decision based on IUR data to be reported later in 2011. In26 the event IUR or other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for B - CEA, EPA will take appropriate steps to proceed with a test rule for th is chemical substance. 7. Benzenesul fo nic acid, dimethyl - (CASRN 25321 - 41 - 9) . Comment . Nease provided a rationale for using data for surrogate chemicals to satisfy the proposed test rule for benzenesulfonic acid, dimethyl - because l imited data exist regarding the physicochemical and toxicological properties of benzenesulfo nic acid, dimethyl - . Two groups of surrogates were proposed: (1) several salts of the compound, and (2) acid forms of structurally - related aromatic sulfonic acids . The use of salts was considered justified on the grounds that in aqueous media, pH >1, aromatic sulfonic acids are expected to be highly ionized . Thus, data available for benzenesulfonic acid, dimethyl - salts for many endpoints, in particular those studies that may take place in aqueous media at or near pH 7 can be used for benzenesulfonic acid, dimethyl - . The use of data on the structurally - related aromatic sulfonic acids such as benzenesulfonic acid (0 methyl groups) and p - toluenesulfonic acid (1 methyl g roup) was considered justified on the grounds that the addition of methyl groups to the aromatic ring will have only a slight electronic effect on the ring and will also “block” substituent sites, thus reducing chemical reactivity (benzenesulfonic acid, di methyl - has 2 methyl groups attached to the ring) . Nease stated that based on the predicted differences in chemical properties among these chemicals, the use of data to fill data gaps among the group is warranted. Nease noted that t here are several isomers of this test substance , although the CAS entry does not specify position of the methyl groups. Citing a ssessment s sponsored by Nease and conducted by Toxicology Excellence for Risk Assessment ( TERA ) and the Consortium for Environmental Risk Managem ent ( CERM ) , Nease concluded that all testing requirements have been previously addressed. Nease believes different isomers, as well as structural analogues, are acceptable surrogates. The study a nalys e s utilized data for xylenesulfonic acid (2,4 - dimethylbe nzenesulfonic acid) (XSA) and its salts, due to aqueous solubility and the resulting expectation of ionization in aquous media. Nease also provided data available for p -27 toluenesulfonic acid (pTSA) and benzesulfonic acid (BSA) , citing their structural simi larity to benzenesulfonic acid, dimethyl - . Nease provided data on the p hysical properties of XSA , e stimated values from EPISuite v4.0, and measured values for the HPV test substance from MSDS provided by Arichem company. Properties of the surrogate compoun ds also provided by Nease. Response . EPA has evaluated the data provided by Nease on 2,4 - xylenesulfonic acid ( 2,4 - XSA) isomer (CASRN: 88 - 61 - 9) and other analog substances , including XSA salts. With regard to physical/chemical (p - chem ) properties, EPA finds that the analogs with fewer than two methyl groups are unsatisfactory, as are the data for XSA salts. The 2,4 - XSA isomer is a solid that is recrystallized from water, whereas the test rule chemical ( XSA: CASRN 25321 - 41 - 9) is an unsp ecified mixture of isomers and a liquid (Arichem MSDS : 95% pure). Acid salt compounds have different p - chem properties from the acid forms. For example: pure 2,4 - XSA has a melting point of 61 - 62 o C and the salt of XSA (CASRN 1300 - 72 - 7), which is a mix ture of salts of isomers, has a melting point of 27 o C . T he test rule chemical ( XSA: CASRN 25321 - 41 - 9), which is a mixture of isomers, has a melting point of < 25 o C and is expected to be a liquid. T he boiling point (or decomposition point) is still nee ded, as is the vapor pressure. (Note: the Arichem MSDS listed a flashpoint of 95 °C, so vapor pressure at an elevated temperature is sufficient to allow ignition.) Solubility in water was listed at 100% by Arichem, so additional solubility data is not need ed, but the log K ow partition coefficient should be measured and this test requirement remains in the rule. Comment . Nease also provided a number of studies on surrogate chemicals and stated that these should be adequate for the ready biodegradation test ing endpoint in the rule. Response . Regarding the test requirement for ready b iodegradability, s ome of the studies cited by Nease do not fulfill the ready biodegradability endpoint, primarily because they employ the use of a microbial inoculum that has been adapted to biodegrade the test substance. A basic principle of ready biodegradation testing is that adapted microorganisms are not used.28 However, EPA finds that two studies, one using OECD guideline 301B Modified Sturm Test on CASRN 1300 - 7 2 - 7 (Xylene sulfonic acid, sodium salt) and another study using OECD 301C Modified MITI Test on CASRN 827 - 21 - 4 (also described as Xylene sulfonic acid, sodium salt) fulfill the r eady b iodegradation endpoint. Results were 87% biodegradation using the OECD guideline 301B ( Modified Sturm Test ) , and EPA finds these results acceptable . Though there was 0% biodegradation in the OECD 301C Modified MITI test, f ailure in the OECD 301C Modified MITI test alone is not usually taken as indication that a substance is n ot degradable in the environment; this test is very stringent and chemicals that fail often biodegrade rapidly in the environment and/or are found to be inherently biodegradable. Therefore EPA agrees n o further Ready Biodegradability testing is needed. Comment: Nease also stated that the studies provided could satisfy the toxicity testing endpoints proposed by EPA. Response . EPA agrees (see specific comments below) . The justification provided by Nease for using salts of benzenesulfonic acid, dimethyl and acid forms of structurally - related aromatic sulfonic acids as read - across chemicals to fill data gaps for toxicological properties of benzenesulfonic acid, dimethyl - is appropriate. Comment . Regarding the need to collect data on acute toxicity to fish and aquatic invertebrates and on toxicity to plants (algae) , Nease stated that data are available for sodium and calcium xylenesulfonate (salts of benzenesulfonic acid, dimethyl - ) . Nease provided robust summaries of these two chemicals . Ne ase noted that the test data were measured for aqueous solutions of the substances at approximately 40

– 41% active ingredient and were corrected for 100% active ingredient (Table 3 pr ovided by Nease) . According to Nease, these data are acceptable as read ac ross. Response . As indicated above, sodium and calcium xylene sulfonate are regarded as appropriate analogues for b enzenesulfonic acid, dimethyl - for read - across purposes . The only measured value for benzenesulfonic acid, dimethyl - presented in Table 3 of Nease ’s comments29 is a 24 - hour LC 50 for acute toxicity to fish of >5 mg/L . Since the study is provided a reliability score of 4 , it cannot be used to fulfill the test rule requirement (a score of 4 equals “not assig nable” and is given to “studies or data....which do not give sufficient experimental details and which are only listed in short abstracts or secondary l iterature (books, reviews, etc., ). ” ( OECD, 2005) . Nease also presented read - across values for acute toxicity to fish, invertebrates, and algae obtained with the sodium and calcium salts of benzenesulfonic acid, dimethyl - (corrected to 100% active ingredient) and provided a link to the summaries (Appendix 6 in Nease’s Table of Contents) . Summaries of these studies are also found in Appendix 5a for the sodium salt (SIDS Dossier) and Appendix 5e (SIDS Dossier) for the calcium salt . EPA reviewed these summaries and found the studies adequately conducted, and the r esults appropriate for read - across to satisfy the test rule requirement . Therefore, EPA will not finalize the test rules for aquatic toxicity for b enzenesulfonic acid, dimethyl - . Comment . Nease noted that very limited mammalian data are available for neat benzenesulfonic acid, dimethyl - , but there are some data on products that contain 80

– 94% of the chemical . Nease stated that since exposure in the environment would be to the deprotonated form, salts (K, Na, Ca, NH 4 ) are used as a surrogate for some mammal ian toxicity endpoints . Selected data are summarized in Table 4 of Nease . Two acute toxicity LD 50 s are available, an oral LD 50 and an intraperitoneal LD 50 . The oral LD 50 of >5,000 mg/kg is included in the IUCLID data set , but few details are available . Howe ver, Nease noted that this value is supported by reliable LD 50 s for the salts (summaries presented in a SIDS Dossier) . Response . The lack of detail (i.e., number of animals, vehicle, and duration of the observation period) in the study with benzenesulfoni c acid, dimethyl - summarized in the IUCLID dataset precludes its use to satisfy the endpoint of Mammalian Toxicity

– Acute . However, acute studies with the calcium and sodium salts summarized in SIDS Dossiers are adequate to satisfy the test rule requirement . Therefore, EPA will not finalize the test rule for the mammalian acute toxicity endpoint for b enzenesulfonic acid, dimethyl - . Comment . Nease stated that there are no data regarding genetic toxicity for benzenesulfonic acid, dimethyl - , but data are available for the sodium and calcium salt s (summaries presented in30 a SIDS Dossier) and for p - toluenesulfonic acid and benzenesulfonic acid (summaries presented in IUCLID datasets) . Assays for gene mutation and chromosomal abe rration were mostly negative. Response . EPA reviewed the summaries of in vitro mutagenicity and clastogenicity tests conducted with the sodium salt, in vitro mutagenicity and in vivo clastogenicity tests conducted with the calcium salt, in vitro mutagenic ity and clastogenicity tests with p - toluenesulfonic acid, and an in vitro mutagenicity test with benzenesulfonic acid and concludes that the studies were conducted appropriately and as such, can be used to fill the data gap for the test rule requirement . T herefore, EPA will not finalize the test rule for Mammalian Toxicity

