United States kS^laMIjk Environmental Protection ^J^iniiil m11 Agency EPA/690/R-12/006F Final 3-12-2012 Provisional Peer-Reviewed Toxicity Values for 3 -Nitro-4-chlorobenzotrifluoride (CASRN 121-17-5) Superfund Health Risk Technical Support Center National Center for Environmental Assessment Office of Research and Development U.S. Environmental Protection Agency Cincinnati, OH 45268 ------- AUTHORS, CONTRIBUTORS, AND REVIEWERS CHEMICAL MANAGER Carrie R. Fleming, PhD National Center for Environmental Assessment, Cincinnati, OH CONTRIBUTOR Jason C. Lambert, PhD, DABT National Center for Environmental Assessment, Cincinnati, OH DRAFT DOCUMENT PREPARED BY ICF International 9300 Lee Highway Fairfax, VA 22031 PRIMARY INTERNAL REVIEWERS Audrey Galizia, DrPH National Center for Environmental Assessment, Washington, DC Suryanarayana V. Vulimiri, BVSc, PhD, DABT National Center for Environmental Assessment, Washington, DC This document was externally peer reviewed under contract to Eastern Research Group, Inc. 110 Hartwell Avenue Lexington, MA 02421-3136 Questions regarding the contents of this document may be directed to the U.S. EPA Office of Research and Development's National Center for Environmental Assessment, Superfund Health Risk Technical Support Center (513-569-7300). l 3-Nitro-4-chlorobenzotri fluoride ------- TABLE OF CONTENTS COMMONLY USED ABBREVIATIONS iii BACKGROUND 1 DISCLAIMERS 1 QUESTIONS REGARDING PPRTVS 1 INTRODUCTION 2 REVIEW OF POTENTIALLY RELEVANT DATA (CANCER AND NONCANCER) 3 HUMAN STUDIES 6 Oral Exposures 6 Inhalation Exposures 6 ANIMAL STUDIES 6 Oral Exposures 6 Inhalation Exposures 7 OTHER DATA (SHORT-TERM TESTS, OTHER EXAMINATIONS) 8 Tests Evaluating Carcinogenicity, Genotoxicity, and/or Mutagenicity 13 Carcinogenicity (Exposures Other Than Oral or Inhalation) 13 Other Toxicity Studies (Exposures Other Than Oral or Inhalation) 13 Short-Term Studies 13 Metabolism/Toxicokinetic Studies 13 Mode of Action/Mechanistic Studies 14 Immunotoxicity 14 Neurotoxicity 14 DERIVATION 01 PROVISIONAL VALUES 15 DERIVATION OF ORAL REFERENCE DOSES 16 Derivation of Subchronic Provisional RfD (Subchronic p-RfD) 16 Derivation of Chronic Provisional RfD (Chronic p-RfD) 16 DERIVATION OF INHALATION REFERENCE CONCENTRATIONS 16 CANCER WEIGHT-OF-EVIDENCE DESCRIPTOR 16 DERIVATION OF PROVISIONAL CANCER POTENCY VALUES 16 APPENDIX A. PROVISIONAL SCREENING VALUES 17 APPENDIX B. DATA TABLES 19 APPENDIX C. BMD OUTPUTS 20 APPENDIX D. REFERENCES 21 li 3-Nitro-4-chlorobenzotri fluoride ------- COMMONLY USED ABBREVIATIONS BMC benchmark concentration BMD benchmark dose BMCL benchmark concentration lower bound 95% confidence interval BMDL benchmark dose lower bound 95% confidence interval HEC human equivalent concentration HED human equivalent dose IUR inhalation unit risk LOAEL lowest-observed-adverse-effect level LOAELadj LOAEL adjusted to continuous exposure duration LOAELhec LOAEL adjusted for dosimetric differences across species to a human NOAEL no-ob served-adverse-effect level NOAELadj NOAEL adjusted to continuous exposure duration NOAELhec NOAEL adjusted for dosimetric differences across species to a human NOEL no-ob served-effect level OSF oral slope factor p-IUR provisional inhalation unit risk p-OSF provisional oral slope factor p-RfC provisional reference concentration (inhalation) p-RfD provisional reference dose (oral) POD point of departure RfC reference concentration (inhalation) RfD reference dose (oral) UF uncertainty factor UFa animal-to-human uncertainty factor UFC composite uncertainty factor UFd database uncertainty factor UFh interhuman uncertainty factor UFl LOAEL-to-NOAEL uncertainty factor UFS subchronic-to-chronic uncertainty factor WOE weight of evidence 111 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 PROVISIONAL PEER-REVIEWED TOXICITY VALUES FOR 3-NITRO-4-CHLOROBENZOTRIFLUORIDE (CASRN 121-17-5) BACKGROUND A Provisional Peer-Reviewed Toxicity Value (PPRTV) is defined as a toxicity value derived for use in the Superfund Program. PPRTVs are derived after a review of the relevant scientific literature using established Agency guidance on human health toxicity value derivations. All PPRTV assessments receive internal review by a standing panel of National Center for Environment Assessment (NCEA) scientists and an independent external peer review by three scientific experts. The purpose of this document is to provide support for the hazard and dose-response assessment pertaining to chronic and subchronic exposures to substances of concern, to present the major conclusions reached in the hazard identification and derivation of the PPRTVs, and to characterize the overall confidence in these conclusions and toxicity values. It is not intended to be a comprehensive treatise on the chemical or toxicological nature of this substance. The PPRTV review process provides needed toxicity values in a quick turnaround timeframe while maintaining scientific quality. PPRTV assessments are updated approximately on a 5-year cycle for new data or methodologies that might impact the toxicity values or characterization of potential for adverse human health effects and are revised as appropriate. It is important to utilize the PPRTV database flittp://hhpprtv.ornl.gov) to obtain the current information available. When a final Integrated Risk Information System (IRIS) assessment is made publicly available on the Internet (www.epa.gov/iris). the respective PPRTVs are removed from the database. DISCLAIMERS The PPRTV document provides toxicity values and information about the adverse effects of the chemical and the evidence on which the value is based, including the strengths and limitations of the data. All users are advised to review the information provided in this document to ensure that the PPRTV used is appropriate for the types of exposures and circumstances at the site in question and the risk management decision that would be supported by the risk assessment. Other U.S. Environmental Protection Agency (EPA) programs or external parties who may choose to use PPRTVs are advised that Superfund resources will not generally be used to respond to challenges, if any, of PPRTVs used in a context outside of the Superfund program. QUESTIONS REGARDING PPRTVS Questions regarding the contents and appropriate use of this PPRTV assessment should be directed to the EPA Office of Research and Development's National Center for Environmental Assessment, Superfund Health Risk Technical Support Center (513-569-7300). 1 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 INTRODUCTION 3-Nitro-4-chlorobenzotrifluoride, CAS No. 121-17-5, is a substituted nitrobenzene with the chemical structure, C7H3CIF3NO2, shown in Figure 1, and Table 1 presents the physicochemical properties. 3-Nitro-4-chlorobenzotrifluoride is a chemical intermediate of the herbicide Fluorodifen (HSDB, 2003). Figure 1. 3-Nitro-4-chlorobenzotrifluoride Structure Table 1. Physicochemical Properties Table for 3-Nitro-4-chlorobenzotrifluoride (CASRN 121-17-5)3 Property (unit) Value Boiling point (°C) 222 Melting point (°C) -2 Density (g/inL at 25°C) 1.511 Vapor pressure (Pa at 25°C) ND pH (unitless) ND Solubility in water Insoluble Relative vapor density (air = 1) ND Molecular weight (g/mol) 225.55 aSource: Chemical Book (2010). ND = No data. No Reference Dose (RfD), Reference Concentration (RfC), or cancer assessment values for 3-nitro-4-chlorobenzotrifluoride are included in the IRIS database (U.S. EPA, 201 la) or on the Drinking Water Standards and Health Advisories List (U.S. EPA, 2009). No RfD or RfC values are reported in the HEAST (U.S. EPA, 201 lb). The Chemical Assessments and Related Activities (CARA) list does not include a Health and Environmental Effects Profile (HEEP) for 3-nitro-4-chlorobenzotrifluoride (U.S. EPA, 1994). The toxicity of 3-nitro- 4-chlorobenzotrifluoride has not been reviewed by the ATSDR (2011) or the World Health Organization (WHO, 2011). CalEPA (2008, 2009) has not derived toxicity values for exposure to 3-nitro-4-chlorobenzotrifluoride. No occupational exposure limits for 3-nitro- 4-chlorobenzotrifluoride have been derived by the American Conference of Governmental 2 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Industrial Hygienists (ACGIH, 2011), recommended by the National Institute of Occupational Safety and Health (NIOSH, 2010), or adopted by the Occupational Safety and Health Administration (OSHA, 2006). The HEAST (U.S. EPA, 2011) does not report a U.S. EPA (1986) cancer weight-of-evidence classification or an oral slope factor for 3-nitro-4-chlorobenzotrifluoride. The International Agency for Research on Cancer (IARC, 2011) has not reviewed the carcinogenic potential of 3-nitro-4-chlorobenzotrifluoride. 3-Nitro-4-chlorobenzotrifluoride is not included in the 12th Report on Carcinogens (NTP, 2011). CalEPA (2008) has not prepared a quantitative estimate of the carcinogenic potential of 3-nitro-4-chlorobenzotrifluoride. Literature searches were conducted on sources published from 1900 through July 26, 2011 for studies relevant to the derivation of provisional toxicity values for 3-nitro- 4-chlorobenzotrifluoride, CAS No. 121-17-5. Searches were conducted using the EPA's Health and Environmental Research Online (HERO) database of scientific literature. HERO searches the following databases: AGRICOLA; American Chemical Society; BioOne; Cochrane Library; DOE: Energy Information Administration, Information Bridge, and Energy Citations Database; EBSCO: Academic Search Complete; GeoRef Preview; GPO: Government Printing Office; Informaworld; IngentaConnect; J-STAGE: Japan Science & Technology; JSTOR: Mathematics & Statistics and Life Sciences; NSCEP/NEPIS (EPA publications available through the National Service Center for Environmental Publications [NSCEP] and National Environmental Publications