– Genotoxicity for benzenesulfonic acid, dimethyl - . Comment . Nease noted that there are no data regarding the repeated - dose toxicity endpoint for benzenesulfonic acid dimethyl - , but that data on the various salts could be used as surrogates (shown in Table 4 and summaries presented in a SIDS Dossier). Response . Repeated - dose studies conducted with the sodium salt of benzenesulfonic acid dimethyl - listed in Table 4 of Nease include 90 - day dermal studies in rats and mice, two - year dermal studies in rats and mice, two 90 - day dietary studies in rats, and a 90 - day dietary study in mice . EPA has reviewed the summaries of these studies in the SIDS Dossier and concludes that they provide adequate information to satisfy the test rule requirement . Therefore, EPA will not finalize the test rule for Mammalian Toxicity

– Repeated Dose for benzenesulfonic acid, dimethyl - . Comment . Nease state d that there are no data regarding the Reproductive/Developmental Toxicity endpoint for benzenesulfonic acid dimethyl - , but there are relevant data for the salts . Nease note d that a 90 - day oral feeding study with the sodium salt in rats included an examinat ion of the reproductive organs and no effects were reported . In addition, a gestational exposure study with the calcium salt in rats reported no treatment - related effects (summaries presented in a SIDS Dossier).31 Response . The gestational exposure study wi th the calcium salt of benzenesulfonic acid dimethyl - and the 90 - day dietary studies in rats with the sodium salt of benzenesulfonic acid dimethyl - that conducted histological examinations on the reproductive organs of males and females satisfy the tests r ule requirement. Therefore, EPA will not finalize the test rule for Mammalian Toxicity

– Reproduction/Developmental for benzenesulfonic acid, dimethyl - . 8. Methanesulfonamide, N - [2 - [(4 - amino - 3 - methylphenyl)ethylamino]ethyl] -, sulfate (2:3) (CD3) (CASRN 25646 - 71 - 3) . Comment . Eastman and I3A stated that CD3 is no longer a high production volume chemical. Eastman stated that they have “ceased all manufacture and import of CD3 in 2006 and have no plans to restart...at this time." Eastman previously reported CD3 man ufacture under 2006 IUR. I3A stated that CD3 is a material used to process images that have been captured by a photosensitive silver halide

– i.e., traditional photographic film . I3A commented that the volume of CD 3 that is currently imported has decreased to an annual amount of 435,851 lbs based on 2009 production/import data . None of their member companies have produced CD3 since 2006. I3A indicates import volumes are well below the threshold, and are continually declining. The use of photographic film ha s been nearly completely replaced by digital photography . They add that there is no chance that this trend will be reversed, or that the related use of CD3 will increase. Response: EPA has reviewed the information provided by Eastman and I3A updating the past production and importation profile to the present time for CD3. EPA agrees that this substance is no longer a high production volume chemical and has not included CD3 in the test rule. However, EPA may reevaluate this decision based on IUR data to be reported later in 2011. In the event IUR or other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for CD3, EPA will take appropriate steps to proceed with a test ru le for th is chemical substance. Comment . I3A stated that the proposed test rule for CD3 requires screening level data for physical/chemical properties, environmental fate and pathways, aquatic toxicity and32 mammalian toxicity . I3A believes that existing data on CD3 and a close structural analogue, N,N - bis - (2 - hydroxyethyl) - p - phenylenediamine (CASRN 54381 - 16 - 7), provide the required information and help to avoid duplicative testing . I3A provided results of studies for CD3 that inc lude physical/chemical properties endpoints (melting point, boiling point, vapor pressure, water solubility, and Log P ow ), biodegradation, aquatic toxicity endpoints (acute toxicity to fish and aquatic invertebrates, toxicity to aquatic plants), and mammal ian toxicity endpoints (repeated - dose studies) . The data were extracted from published studies and internal company reports . Data provided for the proposed analogue chemical include repeated - dose toxicity studies and developmental and reproductive toxicity studies. Response . Regarding the endpoints for melting point, log Kow, and water solubility, EPA finds the data provided acceptable and is not requiring those testing endpoints in this final rule . Regarding the boiling point and vapor pressure endpoints, as a general rule, EPA does not accept estimated data for chemical/physical properties in lieu of measured data . However, EPA has determined for this chemical that measure d data are n ot required for these endpoints . The melting point of 168 - 171 o C with decomposition at 200 o C obviates the need for a boiling point determination . In addition, the material is a high melting stable solid and has negligible vapor pressure at room temperature, so this test endpoint is also not needed. However, in regards to toxicity testing, a lthough both the analog and subject test substance are based on a p - phenylenediamine substructure, the proposed analogue, N,N - bis - (2 - hydroxyethyl) - p - phenylenediamine (CASRN 54381 - 16 - 7), contains structural features that are not shared by the test rule substance . The commenter has not addressed whether the differences in structure between the two substances (and potentially, in their expected metabolites) would influence their toxicological properties . The structural differences in questio n reside in the side chains attached to the respective amines . Specifically, the test rule substance contains one alkyl group and one sulfonamide - substituted alkyl group on the amine . In contrast, the proposed analogue contains two hydroxyethyl groups in t hese positions . The differences in the electronic character of these two different types of functional groups (the sulfonamide is electron withdrawing, while the alcohols are electron - donating) can influence how these chemicals will interact in biological systems, including enzyme active sites . In addition, the hydroxyethyl33 groups have the potential to undergo metabolic oxidation to their corresponding carboxylic acids . A potential metabolic product from this pathway is a bis(carboxymethyl)amine, which woul d have the potential to chelate ions of biological significance, such as calcium, magnesium, iron and zinc . In the absence of any discussion concerning how and why the data generated for the proposed analogue could be expected to accurately represent the t oxicity of the test rule substance, the use of the proposed analogue is not supported . With respect to the required toxicity testing, the commenter has provided a range of aquatic toxicity studies conducted using CD3 as the test substance, including two 96 - hour acute toxicity to fish studies, both performed in fathead minnow ( Primephales promelas ), a 96 - hour acute toxicity to Daphnia study , and a toxicity to aquatic plants (algae) study . However, the method section of the summary of one study indicates that the fish were tested in containers which also held snails, water fleas, flatworms, segmented worms, side swimmers , and pill bugs as part of a “multispecies screening test.” This method of testing deviates from acceptable testing methods for the acute toxicity to fish endpoint and is , therefore , not acceptable in fulfillment of the test rule for acute toxicity to fish testing . It is not clear what impact the presence of other organisms might have on the toxicity of the test substance to the fish . The o ther study, conducted as an acute, flow - through toxicity study, appears to be acceptable, with a 96 - hour LC 50 of 3.6 mg/L obtained for the test substance . Therefore, the need for acute toxicity to fish testing has been met by this study. However, the acut e toxicity to Daphnia study, has the same weakness as the first acute toxicity to fish study described above; i.e., the Daphnia were tested in the direct presence of other aquatic organisms . Also, t he acute toxicity to aquatic plants (algae) study used an exposure period of 7 days; the test protocol for the test rule requires a 96 - hour exposure . T herefore, if this substance had been included in the final rule, the chronic toxicity to Daphnia and toxicity to aquatic plants testing requirements for CD3 would still be required. Two repeated - dose toxicity studies were submitted for which the testing was performed using CD3 . Both of these tests were performed in hamsters; one was a study conducted for 27.5 months with the test substance delivered by gavage; the other was a 2 - year cancer bioassay34 with the test material delivered in feed . Although these tests could be considered adequate to cover the requirement for repeated - dose testing, the only reproductive/developmental testing results were supplied on the prop osed analogue compound . Since , as explained above, testing on the analog is not considered acceptable for filling the requirements of the test rule for CD3, if this test substance had been included in the final rule, repr oductive/developmental testing woul d still be required . Moreover, since the repeated - dose testing was performed on hamsters, it is considered preferable that the requirement for mammalian toxicity testing be performed on rats using the combined repeated dose toxicity study with the reproduc tion/developmental toxicity screening test: 40 CFR 799.9365 . In summary, if CD3 met the finding of “substantial production , ” aquatic toxicity testing would still be required for CD3 as indicated by the proposed test rule, although further acute toxicity to fish testing would not be required . Acute toxicity to Daphnia testing and acute toxicity to aquatic plants (algae) testing would also still be required . In additio n, mammalian toxicity - repeated dose/reproduction/developmental testing would still be required . If conducted, a ll of these tests should be conducted using CD3 as the test substance. 9. 1 - Propanesulfonic acid, 2 - hydroxy - 3

- (2 - propen - 1 - yloxy) - , sodium salt (1:1 ) (CASRN 5

25 5 6 - 42 - 0 ) . Comment. GE submitted comments requesting exemption from the final rule based on technical information provided to EPA that their production does not create any substantial risk to human health or the environment based on exposure. The case for inclusion in this te st rule is based on total production volume for all manufacturing sites. GE is not claiming they no longer manufacture the chemical. They are only stating that they think there is limited exposure from their manufacturing process. GE submitted comment requ esting exemption from test rule because use/processing does not result in signifcant risk to humans or environment. Response: GE contributes to a national aggregate of more than 1 million pounds. The exposure criteria (general population, consumers, workers) are also met based on aggregate35 information. Therefore, this chemical will remain in the final test rule, and GE, as a manufacturer of this substance is also subject to the final test rule. 10. Coal/Coal Tar Chemical S ubstances: Solvent naphtha (coal) (CASRN 65996 - 79 - 4); Tar oils, coal (CASRN 65996 - 82 - 9); Tar, coal, high - temperature (CASRN 65996 - 89 - 6); Distillates (coal tar) (CASRN 65996 - 92 - 1); Pitch, coal tar - petroleum (CASRN 68187 - 57 - 5); Extract residues (coal), t ar oil alk., naphthalene distn. residues (CASRN 73665 - 18 - 6) . Comment . ACCCI considers the six coal tar - derived chemicals included in the proposed test rule to represent a category of similar substances because they are all derived from coal tar and are co nsidered to share similar chemical compositions and properties . Therefore, ACCCI considers coal tar products such as coal tar creosote, coal tar, and a large number of other coal tar products (some of which are undergoing evaluation under the European Regi stration, Evaluation, Authorization and Restriction of Chemicals [REACH] program) to be analogues to the six coal tar - derived test rule substances . ACCCI proposed a read - across approach between the coal tar - derived test rule substances and the analogues to predict chemical and toxicological properties for those substances without readily available data . ACCCI submitted compositional data for the six coal tar - derived test rule substances and for coal tar creosote (one of the proposed analogues) . ACCCI includ ed a table (Table 2.4) that presents information regarding proposed analogues for the coal tar - derived test rule substances based on similarities in the distillation process, physical properties, and chemical composition . ACCCI stated that the submission o f coal tar products to the REACH program likely includes relevant information, but that time constraints of the proposed test rule deadline made it impossible to obtain this information and assess it for relevance . ACCCI stated that no further testing of c oal tar - derived test rule substances should be initiated prior to evaluation of data for coal tar substances submitted to the REACH program and evaluation of data for coal tar petroleum pitch submitted to the HPV program as Heavy Fuel Oils Category.36 Besid es the above comments submitted on May 26, 2010 , ACCCI submitted an additional set of comments on July 29, 2010. In these comments ACCI requests that the above six chemicals be removed from the test rule. This request is based on the results of an independent survey commissioned by ACCI to provide accurate and reliable estimates of current exposure to the six coal - derived chemicals. The survey concludes that there is not adequate support for a finding of significant or substantial exposure for any of the six substances. The commissioned report concludes that the discrep ancy between information on consumer and commercial use contained in EPA’s Exposure Findings Document and the commissioned survey results appear to be due in part, to mistakes in reporting that may have been reflected in the 2006 IUR reports filed by certa in facilities. Response: EPA read and evaluated the comments and all substantive information submitted by AC C CI including the independent commissioned survey of all manufacturers and processors of the six coal - derived chemicals . EPA also asked for additio nal supporting information from the individual companies. The companies provided this information. Based on the confirmatory information provided by the companies, EPA agrees that these substances no longer meet the production (tar, coal, high temperature) or exposure - based findings (all six coal - tar derived chemical substances) for requiring testing under TSCA section 4(a)(1)(B). Therefore, EPA will not include these chemicals in the test rule. However, EPA may reevaluate this decision based on IUR data t o be reported later in 2011. In the event IUR or other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for any of these six coal - tar derived chemical substances, EPA will take appropriate steps to proceed with a test rule for the chemical substance(s). EPA notes that the analogues proposed for this submission appear to have been chosen on the basis of process similarities with the six test rule substances. While it is reasonable to expect that similar process will produce similar substances, the only process information available consists of the CAS definitions associated with the CAS registry numbers for the test rule substances and the proposed analogues . These definitions are broadly written and often do not prov ide enough information to assess the similarities of these streams . The suitability of any proposed analogue must be verified by comparing its compositional similarity with its37 corresponding target stream . Although it is understood that coal tar distillate s and derivatives are highly variable, complex mixtures, nearly half of the constituents of two of the six test rule substances (coal tar, high temperature and coal tar distillates) are unidentified, and nearly 88% of the constituents of a third substance (coal tar - petroleum pitch) are unidentified . Minimal compositional information is provided for the proposed analogues, so comparison to the test rule substances is impossible. In the absence of compositional data to allow direct comparison of these streams, similarities in measured data, such as physical properties, may help support a claim that two substances can possess similar characteristics . However, there are differences in physical properties (notably boiling range and vapor pressure) between some of the test rule substances and their corresponding proposed analogues that appear to signal significant differences in chemical composition . Therefore, the use of the analogues proposed by ACCCI is inadequately supported in the submission due to poor characterization of the streams involved and differences in their reported physical properties . EPA does , however, agree with the commenters with respect to not needing additional testing for ready biodegradability. Although r eady biodegradability data for the 6 subject substances (including 73665 - 18 - 6) was not provided , t he commenter did, however, provide ready biodegradability data for 5 individual components and inherent biodegradability data for one individual compone nt of the subject substances from the Japanese MITI biodegradation and accumulation database. EPA typically judge s studies from this database to be acceptable . These data indicate the majority of the components are not readily biodegradable . It follows th at mixtures of these substances or mixtures containing these substances as a major component are not likely to be readily biodegradable. The following statement is made by ACCI on page 16 of their comment document:

“ Based on limited standard ready biodegradation data (Table 2.6) and the r elatively limited solubility of the coal tar substances (Table 2.5), it is expected that biodegradation of these substances would be minimal, and it is unlikely that further testing would alter this conclusion. ”38 This statement acknowledges that the subst ances can be classified as not readily biodegradable . EPA agree s with this statement , does not think additional ready biodegradability testing is necessary , and is therefore dropping that test endpoint for these six substances . Specific comments regarding the six test rule streams and their corresponding proposed analogues are detailed below. Solvent naphtha (coal), CASRN 65996 - 79 - 4 : Proposed analogue : Distillates (coal tar), light oils, CASRN 84650 - 03 - 3 Solvent naphtha is one of the better cha racterized of the streams (92.3% of the constituents are identified in Table 2.1), with indene identified as the major component (58.5%) . The composition of the proposed analogue stream is not provided. The boiling point ranges reported in the CAS definiti ons for these two streams and reported in Table 2.4 appear to be comparable . However, the boiling ranges reported in Table 2.5 are significantly different: 83 - 87 °C for solvent naphtha and 230 - 500 °C for coal tar distillates light oils . This difference sug gests that the vapor pressure of solvent naphtha could be several orders of magnitude higher than that reported for the proposed analogue (0.005 torr). Tar oils, coal, CASRN 65996 - 82 - 9 : Proposed analogue : Distillates (coal tar), light oils, CASRN 84650 - 03 - 3 83.9% of the constituents of coal tar oils are identified in Table 2.1, with the major component identified as naphthalene (45.94%) . The composition of the proposed analogue stream is not provided. The boiling point ranges reported in the CAS de finitions for these two streams and reported in Table 2.4 appear to be comparable . However, the boiling ranges, vapor pressure ranges and water solubilities reported for these two streams in Table 2.5 differ . Differences in physical properties are likely t o reflect compositional differences between these two streams, which weaken support for using data for the proposed analogue to characterize coal tar oils. Tar, coal, high - temp., CASRN 65996 - 89 - 6 : Proposed analogue : Tar, coal, CASRN 8007 - 45 - 239 Approximatel y 50% of the constituents in high - temp coal tar are identified, with the major component identified as naphthalene (12.29%). The composition of the proposed analogue stream is not provided . The vapor pressures reported in Table 2.5 suggest that the propose d analogue may contain more volatile components than the parent stream. Distillat es (coal tar), CASRN 65996 - 92 - 1 : Proposed analogue : Anthracene oil, CASRN 90640 - 80 - 5 Proposed analogue : Distillates (coal tar) heavy oils, CASRN 90640 - 86 - 1 Approximately 55% of the constituents of coal tar distillates are identified, of which benzo(f)quinoline is the