Internet Site [NEPIS] database); PubMed: MEDLINE and CANCERLIT databases; SAGE; Science Direct; Scirus; Scitopia; SpringerLink; TOXNET (Toxicology Data Network): ANEUPL, CCRIS, ChemlDplus, CIS, CRISP, DART, EMIC, EPIDEM, ETICBACK, FEDRIP, GENE-TOX, HAPAB, HEEP, HMTC, HSDB, IRIS, ITER, LactMed, Multi-Database Search, NIOSH, NTIS, PESTAB, PPBIB, RISKLINE, TRI; and TSCATS; Virtual Health Library; Web of Science (searches Current Content database among others); World Health Organization; and Worldwide Science. The following databases outside of HERO were searched for toxicity values: ACGIH, AT SDR, CalEPA, EPA IRIS, EPA HEAST, EPA OW, EPA TSCATS/TSCATS2, NIOSH, NTP, OSHA, ACToR, and RTECS. REVIEW OF POTENTIALLY RELEVANT DATA (CANCER AND NONCANCER) Table 2 provides an overview of the relevant database for 3-nitro- 4-chlorobenzotrifluoride and includes all potentially relevant repeated short-term-, subchronic-, and chronic duration studies. The phrase, "statistical significance," used throughout the document, indicates ap-walue of <0.05. 3 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Table 2. Summary of Potentially Relevant Data for 3-Nitro-4-chlorobenzotrifluoride (CASRN 121-17-5) Category Number of Male/female, Strain, Species, Study Type, Study Duration Dosimetry" Critical Effects NOAEL' BMDL/ BMCLa LOAEL' Reference (Comments) Notesb Human 1. Oral (mg/kg-day)a Subchronic ND Chronic ND Developmental ND Reproductive ND Carcinogenicity ND 2. Inhalation (mg/m3)a Subchronic ND Chronic ND Developmental ND Reproductive ND Carcinogenicity ND Animal 1. Oral (mg/kg-day)a Subchronic 10/0, Sprague-Dawley rat, gavage, 28 or 38 d 0,1,10, or 100 for 28 d; or 100 for 28 d + 300 for 10 d (Adjusted) Decreased relative brain weight; increased triglycerides ND NDr 1 Bucchi et al. (1983) PS, PR Chronic ND Developmental ND Reproductive ND Carcinogenicity ND 4 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Table 2. Summary of Potentially Relevant Data for 3-Nitro-4-chlorobenzotrifluoride (CASRN 121-17-5) Category Number of Male/female, Strain, Species, Study Type, Study Duration Dosimetry3 Critical Effects NOAEL3 BMDL/ BMCL3 LOAEL3 Reference (Comments) Notesb 2. Inhalation (mg/m3)3 Subchronic ND Chronic ND Developmental ND Reproductive ND Carcinogenicity ND ""Dosimetry: NOAEL, BMDL/BMCL, and LOAEL values are converted to human equivalent dose (HED; in mg/kg-day) or human equivalent concentration (HEC; in mg/m3) units. All long-term exposure values (4 weeks and longer) are converted from a discontinuous to a continuous (daily) exposure. Values for inhalation (cancer and noncancer) and oral (cancer only) exposure are further converted to an HEC/D. Values from animal developmental studies are not adjusted to continuous exposure. bNotes: PS = Principal study, PR = Peer reviewed. ND = No data, NDr = Not determinable. 5 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 HUMAN STUDIES Oral Exposures No studies were identified. Inhalation Exposures No studies were identified. ANIMAL STUDIES Oral Exposures The effects of oral exposure to 3-nitro-4-chlorobenzotrifluoride have been evaluated in animals in one sub chronic-duration study (Bucchi et al., 1983). Subchronic-duration Studies Bucchi et al. (1983) The peer-reviewed study by Bucchi et al. (1983) is selected as the principal study for derivation of the screening subchronic p-RfD. Although the original source was written in Italian and published in a foreign journal, EPA had the article translated on February 1, 2011. Articles published in this foreign journal are peer reviewed, but there is no statement regarding GLP compliance. Male Sprague-Dawley rats (10/treatment group) were administered 0, 1, 10, or 100 mg/kg-day of 3-nitro-4-chlorobenzotrifluoride (purity not reported) daily in dietary milk via gavage for 28 days. A separate group of 10 rats were administered 100 mg/kg-day 3-nitro- 4-chlorobenzotrifluoride for 28 days followed by 300 mg/kg-day for 10 days (average daily dose of 153 mg/kg-day). There is no indication that there was a control for this group—so results from this group are of limited use. The rats were weighed daily and food and water consumption were measured every 2 days. Urine was collected from two rats per group on Days 14, 21, and 28 and analyzed for sedimentation, albumin, glucose, and acetone levels. At study termination, blood was collected for hematology (specifics not provided) and clinical chemistry (proteinaceous nitrogen, glucose, cholesterol, triglycerides, aspartate aminotransferase [AST, referred to by study authors as SGOT], alanine aminotransferase [ALT, referred to by study authors as SGPT], gamma glutamyl transferase [yGT], and bilirubin). After necropsy, the brain, heart, lungs, thymus, liver, spleen, pancreas, kidneys, adrenal glands, testes, bladder, and seminal vesicles were weighed. Although it was stated that cytochrome P-450, analine hydroxylase, and /;-nitroani sol e-(9-dernethyl ase were measured in the microsomal fraction of the liver, no results were presented for these endpoints. Statistical methods used were not reported. The data are presented only visually without error bars and are reported in text as percent changes from control. The lack of standard deviations or individual data preclude verifying the statistical results. All animals survived until the end of treatment. The study report only provides figures for endpoints that were considered significant. No treatment-related changes in body weight, food consumption, or urinalysis were observed, but these data were not provided in the study report. Animals treated with 100 mg/kg-day or 100 mg/kg-day followed by 300 mg/kg-day showed yellowing of their fur on the ventral zone. It was stated that all treated groups consumed 12-23% more water than the controls, but no specifics were provided for each dose nor was it indicated whether this increase was dose related. Changes in clinical chemistry, hematology, and organ weights are presented in Table B. 1. There was a statistically significant increase in blood glucose levels in animals that received 100 mg/kg-day 3-nitro-4-chlorobenzotrifluoride (24.5% higher than control values) and in the 100 + 300 mg/kg-day group (41.1% increase 6 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 compared to controls). There was a statistically significant increase in triglyceride levels in animals that received 1, 10, or 100 mg/kg-day (27.8, 39.8, and 34.1% increases compared to controls, respectively). In the 100 + 300 mg/kg-day group there were also significant increases in cholesterol (22.9% increase compared to controls) and ALT and yGT (51.4% increase compared to controls). Changes in hematology were sporadic but included a statistically significant decrease in erythrocytes at 10 mg/kg-day (18.5% decrease), an increase in leukocytes at 100 mg/kg-day (80.6% increase compared to controls), and a increase in hemoglobin in the 100 + 300 mg/kg-day group (6.6% increase compared to controls). At necropsy, there were overt signs of cardiac damage and marked yellow coloration of the adrenal glands in rats administered 100 mg/kg-day or 100 + 300 mg/kg-day. Although body weight was stated to have been unaffected (data not shown), several changes in the relative organ weights were reported. There was a statistically significant increase in relative liver weights at doses >10 mg/kg-day (an 11.6% increase in the 10-mg/kg-day group, 12.1% increase in the 100-mg/kg-day group, and a 22.4% increase in the 100 + 300 mg/kg-day group compared to controls). There was a statistically significant decrease in relative brain weights in the rats administered 1, 10, or 100 + 300 mg/kg-day (11.9%, 11.7%, and 24.7%, respectively compared to controls). The 100-mg/kg-day group also had a decrease in brain weight, but this change was not statistically significant and was not reported quantitatively. However, based on graphical representation of the data, the change was of a similar magnitude to that observed in the 1- and 10-mg/kg-day dose groups. Thymus weight was increased 33% in the 10 mg/kg-day group and decreased 26.6% in the 100 + 300 mg/kg-day group compared to controls. The weight of the testes decreased by 13.2% at 10 mg/kg-day and the weight of the spleen decreased by 15.1% at 100 + 300 mg/kg-day compared to controls. Although the study authors noted a tendency for increased relative pancreas weights proportional to dose, this change was not statistically significant at any dose. Histopathological results were only preliminary and were stated to show generalized vascular damage across all organs in all treatment groups, but the incidence data were not provided. No NOAEL could be determined from the data. A LOAEL of 1 mg/kg-day is available from the data based on decreased brain weight and increased triglycerides. Chronic-duration Studies No studies were identified. Developmental Studies No studies were identified. Reproductive Studies No studies were identified. Carcinogenicity Studies No studies were identified. Inhalation Exposures There is no suitable information to provide in this regard. 7 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 OTHER DATA (SHORT-TERM TESTS, OTHER EXAMINATIONS) Several in vitro studies were identified that examined 3-nitro-4-chlorobenzotrifluoride for its potential genotoxicity (Benigni et al., 1982; Haworth et al., 1983; Mazza et al., 1986). Table 3A summarizes these in vitro genotoxicity studies, and they are further discussed below. No studies were identified that investigated the genotoxic potential of 3-nitro-4-chlorobenzotrifluoride in vivo. Other studies that were identified were limited to acute toxicity tests. Table 3B summarizes these studies, and they are further discussed below. 