principle constituent (12.52%) . No compositional information is provided for either of the two proposed analogue streams . The CAS registry file definition for an thracene oil identifies its primary constituents as phenanthrene, anthracene and carbazole . According to the compositional information in Table 2.1, phenanthrene comprises 3.3% of coal tar distillates, carb a zole comprises 1.83% and anthracene is absent . The available information does not sufficiently demonstrate compositional similarity among these three process streams. Pitch, coal tar - petroleum, CASRN 68187 - 57 - 5 : Proposed analogue : Pitch, petroleum, arom., CASRN 68187 - 58 - 6 Proposed analogue : Pitch, co al tar, high - temp., CASRN 65996 - 93 - 2 This is the least characterized of the test rule streams submitted . Only 12.1% of the constituents have been identified, with benzo(a)pyrene as the primary component at 1.27% . Compositional information is not provided f or the two proposed analogue streams . The vapor pressures reported in table 2.5 for the two analogues differ from each other by at least 4 orders of magnitude, suggesting that these streams may not be that similar to each other, and their similarity to the test rule stream is not apparent . In addition, the source(s) for this stream are not clearly described, making comparison to the proposed analogues difficult if not impossible. Extract residues (coal), tar oil alk., naphthalene distn. residues, CASRN 736 65 - 18 - 6 : Proposed analogue : Creosote, CASRN 8001 - 58 - 9 Approximately 80% of the constituents of this stream have been identified, with acenaphthene as the primary constituent (13.91%) . Partial justification for the use of this analogue is40 presented in Tabl e 2.2; however, the compositional information is presented on the basis of broad chemical categories rather than individual constituents. Furthermore, ACCCI’s read - across strategy for toxicological endpoints is not clearly explained . ACCCI presents toxici ty data for various coal tar substances and coal tar components, but does not synthesize these data into a comprehensive table which demonstrates how available data will be read across to the six test - rule substances for each required endpoint . Data for ea ch of the required endpoints are not available for each of the proposed analogues in ACCCI’s Table 2.4, yet ACCCI claims that almost all of the test - rule required endpoints can be satisfied via read - across of data from “comparable substances” (ACCCI’s Tabl e 2.19). Comment . ACCCI considers measured data for primary constituents of coal tar substances (e.g., indene or naphthalene) to be appropriate for read across to coal tar - derived test rule substances where data are not readily available. Response . In addition to the proposed analogue process streams already discussed, the submitter presented physical property and/or fate data for 11 single chemicals that have been identified in the test rule streams . In general, single chemicals do not adequately repre sent complex process streams because they do not account for interactions between different constituents within mixtures . As shown in the table below, the different test rule streams contain varying amounts of these individual chemicals . Together, the sing le chemicals comprise between 3.2 % and 72.7% of the six test rule stream components . This information suggests that data on the primary constituents of coal tar substances are not adequate for read across to coal tar - derived test rules substances. Compone nt Coal - derived process streams, CAS RNs 65996 - 79 - 4 65996 - 82 - 9 65996 - 89 - 6 65996 - 92 - 1 68187 - 57 - 5 73665 - 18 - 9 1 - Me naphthalene 0 2.20 0.80 0 0 5.30 2 - Me naphthalene 0 5.90 1.60 0.13 0 13.70 Indene 58.50 3.00 1.30 0 0 041 Acenapthene 0 5.39 0.48 0 0.04 13.91 Anthracene 0 1.81 1.63 0 0.11 3.82 Benzo(a)pyrene 0 0 0.60 0.98 0.77 0.10 Fluoranthene 0 1.21 3.23 0 1.02 2.48 Fluorene 0 0 1.70 0 0 8.70 Naphthalene 1.37 45.94 12.29 0.01 0 1.18 Phenanthrene 0 6.51 4.50 3.30 0.37 11.74 Pyrene 0 0.76 2.42 9.41 0.92 1.69 Totals 59.87 72.72 30.55 13.83 3.23 62.62 Comment . ACCCI emphasized the fact that each of the coal tar substances is the product of a process that includes variability in feedstock and distillation methods; thus each coal tar substance is inherently a category of similar but not identical mixtures . ACCCI noted that testing of a given substance would obviate the need to infer that small differences in composition do not markedly alter toxicological properties. Response . As indicated above, ACCCI did not provide adequate information documenting that the dif ferences in composition between coal tar substances are small . Therefore, it is not clear that differences in toxicological properties between coal tar substances would be small. Comment . ACCCI stated that no two coal tar substances are identical in their chemical composition, but that the constituent compounds that contribute to aquatic toxicity are generally the same, which indicates that toxicity data for the major constituents can gene rally be used in evaluating aquatic toxicity of the coal tar substances . ACCCI presented aquatic toxicity data for major coal tar creosote constituent compounds intended to be representative of the aquatic toxicity hazard of the six coal tar - derived test r ule substances. Response . No evidence was provided to suggest that toxicity data from major coal tar creosote constituent compounds would adequately represent the toxicity of the six coal tar - derived test rule substances . As indicated above, information o n the chemical composition of coal tar substances that was provided does not adequately indicate that the coal tar substances are sufficiently similar to use as analogues for one another . The log K ow and water solubility values42 for “primary components” of coal tar substances, as shown in Table 2.5 of ACCCI’s comments, vary greatly . The log K ow of individual components ranges from 2.92 to 5.97, while the log K ow of the proposed analog creosote is 1.0; because log K ow influences aquatic toxicity, the toxicity of individual components may not be representative of the toxicity of the coal tar mixtures . Water solubility of individual components ranges from insoluble to 31 mg/L, while the water solubility of the six coal tar test - rule substances ranges from insolu ble to slightly soluble; again, these differences suggest that the toxicity of individual substances will not be representative of the toxicity of the coal tar mixtures. Comment . The proposed test rule requires aquatic toxicity testing for each of the six coal tar derived test rule substances . For the acute toxicity to aquatic invertebrates endpoint, ACCCI provided information for solvent naphtha (coal) (CASRN 65996 - 79 - 4) regarding the acute toxicity to aquatic invertebrates (48 - hour EC 50 of 1 - 6 mg/L for d aphnids) and vertebrates (96 - hour LC 50 of 20 - 50 mg/L for fathead minnows) and cited the information to a European Commission document titled “Solvent naphtha (coal) (CASRN 65996 - 79 - 4): IUCLID data set” dated February 19, 2000 (European Commission, 2000) . A CCCI also provided acute aquatic invertebrate and vertebrate toxicity data for se lected coal tar substances considered to be analogues to the test rule substances, and for selected coal tar polycyclic aromatic hydrocarbons ( PAHs ) . ACCCI considered the acute toxicity to aquatic invertebrates and vertebrates endpoints to be satisfied using data from solvent naphtha, analogue data, PAH data, and a read - across approach. Response . EPA retrieved the IUCLID dataset for solvent naphtha (European Commission, 20 00), as well as available IUCLID datasets for substances considered by ACCCI to be analogues to the coal tar - derived test rule substances . For solvent naphtha, the ACCCI and IUCLID data set summaries provided insufficient information from which to assess t he adequacy of the studies that evaluated acute toxicity to aquatic invertebrates and vertebrates endpoints . The IUCLID data set (European Commission, 2000) cited Aral Aktiengesellschaft Bochum as the source of study information, but provided no additional reference information . Additional study details would facilitate a determination of study adequacy . EPA has determined that the a category approach for the six coal tar - derived test rule substances43 suggested by ACCCI is not acceptable based on composition al and physical/chemical differences between these substances . EPA has also determined that compositional and physical/chemical differences among ACCCI - suggested analogue substances and coal tar derived test rule substances precludes the usefulness of a re ad - across approach either between the test rule substances or from the proposed analogue substances and the test rule substances . If these substances had been included in the final rule, t he a cute toxicity to aqua tic invertebrates endpoint for each of the six coal tar - derived test rule substances would still be re quired . Comment . For the chronic toxicity to aquatic organisms endpoint, ACCCI cited a 21 - day fish study that reported a LOEC of 1.9 mg/L for coal tar, one of the substances that ACCCI considers t o represent an analogue to the coal tar - derived test rule substances . ACCCI also provided results from tests of selected individual PAHs in fish and aquatic invertebrates . For the toxicity to aquatic plants endpoint, ACCCI provided summary data for algae e xposed to high temp coal tar pitch and for algae , macrophytes and duckweed exposed to coal tar creosote , substances considered by ACCCI to be analogues to the coal tar - derived test rule substances . ACCCI considered the chronic toxicity to aquatic organisms and toxicity to aquatic plants endpoints satisfied based on a read - across approach. Response . EPA has determined that compositional and physical/chemical differences among ACCCI - suggested analogue substances, individual PAHs, and coal tar - derived test ru le substances precludes the usefulness of a read - across approach . If these substances had been included in the final rule, t he chronic toxicity to Daphnia and toxicity to aquatic plants testing requirements for each of the six coal tar - derived test rule su bstances would still be required. . Comment . ACCCI stated that several microcosm studies have been performed on coal tar creosote and that results of