8 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Table 3A. Summary of 3-Nitro-4-chlorobenzotrifluoride Genotoxicity Studies Endpoint Test System Dose Concentration" Resultsb Comments References Without Activation With Activation Genotoxicity studies in prokaryotic organisms Reverse mutation Salmonella typhimurium TA98, TA100, TA1535, TA1537 0.25 (iL The study authors stated that the cytotoxic effects of 3-nitro- 4-chlorobenzotri-fluoride only permitted testing at low doses. Benigni et al. (1982) Reverse mutation Salmonella typhimurium TA98, TA100, TA1535, TA1537 NR At least 5 dose concentrations were tested. A range-finding test was conducted in S. typhimurium TA100 at up to 10 mg/plate or to the limit of solubility; however, the specific dose concentrations tested were not reported. Haworth et al. (1983) Reverse mutation Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA1538 2500 (ig/plate In an accompanying test for antimicrobial activity in S. typhimurium TA100, 315 (ig/mL was observed to be minimally inhibitory. Mazza et al. (1986) SOS repair induction ND Genotoxicity studies in nonmammalian eukaryotic organisms Mutation ND Recombination induction Aspergillus nidulans strain P, heterozygous for /? - fl uo ro -p lie n v 1 a 1 a nine resistance 1.0 |iL Results were also reviewed in Crebelli (1987). Benigni et al. (1982) Chromosomal aberration ND 9 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Table 3A. Summary of 3-Nitro-4-chlorobenzotrifluoride Genotoxicity Studies Endpoint Test System Dose Concentration" Resultsb Comments References Without Activation With Activation Chromosomal malsegregation Saccaromyces cerevisiae strain 6117 2000 (ig/mL In an accompanying test for antimicrobial activity in S. cerevisiae strain 6117, 1250 (ig/mL was observed to be minimally inhibitory. Mazza et al. (1986) Mitotic arrest ND DNA Damage Rec-assay in Bacillus subtilis strains PB 1652 (trpC2 metBlO lys3) and PB 1791 (trpC2 metBlO recE4) 1000 (ig/disk NDr 5000 and 10,000 (ig/disk dose concentrations were also tested; however, complete inhibition of the growth of the tester strains was observed at these dose concentrations. In an accompanying test for antimicrobial activity in B. subtilis PB 1652, 315 (ig/mL was observed to be minimally inhibitory. Mazza et al. (1986) Genotoxicity studies in mammalian cells—in vitro Mutation ND Chromosomal aberrations ND Sister chromatid exchange (SCE) ND 10 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Table 3A. Summary of 3-Nitro-4-chlorobenzotrifluoride Genotoxicity Studies Endpoint Test System Dose Concentration" Resultsb Comments References Without Activation With Activation DNA damage Epithelial-like human cells collected from skin and muscle explants from human embryos 1.0 (iL/mL + NDr Unscheduled DNA synthesis was observed. The study authors state that the positive results did not fit a dose-response curve; 10 |iL/mL was also tested but the results were negative. Benigni et al. (1982) DNA adducts ND Genotoxicity studies in mammals—in vivo Chromosomal aberrations ND Sister chromatid exchange (SCE) ND DNA damage ND DNA adducts ND Mouse biochemical or visible specific locus test ND Dominant lethal ND Genotoxicity studies in subcellular systems DNA binding ND aLowest effective dose for positive results; highest dose tested for negative results. b+ = Positive, ± = Equivocal or weakly positive, - = Negative, T = Cytotoxicity, NA = Not applicable, ND = No data, NDr = Not determined, NR = Not reported, NR/Dr = Not reported by the study author but determined from data. 11 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Table 3B. Other 3-Nitro-4-chlorobenzotrifluoride Studies Test Materials and Methods Results Conclusions References Carcinogenicity studies (exposures other than oral or inhalation) ND Other toxicity studies (exposures other than oral or inhalation) 4 (sex not reported), New Zealand White rabbit, acute dermal toxicity (clipped/intact or abraded skin), 24-hr exposure; 316, 1000, 3160, or 10,000 mg/kg Absence of feces followed by death observed in 1/4 animals on Day 8; body weight loss in 2/4 animals; depression, labored respiration in an unspecified number of animals Numerous cyst-like structures throughout the lobes of the liver observed in 1/4 animals administered either 3160 or 10,000 mg/kg LD50 >> 10,000 mg/kg Hazleton Labs (1992) Short-term studies 5/5 (M/F), Sprague-Dawley rat, single gavage, observed for 14 d; 460, 613, 791, 1025, 1319, or 1706 mg/kg Dark red lungs, mottled liver, pale spleen, dark zone between the cortex and medulla of the kidney, body weight loss and moderate to severe autolysis observed at death in animals administered >460 mg/kg Depression, labored respiration after 1 hr in males and females administered >613 mg/kg Convulsions in 1/5 males observed 4 hr after administration of 791 mg/kg Blanched stomach observed in animals administered 1025 mg/kg Distended stomach with thin walls observed in animals administered >1319 mg/kg LD50 (females) = 