these studies can be used to supplement the data from the standard aquatic toxicity tests for which data w ere presented. Response . Data presented from microcosm studies do not satisfy the aquatic toxicity endpoints specified in the test rule . Furthermore, EPA has determined that compositional and44 physical/chemical differences among ACCCI - suggested analogue su bstances and coal tar derived test rule substances precludes the usefulness of a read - across approach. Comment . For the mammalian acute toxicity endpoint, ACCCI presented summary acute oral toxicity data for solvent naphtha ( CASRN 65996 - 79 - 4) , for coal tar creosote (CASRN 8001 - 58 - 9) and coal tar pitch (CASRN 65996 - 93 - 2) which are considered by ACCCI to be analogue substances for the coal tar - derived test rule substances , and for primary components of coal tar substances that include indene (CASR N 95 - 13 - 6), naphthalene (CASRN 91 - 20 - 3), 2 - methylnaphthalene (CASRN 91 - 57 - 6), 1 - methylnaphthalene (CASRN 90 - 12 - 0), phenanthrene (CASRN 85 - 01 - 8), pyrene (CASRN 129 - 00 - 0), fluoranthene (CASRN 206 - 44 - 0), and benzo[a]pyrene (CASRN 50 - 32 - 8) . ACCCI also presente d summary data for acute inhalation toxicity of solvent naphtha, creosote, and indene . ACCCI indicated that the provided data satisfy the acute oral toxicity endpoint based on a read - across approach among the six coal tar derived test rule substances and b etween analogue substances, primary components of coal tar substances, and the test rule substances . Response . EPA reviewed the acute oral and inhalation toxicity information provided by ACCCI . Acute toxicity data in the IUCLID dataset for solvent naphtha (European Commission, 2000) were insufficiently summarized for the purpose of assessing study adequacy . The European Commission, 2000) cited Aral Aktiengesellschaft Bochum as the source of study information, but provided no additional reference informatio n . Additional study details would facilitate a determination of study adequacy . Based on insufficient study details provided for solvent naphtha and the determination by EPA that compositional and physical/chemical differences among ACCCI - suggested analogu e substances and coal tar - derived test rule substances precludes the usefu lness of a read - across approach. If these substances had been included in the final rule, the acute oral toxicity testing requirement for each of the six coal tar derived test rule s ubstances would still be re quired . Comment . For the repeated - dose toxicity endpoint, ACCCI presented summary data from oral studies of mice administered coal tar (CASRN 8007 - 45 - 2; considered by ACCCI to be an analogue for the coal tar - derived test rule su bstances) orally for 15 days (Weyand et al., 1991),45 28 days (Culp and Beland, 1994), 185 days (Weyand et al., 1994), 260 days (Weyand et al., 1995), or 2 years (Culp et al., 1996, 1998) . ACCCI also presented summary data from an oral study of rats administ ered a substance described by the study authors as manufactured gas plant residue (a type of coal tar formed as a byproduct of coal gasification) for 5 weeks (Chadwick et al., 1995) . ACCCI identified this substance as coal tar (CASRN 0887 - 45 - 2) . ACCCI indi cated that the submitted summaries satisfy the repeated - dose toxicity endpoint for the six coal tar derived test rule substances based on a read - across approach. Response . EPA reviewed the studies that were summarized by ACCCI for the repeated - dose toxici ty endpoint . The studies were designed to assess body weight effects, carcinogenicity, and DNA adduct formation . The 185 - day oral study of Weyand et al. (1994) was the only study to report information regarding incidences of nonneoplastic lesions and the h istopathological examinations for nonneoplastic lesions were performed only on control and high - dose animals sacrificed after 95 or 185 days of treatment . EPA determined that the studies for the repeated dose toxicity endpoint that were summarized by ACCCI did not adequately assess comprehensive noncancer endpoints at multiple dose levels . Furthermore, EPA determined that compositional and physical/chemical differences among ACCCI - suggested analogue substances and coal tar - derived test rule substances precl udes the usefulness of a read - across approach . If these substances had been included in the final rule, t he repeated - dose toxicity testing requirement for the coal tar - derived test rule substances would still be re quired . Comment . The proposed test rule requires in vitr o mutagenicity testing for each of the six coal tar - derived test rule substances . ACCCI reported the results of a bacterial mutagenic ity assay for tar, coal, high - temperature (CASRN 65996 - 89 - 6; one of the coal tar - der ived test rule substances) . ACCCI also provided results of bacterial mutagenicity assays of substances considered to be analogue coal tar substances : coal tar (no CASRN supplied by ACCCI) , coal tar identified as crude tar, pitch, oils (no CASRN supplied by ACCCI) , coal tar pitch volatiles (no CASRN supplied by ACCCI) , and coal tar creosote (CASRN 8001 - 58 - 9) . In each assay, the test substance was positive for mutagenicity in the presence of metabolic activation . ACCCI indicated that the provided data satisfy the in vitro mutagenicity endpoint for the coal tar derived test rule substances based on a read - across approach.46 Response . EPA considers the requirement for in vitr o mutagenicity testing of tar, coal, high temperature (CASRN 65996 - 89 - 6 ) to be satisfied by the information provided by ACCCI for this test rule substance . EPA has determined that compositional and physical/chemical differences among ACCCI - suggested analogue substances and coal tar - derived test rule substances precludes the usefulness of a rea d - across approach to the remaining five coal - tar test rule substances , and therefore these data are not acceptable substitutes for the data that would be obtained from a test rule for these five chemical substances . Therefore , if these substances had been included in the final rule, this testing would still be required. Comment . The proposed test rule requires in vitr o or in vivo chromosomal damage testing of each of the six coal tar - derived test rule substances . ACCCI cited secondary sources (ATSDR, 2002; WHO, 2004) and three primary sources (Bender et al., 1988; Burgaz et al., 1998; Yadav and Seth, 1998) that reported positive results in assays of workers exposed to coal tar substances; the results included incre ased frequencies of chromosomal abnormalities, sister chromatid exchanges, and micronuclei . ACCCI also reported increased DNA adduct levels in mice administered coal tar or coal tar waste (Culp and Beland, 1994; Culp et al., 1966; Weyand et al. 1991, 1994) . ACCCI cited US DOE (1994) as a source for reported coal tar - induced increased frequency of micronuclei in mammalian V79 cells . ACCCI noted the absence of genotoxicity data for solvent naphtha, but indicated that the primary component (indene) was negativ e for mutagenicity in an Ames test using S. typhimurium strains TA98, TA100, TA1535, and TA1537 (Curvall et al., 1982) . ACCCI stated that based on (1) the current classification of coal tar and some PAHs as carcinogens and (2) numerous available in vitro a nd in vivo assays, additional genotoxicity assays are not warranted for the coal tar - derived test rule substances, with the possible exception of solvent naphtha. Response . Chromosomal damage reported in the workers exposed to coal tar substances was not directly attributable to the coal tar substances since exposure to other potentially genotoxic chemicals occurred in these workers . The increased frequency of micronuclei in V79 cells was induced by a substance described by the study authors as low - tempera ture coal tar, which is not one of the six coal tar - derived test rule substances . EPA determined that compositional and47 physical/chemical differences among ACCCI - suggested analogue substances and coal tar derived test rule substances precludes the usefulne ss of a read - across approach . If these substances had been included in the final rule, t he chromosomal damage testing requirement for the six coal tar - derived test rule substances would still be re quired . Comment . The proposed test rule requires repeated - dose and reproduction/developmental toxicity screening tests for s olvent naphtha (coal) (CASRN 65996 - 79 - 4); t ar oils, coal (CASRN 65996 - 82 - 9); t ar, coal, high - temperature (CASRN 65996 - 89 - 6); p itch, coal tar - petroleum (CASRN 68187 - 57 - 5); and e xtract residue s (coal), tar oil alk., naphthalene distn. residues (CASRN 73665 - 18 - 6) . The proposed test rule also requires a reproduction/developmental toxicity screening test for d istillates (coal tar) (CASRN 65996 - 92 - 1) . ACCCI presented summary data from studies of rats exposed to coal tar liquid during gestation (Hackett et al., 1984; Springer et al., 1986) . ACCCI stated that the composition of the coal tar liquid samples used in these studies was “relatively similar to coal tar distillates (CASRN 65996 - 92 - 1), extra ct residues (CASRN 73665 - 18 - 9), and high temperature coal tar (CASRN 65996 - 89 - 6) . ACCCI also provided summaries of studies in which rats or rabbits were exposed to coal tar creosote (CASRN 8001 - 58 - 9) during gestation (York, 1994, 1995a, 1995b) and a study in which rats were exposed to coal tar creosote for two generations (Marcinowski, 1993) . ACCCI indicated that these data preclude the need for additional repeated - dose and reproduction/developmental toxicity testing of the six coal tar - derived test rule su bstances based on a read - across approach. Response . EPA determined that compositional and physical/chemical differences among ACCCI - suggested analogue substances and coal tar - derived test rule substances precludes the usefulness of a read - across approach . If these substances had been included in the final rule , repeated - dose and reproduction/developmental toxicity testing for the coal tar - derived test rule substances would still be re quired . Comment . ACCCI stated that, should any further testing for repea ted - dose and reproductive/developmental toxicity endpoints be required, it should be conducted in a stepwise manner . As an example, ACCCI indicated that should a reproduction/developmental toxicity screening test for coal tar distillates (one of the six co al tar - derived test rule substances) be48 conducted and that results similar to those from existing studies of coal tar and creosote would strengthen the proposed read - across approach . ACCCI also indicated that if EPA were to deem an additional combined repe ated - dose reproduction/developmental toxicity screening test necessary, such a study should be performed using solvent naphtha (one of the coal tar - derived test rule substances) due to its differences in composition from the other coal tar - derived test rul e substances. Response . As noted above, EPA determined that compositional and physical/chemical differences among ACCCI - suggested analogue substances and coal tar - derived test rule substances precludes the usefulness of a read - across approach. The potenti al acceptability of a “stepwise” scheme for testing these chemical substances would need more evidence to support their alleged similarity than was provided by ACCCI in their comments . Given additional supporting data, EPA would be willing to consider such an approach. 