1250 mg/kg (confidence limits, 984-1588 mg/kg) LD50 (males) = 1275 mg/kg (confidence limits, 931-1747 mg/kg) Hazleton Labs (1992) Metabolism/toxicokinetic ND Mode of action/ mechanistic ND Immunotoxicity ND Neurotoxicity ND ND = No data. 12 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Tests Evaluating Carcinogenicity, Genotoxicity, and/or Mutagenicity The genotoxicity of 3-nitro-4-chlorobenzotrifluoride has been studied using several prokaryotic, nonmammalian eukaryotic, and mammalian in vitro test systems (Benigni et al., 1982; Haworth et al., 1983; Mazza et al., 1986). Table 3A summarizes these genotoxicity studies. With the exception of the finding that 3-nitro-4-chlorobenzotrifluoride caused unscheduled DNA synthesis in cultured epithelial-like human cells (Benigni et al., 1982), these studies indicate that 3-nitro-4-chlorobenzotrifluoride is not mutagenic, genotoxic, or clastogenic in vitro. Benigni et al. (1982) observed unscheduled DNA synthesis at concentrations of 1.0 and 2.0 [iL/mL 3-nitro-4-chlorobenzotrifluoride, but not 10.0 [jL/mL. Studies investigating the in vivo genotoxic potential of 3-nitro-4-chlorobenzotrifluoride were not identified. Carcinogenicity (Exposures Other Than Oral or Inhalation) No studies were identified. Other Toxicity Studies (Exposures Other Than Oral or Inhalation) One unpublished, acute study was identified that examined the effects of 3-nitro-4-chlorobenzotrifluoride through routes of exposure other than oral or inhalation (Hazleton Labs, 1992). Table 3B summarizes this and other studies on 3-nitro- 4-chlorobenzotrifluoride. In an acute dermal toxicity study, Hazleton Labs (1992) exposed four New Zealand White rabbits (sex not reported, initial body weights of 2.4-3.1 kg) to 316, 1000, 3160, or 10,000 mg/kg of 3-nitro-4-chlorobenzotrifluoride for 24 hours. In two of the animals per exposure group, the skin was clipped and left intact while in the other two animals the skin was abraded prior to the application of 3-nitro-4-chlorobenzotrifluoride. Binders were applied to the exposed area and the animals were fitted with collars to avoid ingestion of the test material. Following exposure, the researchers observed the animals for up to 14 days. One animal in the 10,000-mg/kg exposure group did not produce feces and died 8 days after exposure. An LD50 of >10,000 mg/kg was assigned by the study authors based on this death. Short-term Studies In addition to the study on acute dermal toxicity, Hazleton Labs (1992) also conducted an unpublished, acute oral toxicity study in rats. Five male and five female Sprague-Dawley rats (initial body weights of 150-212 g for males and 142-182 g for females) were administered a single dose of 460, 613, 791, 1025, 1319, or 1706 mg/kg 3-nitro-4-chlorobenzotrifluoride (purity assumed to be 100% by study authors) via gavage in corn oil. There was no indication of a concurrent control group. The animals were examined for mortality and signs of any clinical effects immediately after dosing, after 1, 4, and 24 hours, and once daily thereafter for up to 14 days. Depression and labored respiration were observed at >613 mg/kg at 1 hour after dosing. At 791 mg/kg, convulsions were observed in 1/5 males. At all dose levels necropsy revealed dark red lungs, mottled liver, pale spleen, a dark zone between the cortex and medulla of the kidney, and moderate-to-severe autolysis. The raw incidence data of these findings were not provided. It was stated that some body-weight loss was observed. At a dose of 1706 mg/kg the cardiac portion of the stomach had thickened, was white in color, and had adhered to the liver in an unspecified number of animals. Other effects to the stomach included blanching at 1025 mg/kg and distention with thin walls at 1319 and 1706 mg/kg. LD50S of 1250 mg/kg for females and 1275 mg/kg for males were estimated from this study. Metabolism/Toxicokinetic Studies No studies were identified. 13 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Mode of Action/Mechanistic Studies No studies were identified. Immunotoxicity No studies were identified. Neurotoxicity No studies were identified. 14 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 DERIVATION OF PROVISIONAL VALUES Tables 4 and 5 present a summary of noncancer and cancer reference values, respectively. IRIS data are indicated in the table, if available. Table 4. Summary of Noncancer Reference Values for 3-Nitro-4-chlorobenzotrifluoride (CASRN 121-17-5) Toxicity Type (units) Species/Sex Critical Effect p-Reference Value POD Method POD UFC Principal Study Screening subchronic p-RfD (mg/kg-day) Rat/M Decreased relative brain weight and increased triglycerides 1 x 10~4 LOAEL 1 10,000a Bucchi et al. (1983) Chronic p-RfD (mg/kg-day) None None None None None None None Subchronic p-RfC (mg/m3) None None None None None None None Chronic p-RfC (mg/m3) None None None None None None None aThe maximum allowed