11. 1,4 - Benzenedicarboxylic acid, 1,4 - dimethyl ester, manuf. of, by - products from (CASRN 68988 - 22 - 7). Comment: INVISTA submitted comments claiming minimal potential for release or exposure to 1,4 - Benzenedicarboxylic acid, 1,4 - dimethyl ester, manuf. of, by - products from , and on that basis no longer meets the exposure - based finding for requiring testing under section 4. Response . EPA has reviewed the information provided by INVISTA and agrees that this substance no longer meets the environmenta l release or human exposure B - finding thresholds . Therefore, EPA is not including this chemical in the final test rule. However, EPA may reevaluate this decision based on IUR data to be reported later in 2011. In the event IUR or other data provides new or additional support for the TSCA section 4(a)(1)(B)(i) finding for this chemical substance, EPA will take appropriate steps to proceed with a test rule for 1,4 - Benzenedicarboxylic acid, 1,4 - dimethyl ester, manuf. of, by - products from . Comment: INVISTA a lso noted that it has completed 10 of the 12 studies identified in the proposed test rule and submitted final and draft reports to EPA on December 3, 2010 .49 INVISTA also stated that it has not undertaken two of the proposed studies (chronic toxicity to Daphnia and combined repeated dose toxicity study) because it believes they are no longer necessary for purposes of characterizing the substance . INVISTA stated that it has already conducted an acute toxicity test to Daphnia using the OECD 202 method . INVI STA believes the results of this study serve as a screening level ecotoxicological evaluation similar to that performed for many other HPV chemicals . Thus, INVISTA contends that a chronic study on Daphnia is not necessary . INVISTA also commented that the c ombined repeated dose toxicity study is also not necessary because such studies normally would be requested for chemicals with potential widespread distribution in the environment . INVISTA indicated that DMT residue does not have the potential for widespre ad distribution in the environment. Response: The proposed testing included melting point, boiling point, vapor pressure, water solubility and log K ow . With respect to the se data submitted by INVIST A : For the melting point determination, the use of differential scanning calorimetry and thermogravimetric analysis were performed using OECD guideline 102 . After analysis of the data, it was found that this residue had a two glass transition states at - 20C a nd 36C . Once the temperature rose above 50C, decomposition was observed . EPA finds these data acceptable and is not requiring melting point . Since decomposition was observed as the temperature rose above 50C as noted, EPA is also not requiring boiling point testing . Vapor pressure was determined using thermogravimetric analysis following OECD guideline 104 appropriately. Two tests were performed to confirm the data collected and the measurement was given in both mm Hg and Pa units. EPA finds these resul ts acceptable and would, therefore, not require vapor pressure testing. Water solubility was visually determined by stirring for 48 hours and for 115 hours in double distilled water in a flask, which the study investigators say follows OECD guideline 105 . They mention visually determining the solubility but not any other more accurate analytical method . I t is not clear how the study could adequately determine water solubility without using some analytical piece of equipment. EPA does not find this study ac ceptable as described, and if 1,4 - Benzenedicarboxylic acid, 1,4 - dimethyl ester, manuf. of, by - product s50 from, met the finding of “substantial exposure,” EPA would still require that this test ing be conducted . INVISTA also provided data for the log K ow end point . As described , the study was conducted using the HPLC method which is acceptable, however, the investigators ran the HPLC in a methanol/water mix . They also ran the experiment using known standards in the same methanol/water mixture . EPA considers, f or this method to provide good data, known standards of similar structure must be used in order to calculate the log K ow of the sample via linear regression . The standards used are not that similar to the test sample in that none are benzoic acids or ester s . If this testing were to be conducted, EPA believes this test should be rerun keeping the solvent system and standards in mind and is requiring log K ow testing in the final rule . The proposed test rule also included that ready biodegradation be determined. The ready biodegradability study submitted by INVISTA was conducted using a standard method (OECD 301B, CO2 Evolution Test (Modified Sturm Test) under GLP . Th e study is acceptable and fulfills the ready biodegradability test endpoint. No furth er ready b iodegradability testing is needed. The proposed test rule requires aquatic toxicity testing of 1,4 - b enzenedicarboxylic acid, 1,4 - dimethyl ester, manuf. of, by - products from (CASRN 68988 - 22 - 7) . According to Table 3 of the proposed test rule, the test requirements are for acute toxicity to fish and aquatic invertebrates and toxicity to aquatic plants testing if the log K ow for the test substance is less than 4.2 (i.e., the test substance belongs to Test Group 1) . For a test substance with a log K o w ≥4.2 (i.e., the test substance belongs to Test Group 2), chronic toxicity to Daphnia and toxicity to plants (algae) testing are the required tests. INVISTA submitted the full study reports to EPA on December 3, 2010 . The report provided compositional information on CASRN 68988 - 22 - 7 . According to the analysis presented on page 38 of the report (Appendix 1), approximately 66.6% of the substance has been characterized . Of the 21 known components, three major constituents have been51 identified: dimethyl 1,4 - benzenedicarboxylate (CASRN 120 - 61 - 8; 17.72%), trimethyl [1,1’ - biphenyl] - 2,4’,5 - tricarboxylate (CASRN 32741 - 92 - 7; 11.68%) and 1,4 - benzenedicarboxylic acid, (4 - methoxycarbonylphenyl)methyl methyl ester (CASRN 55334 - 51 - 5; 18.19%) . The remaining components c onsist of esters of aromatic carboxylic acids, including benzene carboxylates, biphenyl carboxylates and carboxylates of polycyclic aromatics such as fluorene and dibenzo[b,d]pyran . More than 33% of the mixture remains uncharacterized. The log K ow determ ination for the test substance was made via the HPLC method (OECD Guidelines for the Testing of Chemicals no. 117) . Table 3 in the report indicates that, when analyzed at a neutral pH, 30 component peaks were detected in the test substance, of which 15 had a log K ow ≥ 4.2 . These 15 components together comprised approximately 25% of the total mixture . The water solubility of the substance was characterized as low (< 0.983 mg/L) . INVISTA also reported that acute Daphnia testing of DMT residue demonstrated tox icity, with a 48h - EC 50 determination of 3.9 mg/L . Since nearly 25% of the proposed test rule substance is comprised of chemicals having log K ow values ≥ 4.2, the substance should be tested according to the requirements for Test Group 2 . This determinatio n is further supported by a characterization of the substance as having low solubility in water, evidence of moderate toxicity in acute Daphnia testing and no compositional information on approximately one - third of the proposed test rule substance. INVIST A provided the report of an algae toxicity study on DMT residue . According to the report, this study was conducted in accordance with several guidelines, including OECD Guideline 201 (alga, growth inhibition test) and an OECD guidance document on aquatic t oxicity testing of difficult substances and mixtures . Due to the low solubility of the substance, testing was performed using a water soluble fraction; this resulted in a 72h