total composite uncertainty factor is 10,000. Table 5. Summary of Cancer Reference Values for 3-Nitro-4-chlorobenzotrifluoride (CASRN 121-17-5) Toxicity Type Species/Sex Tumor Type Cancer Value Principal Study p-OSF None None None None p-IUR None None None None 15 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 DERIVATION OF ORAL REFERENCE DOSES Derivation of Subchronic Provisional RfD (Subchronic p-RfD) No subchronic p-RfD can be derived because doing so would require the application of a composite uncertainty factor of 10,000, which is greater than the maximum allowable uncertainty factor of 3000. However, a screening subchronic p-RfD is provided in Appendix A. The Bucchi et al. (1983) subchronic study in male rats is the only study available to consider for derivation of the subchronic p-RfD. Derivation of Chronic Provisional RfD (Chronic p-RfD) No chronic p-RfD can be derived because doing so would require the application of a composite uncertainty factor in excess of 10,000. DERIVATION OF INHALATION REFERENCE CONCENTRATIONS No subchronic or chronic p-RfC values can be derived because there are no human or animal inhalation studies currently available. CANCER WEIGHT-OF-EVIDENCE DESCRIPTOR Table 6 identifies the cancer weight-of-evidence descriptor for 3-nitro- 4-chl orob enzotrifluori de. Table 6. Cancer Weight of Evidence (WOE) Descriptor for 3-Nitro-4-chlorobenzotrifluoride Possible WOE Descriptor Designation Route of Entry (Oral, Inhalation, or Both) Comments "Carcinogenic to Humans " ND ND No comments "Likely to Be Carcinogenic to Humans " ND ND No comments "Suggestive Evidence of Carcinogenic Potential" ND ND No comments "Inadequate Information to Assess Carcinogenic Potential" Selected Both No carcinogenic studies are available that analyze either route of exposure. "Not Likely to Be Carcinogenic to Humans " ND ND No comments ND = No data DERIVATION OF PROVISIONAL CANCER POTENCY VALUES The lack of data on the carcinogenicity of 3-nitro-4-chlorobenzotrifluoride precludes the derivation of quantitative estimates for either oral (p-OSF) or inhalation (p-IUR) exposure. 16 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 APPENDIX A. PROVISIONAL SCREENING VALUES For the reasons noted in the main document, it is inappropriate to derive a provisional subchronic p-RfD for 3-nitro-4-chlorobenzotrifluoride. However, information is available which, although insufficient to support derivation of a provisional toxicity value under current guidelines, may be of limited use to risk assessors. In such cases, the Superfund Health Risk Technical Support Center summarizes available information in a supplemental appendix and develops a screening value. Appendices receive the same level of internal and external scientific peer review as the main document to ensure their appropriateness within the limitations detailed in the document. Users of screening toxicity values in a supplement to a PPRTV assessment should understand that there is considerably more uncertainty associated with the derivation of a supplemental screening toxicity value than for a value presented in the body of the assessment. Questions or concerns about the appropriate use of screening values should be directed to the Superfund Heath Risk Technical Support Center. DERIVATION OF SCREENING PROVISIONAL ORAL REFERENCES DOSES Derivation of Screening Subchronic Provisional RfD (Subchronic p-RfD) There is a single repeated dose study (Bucchi et al., 1983) available on 3-nitro-4-chlorobenzotrifluoride. The study by Bucchi et al. (1983) is selected as the principal study for derivation of the screening subchronic p-RfD. The critical endpoints are decreased relative brain weight (no changes in body weight were noted) and increased triglycerides in male rats. This study was published in a foreign language; however, the journal is peer reviewed and EPA had the study translated on February 1, 2011. The study does not provide a GLP compliance statement. Although numerous endpoints were examined, histopathology was only preliminary at the time of publication. The preliminary histopathology results indicate that generalized vascular damage occurred at the dose selected for POD; however, neither incidence nor severity data were provided. The study is limited in that only male rats were examined and limited data are available for review. Details are provided in the "Review of Potentially Relevant Data" section. BMD analysis is not possible with these data because the data are presented only graphically and as percent change from controls. As the only available and acceptable study, this study represents the lowest POD for developing a subchronic p-RfD. The POD in this study is a LOAEL of 1 mg/kg-day for decreased relative brain weight and increased serum triglycerides. No dosimetric adjustments were required because the doses in the principal study were administered via gavage in mg/kg-day, presumably for 7 days per week for the entire study duration. The screening subchronic p-RfD for 3-nitro-4-chlorobenzotrifluoride, based on 1 mg/kg-day in male rats, is derived as follows: Screening Subchronic p-RfD = LOAELadj ^ UF = 1 mg/kg-day10,000 = 1 x 10~4 mg/kg-day 17 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 Table A. 1 summarizes the uncertainty factors for the screening subchronic p-RfD for 3 -nitro-4-chl orob enzotrifluori de. Table A.l. Uncertainty Factors for the Screening Subchronic p-RfD of 3-Nitro-4-chlorobenzotrifluoride UF Value Justification Notes ufa 10 A UFa of 10 is applied for interspecies extrapolation to account for potential toxicokinetic and toxicodynamic differences between rats and humans. No notes ufd 10 A UFd of 10 is selected because there are no acceptable two-generation reproduction studies or developmental studies by this route. No notes UFh 10 A UFh of 10 is applied for intraspecies differences to account for potentially susceptible individuals in the absence of information on the variability of response in humans. No notes ufl 10 A UFl of 10 is applied for using a POD based on a LOAEL because a NOAEL cannot be determined from the available database. No notes UFS 1 A UFS of 1 is applied because a subchronic study was utilized. No notes UFC <3000 10,000 Composite of the five uncertainty factors. This value is a screening value. The composite UF maximum is 10,000. 18 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 APPENDIX B. DATA TABLES Table B.l. Changes in organ Weights, Clinical Chemistry, and Hematology in Rats Exposed to 3-Nitro-4-chlorobenzotrifluoride by Gavage for 28 Days Parameter Exposure Group (mg/kg-day) 1 10 100 100 + 300a Relative Organ Weights Liver NS +11.6%b +12.1% +22.4% Brain -11.9% -11.7% NS -24.7% Thymus NS +33% NS +26.6% Testes NS -13.2% NS NS Spleen NS NS NS -15.1% Clinical Chemistry Blood glucose NS NS +24.5% +41.1% Triglycerides +27.8% +39.8% +34.1% NS Cholesterol NS NS NS +22.9% ALT & yGT° NS NS NS +51.4% Hematology Erythrocyte count NS -18.5% NS NS Leukocyte count NS NS +80.6% NS Hemoglobin NS NS NS +6.6% aThis group received 100 mg/kg-day for 28 days and then 300 mg/kg-day for an additional 10 days. bData presented are percent difference from controls as reported in study (actual values and variance were not reported). These parameters were reported together by the study authors. NS = Not significantly different from controls (0 mg/kg-day in dietary milk gavage for 28 days). 19 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 APPENDIX C. BMD OUTPUTS There are no BMD outputs to report. 20 3-Nitro-4-chlorobenzotri fluoride ------- FINAL 3-12-2012 APPENDIX D. REFERENCES ACGIH (American Conference of Governmental Industrial Hygienists). (2011) Threshold limit values for chemical substances and physical agents and biological exposure indices. Cincinnati, OH. As cited in HSDB (Hazardous Substances Data Bank). Available online at http://toxnet.nlm.nih.eov/cgi-bin/sis/htmlgen7HSDB. Accessed on July 26, 2011. 625688. ATSDR (Agency for Toxic Substances and Disease Registry). (2011) Toxicological profile information sheet. U.S. Department of Health and Human Services, Public Health Service. Available online at http://www.atsdr.cdc.gov/toxprofiles/index.asp. Accessed on July 26, 2011. 595415. Benigni, R; Bignami, M; Conti, L; et al. (1982) In vitro mutational studies with trifluralin and trifluorotoluene derivatives. Ann 1st Super Sanita 18:123-126 (Italian). 670437 Bucchi, R; Gramenzi, F; Macri, A; Ricciardi, C. (1983) Subchronic oral toxicity in the rat of 3-nitro-4-chloro-alpha,alpha,alpha-trifluorotoluene (NCTT). Ann 1st Super Sanita 19(2-3):385-389. 670407. CalEPA (California Environmental Protection Agency). (2008) All OEHHA acute, 8-hour and chronic reference exposure levels (chRELs) as on December 18, 2008. Office of Environmental Health Hazard Assessment, Sacramento, CA. Available online at http://www.oehha.ca.gov/air/allrels.html. Accessed on July 26, 2011. 595416 CalEPA (California Environmental Protection Agency). (2009) OEHHA toxicity criteria database. Office of Environmental Health Hazard Assessment, Sacramento, CA. Available online at http://www.oehha.ca.gov/riskAChemicalDB/index.asp. Accessed on July 26, 2011. 595417 Chemical Book. 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(201 lb) Health effects assessment summary tables (HEAST). Prepared by the Office of Research and Development, National Center for Environmental Assessment, Cincinnati OH for the Office of Emergency and Remedial Response, Washington, DC. Available online at http://epa-heast.ornl.gov/. Accessed on July 26, 2011. 595422. WHO (World Health Organization). (2011) Online catalogs for the Environmental Health Criteria Series. Available online at http://www.who.int/ipcs/publications/ehc/en/. Accessed on July 26, 2011. 595424. 23 3-Nitro-4-chlorobenzotri fluoride ------- |