– EC 50 determination of 3.4 mg/L . The submitted study is considered to be adequate , thus satisfying the requirement for toxicity to plants (algae) testing under Test Group 2 . In order to satisfy the other requirement of Test Group 2, chronic toxicity to Daphnia testing would need to be conducted on 1,4 - benzenedicarboxylic acid, 1,4 - dim ethyl ester, manuf. of, by - products from . INVISTA submitted the report of an acute Daphnia study, but not a chronic Daphnia study . Therefore, if52 this substance had remained in the test rule, a chronic toxicity to Daphnia test would still be required. The proposed test rule also requires an acute mammalian toxicity test . INVISTA provided full studies for acute oral (OECD guideline 423) and dermal (OECD guideline 402) toxicity testing in rats of the proposed test rule substance . In the acute oral test, an LD 50 > 2000 mg/kg was reported . This is based on a limit test performed at 2000 mg/kg in which a single female rat (out of a total of 6 tested among two separate tests) was sacrificed moribund on day 2 following administration of test material . In the acute dermal test, an LD 50 > 2000 mg/kg was reported, based on 24 - hour dermal application of test material at 2000 mg/kg and the survival of all treated rats . EPA considers the results to adequately characterize the hazard of acute oral and dermal exposure to CASRN 68988 - 22 - 7 . Therefore, the proposed test rule requirement for acute mammalian toxicity testing is considered satisfied. The proposed test rule also requires a bacterial reverse mutation assay and in vitro or in vivo testing for chromosomal damage . I NVISTA submitted full study reports on genetic toxicity testing of CASRN 68988 - 22 - 7 . Test results from a bacterial reverse mutation assay, a mammalian cell gene mutation assay (mouse lymphoma L5178Y cells), and an in vitro chromosome aberration test of hum an peripheral blood lymphocytes were negative with and without exogenous metabolic activation . EPA considers the proposed genotoxicity test rule requirements to be satisfied. The proposed test rule also requires repeated dose/reproduction/developmental mammalian toxicity testing. No information was provided on repeated dose/reproduction/developmental mammalian toxicity testing . These are required endpoints for testing and this requirement is not predicated on the degree of potential human exposure. In summary, if 1,4 - Benzenedicarboxylic acid, 1,4 - dimethyl ester, manuf. of, by products from , had met the finding of “substantial exposure,” EPA would still require water solubility, log K ow , and chronic toxicity to Daphnia . In addition, repeated d ose/reprodu ction/developmental mammalian toxicity would also still be required . However,53 the other test endpoints listed in the proposed rule for this chemical substance have adequate data and would not have been included in a final rule . 12. 1 - Decene, sulfurized (CASRN 72162 - 15 - 3). Comment . AWOs stated that the proposed test rule requires fish and mammalian acute toxicity and combined repeated - dose reproduction/developmental toxicity screening tests for 1 - decene, sulfurized , which has already been evaluated as p art of the Alkyl Sulfide Category . AWOs noted that it became aware through a personal communication from an American Chemistry Council (ACC) employee that this chemical was dropped from the category in the final test plan revision for the Alkyl Sulfide Cat egory because the sponsoring members discontinued manufacture and import of this chemical . AWOs stated that EPA should verify the current HPV status of this chemical . AWOs noted that EPA found the original Alkyl Sulfide Category test plan (which included 1 - decene, sulfurized) to be “minimally acceptable,” that EPA believed these chemicals were unlikely to pose acute aquatic hazards due to their hydrophobicity, and that existing mammalian acute toxicity data indicated these compounds possess low acute toxici ty hazard by a variety of exposure routes . AWOs consider the proposed fish and mammalian acute toxicity tests for 1 - decene, sulfurized (CASRN 72162 - 15 - 3) to be unnecessary. Response . The particular tests for aquatic toxicity testing of 1 - decene, sulfurize d (CASRN 72162 - 15 - 3) in the proposed test rule depend on the log K ow for the chemical . For log K ow < 4.2, testing is required for acute toxicity to fish and daphnia and toxicity to aquatic plants . For log K ow ≥ 4.2, testing is required for chronic toxicity to Daphnia and toxicity to aquatic plants. EPA has reviewed the HPV file for the “alkyl sulfide” category and noted that the original test plan submitted by the CMA Association Petroleum Additives Panel Health, Environmental and Regulatory Task Group (HE RTG) March 28, 2000, comprised five chemicals, including 1 - decene, sulfurized . The test plan included no data for 1 - decene, sulfurized regarding aquatic or mammalian toxicity endpoints.54 HERTG provided results of studies for methyl propene derivative (CASR N 68511 - 50 - 2) that assessed its acute toxicity to fish, acute toxicity to invertebrates, and toxicity to aquatic plants using water accommodated fraction or water soluble fraction methodology; these endpoints were considered satisfied for 1 - decene, sulfuri zed based on read across . In comments on the original test plan submission for the proposed alkyl sulfides category, EPA stated that the aquatic toxicity studies for methyl propene derivative were inadequate because they were not performed according to the most appropriate procedures for chemicals such as the alkyl sulfides that exhibit low water solubility and that testing should be performed at or below the water solubility limit . EPA noted that such substances are not likely to pose acute aquatic hazards , but may have potential to pose chronic aquatic hazards . EPA also suggested that aquatic toxicity testing requirements could be satisfied with a 21 - day chronic daphnid test and a toxicity to aquatic plants test . EPA noted that chronic aquatic testing may not be necessary if the water solubility of the members of the proposed alkyl sulfides category (including 1 - decene, sulfurized) are sufficiently low . EPA maintains this position for the proposed test rule that requires aquatic toxicity testing for 1 - decen e, sulfurized. HERTG provided results of mammalian acute oral toxicity studies for methyl propene derivative and propanol/dodecylthio derivative (CASRN 67124 - 09 - 8); acute inhalation toxicity studies for methyl propene derivative and trimethyl pentane deri vative (CASRN 68515 - 88 - 8); and acute dermal toxicity studies for propanol/dodecylthio derivative, trimethyl pentane derivative, and C15 - C18 alkene derivative (CASRN 67762 - 55 - 4); the mammalian acute toxicity endpoint was considered satisfied for 1 - decene, s ulfurized based on read across . EPA commented that the acute toxicity data for the four proposed alkyl sulfides category members did not show qualitative or quantitative differences and that the results suggested low acute toxicity, but also noted that the sponsor did not provide informa tion regarding the mammalian acute toxicity endpoint for 1 - decene, sulfurized, and that the biological responses for the other four proposed alkyl sulfides category members appear too low to rank the proposed category members according to biological activi ty. HERTG provided results of studies of propanol/dodecylthio derivative (28 - day oral study in rats), methyl propene derivative (13 - week dermal study in rats and 21 - and 28 - day dermal55 studies in rabbits), and trimethyl pentane derivative (28 - day inhalatio n and dermal studies in rats); the repeated - dose toxicity endpoint for 1 - decene, sulfurized was considered satisfied based on read across . EPA was not convinced of the adequacy of a category approach for the repeated - dose endpoint because different routes of exposure were used across the studies and there was some suggestion of differences in toxicity among the substances tested . Therefore, EPA maintains the proposed test rule requirement for repeated - dose testing of 1 - decene, sulfurized . The specific requi rement is for a combined repeated - dose and reproductive/developmental toxicity screening test. HERTG proposed to perform a one - generation reproduction study of propanol/dodecylthio derivative (intended to be applicable for the other alkyl sulfide category members including 1 - decene, sulfurized), and if results show low reproductive/developmental toxicity, no further testing would be considered necessary . EPA noted that a revised test plan for the proposed alkyl sulfides category, submitted by HERTG on May 18, 2005, does not include 1 - decene, sulfurized . Therefore, in the absence of information to assess the reproductive/developmental toxicity of 1 - decene, sulfurized, EPA maintains its requirement for a combined repeated - dose and reproductive/developmental t oxicity screening test as stated in the proposed test rule.56 

### Bibliography Anonymous. 1994. Toxikologische Bewertung. Berufsgenossenschaft der chemischen Industrie. Heidelberg. 93: 16. Anonymous. 1996. Final report on the safety assessment of sodium m - nitrobenzenesulfonate. J Am Coll Toxicol . 15 (4): 337 - 47. ATSDR (Agency for Toxic Substances and Disease Registry). 2002. Toxicological profile for creosote (update). Atlanta, GA. http ://www.atsdr.cdc.gov/ToxProfiles/tp.asp?id=66&tid=18 Bender, M.A., Leonard, R.C., White, O. JR., Constantino, J.P. and Redmond, C.K. 1988. Chromosomal aberrations and sister - chromatid exchanges in lynphocytes from coke oven workers. Mutation Research . 206 : 11 - 16. Brannen, K.C., Panzica - Kelley, J.M., Charlap, J.H., and Augustine - Rauch, K.A. 2009. Zebrafish: A nonmammalian model of developmental toxicology. In: Developmental Toxicology, Third Edition. D.K. Hansen and B. Abbott., eds. NY: Informa Healthca re. pp. 215 - 241. Buesen, R., Genschow, E., Slawik, B., Visan, A., Sppielmann, H., Luch, A., and Seiler, A. 2009. Embryonic stem cell test remastered: Comparison between the validated EST and the new molecular FACS - EST for assessing developmental toxicity in vitro . Toxicological Sciences 108(2): 389 - 400. Burgaz, S., Erdem, O., Karahalil, B. and Karakaya, A.E. 1998. Cytogenetic biomonitoring of workers exposed to bitumen fumes. Mutation Research . 419(1 - 1): 123 - 130. Carlson, G.L., et al. 1975. Cycloalkanon es. 7. Hypocholesterolemic activity of aliphatic compounds related to 2,8 - dibenzylcycloocatanone. Journal of Medicinal Chemistry . 18: 1024.57 Chadwick, R.W., George, S.E. and Kohan, M.J. 1995. Potentiation of 2,6 - dinitrotoluene genotoxicity in Fischer 344 r ats by pretreatment with coal tar creosote. Journal of Toxicology and Environmental Health . 44: 319 - 336. Culp, S.J. and Beland, F.A. 1994. Comparison of DNA adduct formation in mice fed coal tar or benzo[a]pyrene. Carcinogenesis . 15(2): 247 - 252. Culp, S.J., Gaylor, D.W., Sheldon, W.G., Goldstein, L.S. and Beland, F.A. 1996. DNA adduct measurements in relation to small intestine and forestomach tumor incidence during the chronic feeding of coal tar or benzo[a]pyrene to mice. Polycyclic Aromatic Com pounds . 11: 161 - 168. Culp, S.J., Gaylor, D.W., Sheldon, W.G., Goldstein, L.S. and Beland, F.A. 1998. A comparison of the tumors induced by coal tar and benzo[a]pyrene in a 2 - year bioassay. Carcinogenesis . 19(1): 117 - 124. Curvall, M., Florin, I., Jansson, T. 1982. Mutagenicity of some indoles and related compounds in the Ames test. Toxicology . 23(1): 1 - 10. EPA. (U.S. Environmental Protection Agency). 1985. OPTS. 4 - Chlorobenzotrifluoride; Decision Not to Test. Federal Register (50 FR 42216, October 18, 198 5) (TSH - FRL 2637 - 1). http://www.epa.gov/opptintr/chemtest/pubs/note52.pdf EPA. 1996. OPPT. Protection of Stratospheric Ozone: Listing of Substitutes for Ozone Depleting Substances. Federal Register (61 FR 25585, May 22, 1996) (FRL - 5467 - 1). http://www.gpo.gov/fdsys/pkg/FR - 1996 - 05 - 22/pdf/96 - 12625.pdf EPA. 2006. OPPT. Health and Safety Data Reporting; Addition of Certai n Chemicals . Federal Register (71 FR 47130, August 16, 2006) (FRL - 7764 - 7) http://www.epa.gov/fedrgstr/EPA - TOX/2006/August/Day - 16/t13489.pdf58 EPA . 2008a. OPPT. Proposed Test Rul e for the Testing of Certain High Production Volume Chemicals; Second Group of Chemicals. Federal Register (73 FR 43314, July 24, 2008) (FRL 8373 - 9). EPA . 2008b. Letter from Elizabeth Resek , Office of Science Coordination and Policy, EPA, to Samuel H. Wi lson, Ph.D., NIEHS, on August 28, 2008. http://iccvam.niehs/nih.gov/methods/acutetox/transmitFeb08/EPA - Response.pdf EPA. 2008c. Reregistration Eligibility Decision ( RED) Document for Creosote (Case 0139). United States Environmental Protection Agency . Prevention, Pesticides and Toxic Substances. EPA 739 - R - 08 - 007. September 2008. http://www.epa.gov/oppsrrd 1/REDs/creosote_red.pdf EPA. 2009. Screening Level Hazard Characterization, Long Chain Alcohols Category. http://www.epa.gov/oppt/chemrtk/hpvis/h azchar/Category_Long%20Chain%20Alcohols_Dece mber2009.pdf EPA. 2010. OPPT. Proposed Test Rule for the Testing of Certain High Production Volume Chemicals; Third Group of Chemicals. Federal Register (75 FR 8575, February 25, 2010) (FRL - 8805 - 8). http://www.federalregister.gov/articles/2010/02/25/2010 - 3734/testing - of - certain - high production - volu me - chemicals - third - group - of - chemicals#p - 3 European Commission. 2000. IUCLID dataset for solvent naphtha (coal). http://ecb.jrc.ec.europa.eu/IUCLID - DataSheets/65996794.pdf European M edicines Agency. 2008. Concept paper on single dose/acute toxicity. Doc. Ref. EMEA/CHMP/SWP/302413/2008. London, 26 June 2008. http://www.emea.europa.eu/pdfs/human/swp/30241308en.pdf59 Festag, M., et al. 2007a. An in vitro embryotoxicity assay based on the disturbance of the differentiation of murine embryonic stem cells into endothelial cells. I: Establishment of the differentiation protocol. Toxicology in Vitro . 21 (8): 1619 - 1630. Fes tag, M., et al. 2007b. An in vitro embryotoxicity assay based on the disturbance of the differentiation of murine embryonic stem cells into endothelial cells. II. Testing of compounds. Toxicology in Vitro . 21 (8): 1631 - 1640. Genschow, E., Spielman, H., Sc olz, G., Seiler, A., Brown, N., Piersma, A., Brady, M., Celemann, N., Huuskonen, H., Paillard, F., et al. 2002. The ECVAM international validation study on in vitro embryotoxicity tests. Results of the definitive phase and evaluation of prediction models. Alternatives to Laboratory Animals . 30: 151 - 176. Hackett, P.L., Rommereim, D.N. and Sikov, M.R. 1984. Developmental toxicity following oral administration of a high - boiling coal liquid to pregnant rats. Journal of Applied Toxicology . 4(1): 57 - 62. Hooker. 1979a. Acute toxicity of parachlorobenzotrifluoride to the rainbow trout Salmo gairdneri Richardson. Produced by Union Carbide Environmental Services for Hooker Chemicals & Plastics Corporation. TSCA Section 4(a) submission. OTS#0508140. Hooker. 1979b. T oxicity of parachlorobenzotrifluoride (PCBTF) to the bluegill sunfish Lepomis macrochirus Rafinesque. Produced by Union Carbide Environmental Services for Hooker Chemicals & Plastics Corporation. TSCA Section 4(a) submission. OTS#0508149. Hooker. 1979c. Acute toxicity of parachlorobenzotrifluoride (PCBTF) to the water flea, Daphnia magna Straus. Produced by Union Carbide Environmental Services for Hooker Chemicals & Plastics Corporation. TSCA Section 4(a) submission. OTS#0508141.60 Hooker. 1979d. Daphnia m agna chronic study testing parachlorobenzotrifluoride (97% active ingredient). Produced by Union Carbide Environmental Services for Hooker Chemicals & Plastics Corporation. TSCA Section 4(a) submission. OTS#0508142. Hooker. 1981a. Toxicity of parachlorobe nzotrifluoride (PCBTF) to fathead minnow ( Pimephales promelas ) embryos and larvae. TSCA Section 4(a) submission. OTS#0508145. Hooker. 1981b. Gas chromatographic assay (PCBTF) and modified 90 - day gavage and reproduction study in rats (PCBTF). Produced by E lars Bioresearch Laboratories for Hooker Chemicals & Plastics Corporation. TSCA Section 4(a) submission. OTS#0508148 Hutchinson, T.H., et al. 2003. A strategy to reduce the numbers of fish used in acute ecotoxicity testing of pharmaceuticals. Environmenta l and Toxicological Chemistry . 22: 3031 - 3036. ICCVAM. 2006. Background Review Document (BRD): In vitro cytotoxicity test methods for estimating acute oral systemic toxicity. NIH Publication No. 07 - 4518. November 2006. http://iccvam.niehs.nih.gov/methods/acutetox/inv_nru_brd.htm Jerama, S., et al. 2005. A strategy to reduce the use of fish in acute ecotoxicity testing of new chemical substances notified in the European Union. Regula tory Toxicology and Pharmacology . 42: 218 - 224. Kinsner - Ovaskainen, A. et al. 2008. ECVAM ongoing activities to meet the cosmetics 2009 deadline related to acute oral toxicity. Presented at the 15 th International Congress on In Vitro Toxicology, Stockholm, Sweden: September 25 - 28, 2008. p. 191. Mackay, D. et al. 2009. The physicochemical basis of QSARs for baseline toxicity. SAR QSAR in Environmental Research. 20(3 - 4): 393 - 414.61 Marcinowski, J. 1993. Two - generation reproduction/fertility study in rats: Lab Project Number 672 - 006. Unpublished study prepared by International Research and Development Corporation. NRC. 2007 . Toxicity Testing in the 21 st Century: A Vision and a Strategy . Committee on Toxicity Testing Assessment of Environmental Agents, National Research Council . National Academies Press, Washington, DC. OECD. ( Organisation for Economic Co - operation and Development ). Manual for Investigation of HPV Chemicals. Chapter 3: Data Evaluation. December 2005 . OECD . 2006. SIDS Initial Assessment Report for SIAM 22. Long Chain Alcohols. http://www.aciscience.org/docs/Draft_SIDS_Long_Chain_Alcohols_1.pdf OECD . 2010a. Guidance document on using cytotoxicity tests to estimate starting doses for acute oral systemic toxicity tests. Series on Testing and Assessment. No. 129. OECD . 2010b. Draft guidance document on the Fish Threshold Approach (Project 2. 23). Draft Guidance and Review Documents/Monograph, January, 2010. http://www.oecd.org/dataoecd/31/33/44229192.pdf Ruetgers - Nease Chemical, Inc. 1992. Evaluation for biodegrability in the Modified Sturm Test of S odium xylenesulfonate [1300 - 72 - 7]. State College, PA. Ru etgers - Nease Chemical, Inc. 1994. CO 2 evolution test (Modified Sturm Test) with calcium xyelenesulfonate. State College, PA. Robinson, S., Delongeas. J. - L., Donald, E., et al. 2008. European pharmaceutical company initiative challenging the regulatory requirement for acute toxicity studies in pharmaceutical drug development. Regulatory Toxicology and Pharmacology. 50: 345 - 52. Spielmann, H. 2009. The way forward in reproductive/developmental toxicity testing.62 Alternatives to Laboratory Animals . 37(6):641 - 56. Springer, D.L., Hackett, P.L., Miller, R.A. and Buschborn, R.L. 1986. Lung development and postnatal survival for rats exposed in utero to a high - boiling coal liquid. Journal of Applied Toxicology . 6: 129 - 133. Stepan Company. 1993. Bio tic Degradation (Modified Sturm test) Evaluation, in an aqueous medium, of the "ultimate" biodegradability of substances: 1736 - 1A, 1736 - 1B, 1736 - 1C, 1 736 - 1D, 1736 - 1E / 16 BF 53. Northfield, IL. U.S. DOE (Department of Energy). 1994. Tox icity studies of mild gasification products. Morgantown WV: Office of Fossil Energy DOE/MC/26018 - 3730. DE94004113. http://www.osti.go v/bridge/purl.cover.jsp;jsessionid=3F002D7AC92FF0669A646BC910F60E C0?purl=/10188108 - gx4VhH/ van Dartel, D.A., Pennings, J.L., de la Fonteyne, L.J., van Herwijnen, M.H., van Delft, J.H., van Schooten, F.J., Piersma, A.H. 2010. Monitoring developmental toxic ity in the embryonic stem cell test using differential gene expression of differentiation - related genes. Toxicological Sciences . [Epub ahead of print] Veenstra, G., et al. 2009. Human health risk assessment of long chain alcohols. Ecotoxicology and Enviro nmental Safety . 72(4): 1016 - 1030. Veith, G. et al. 2009. A baseline inhalation toxicity model for narcosis in mammals. SAR QSAR in Environmental Research. 20(5 - 6): 567 - 78. Wernick, T., Lanman, B.M., and Fraux, J.L. 1975. Chronic toxicity, teratologic, an d reproduction studies with hair dyes. Toxicology and Applied Pharmacology. 32: 450 - 460. Weyand, E.H., Chen, Y. - C., Wu, Y., Koganti, A., Dunsford, H.A. and Rodriguez, L.V. 1995. Differences in the tumorigenic activity of a pure hydrocarbon and a complex m ixture following63 ingestion: Benzo[a]pyrene vs manufactured gas plant residue. Chemical Research in Toxicology . 8: 949 - 954. York, R. 1994. Developmental toxicity study in New Zealand white rabbits: Creosote P1/P13. Laboratory Project number 672 - 002. Study conducted by International Research and Development Corporation. Unpublished. York, R. 1995a. Developmental toxicity study in rats: North American P1/P13 creosote. Study conducted by Intermational Research and Development Corporation, Mattawan, MI, for th e Creosote Council II. Report No. 671 - 020. MRID#43584201. Unpublished. York, R. 1995b. Developmental toxicity study in rats: North American P2 creosote. Study conducted by Intermational Research and Development Corporation, Mattawan, MI, for the Creosote Council II. Report No. 671 - 022. MRID#43584202. Unpublished. Weyand, E.H., Wu, Y. and Patel, S. 1994. Biochemical effects of manufactured gas plant residue following ingestion by B6C3F1 mice . Journal of Toxicology and Environmental Health . 42: 89 - 107. Weyand, E.H., Wu, Y., Patel, S., Taylor, B.B. and Mauro, D.M. 1983. Biochemical effects of coal tar in mice following ingestion. In: Polynuclear Aromatic Hydrocarbons: Formation, Metabolism and Measurement. M. Cooke and A.J. Dennis (eds). Columbus OH, Rich land WA. Battelle Press. pp. 985 - 992. WHO (World Health Organization). 2004. Coal tar creosote. Consise International Chemical Assessment Document 62. pp. 149. Yadav, J.S. and Seth, N. 1998. Effect of polycyclic aromatic hydrocarbons on somatic chromosomes of coal tar workers. Cytobios . 93: 165 - 173.64