&EPA United States EPA/690/R-05/003F Environmental Protection Final Agency 8-24-2005 Provisional Peer Reviewed Toxicity Values for 2-Amino-4,6-Dinitrotoluene and 4-Amino-2,6-Dinitrotoluene (CASRNs 35572-78-2 and 19406-51-0) Derivation of Subchronic and Chronic Oral RfDs Superfund Health Risk Technical Support Center National Center for Environmental Assessment Office of Research and Development U.S. Environmental Protection Agency Cincinnati, OH 45268 ------- Acronyms and Abbreviations bw body weight cc cubic centimeters CD Caesarean Delivered CERCLA Comprehensive Environmental Response, Compensation and Liability Act of 1980 CNS central nervous system cu.m cubic meter DWEL Drinking Water Equivalent Level FEL frank-effect level FIFRA Federal Insecticide, Fungicide, and Rodenticide Act g grams GI gastrointestinal HEC human equivalent concentration Hgb hemoglobin i.m. intramuscular i.p. intraperitoneal i.v. intravenous IRIS Integrated Risk Information System IUR inhalation unit risk kg kilogram L liter LEL lowest-effect level LOAEL lowest-observed-adverse-effect level LOAEL(ADJ) LOAEL adjusted to continuous exposure duration LOAEL(HEC) LOAEL adjusted for dosimetric differences across species to a human m meter MCL maximum contaminant level MCLG maximum contaminant level goal MF modifying factor mg milligram mg/kg milligrams per kilogram mg/L milligrams per liter MRL minimal risk level 1 ------- MTD maximum tolerated dose MTL median threshold limit NAAQS National Ambient Air Quality Standards NOAEL no-observed-adverse-effect level NOAEL(ADJ) NOAEL adjusted to continuous exposure duration NOAEL(HEC) NOAEL adjusted for dosimetric differences across species to a human NOEL no-observed-effect level OSF oral slope factor p-IUR provisional inhalation unit risk p-OSF provisional oral slope factor p-RfC provisional inhalation reference concentration p-RfD provisional oral reference dose PBPK physiologically based pharmacokinetic PPb parts per billion ppm parts per million PPRTV Provisional Peer Reviewed Toxicity Value RBC red blood cell(s) RCRA Resource Conservation and Recovery Act RDDR Regional deposited dose ratio (for the indicated lung region) REL relative exposure level RfC inhalation reference concentration RfD oral reference dose RGDR Regional gas dose ratio (for the indicated lung region) s.c. subcutaneous SCE sister chromatid exchange SDWA Safe Drinking Water Act sq.cm. square centimeters TSCA Toxic Substances Control Act UF uncertainty factor Hg microgram |j,mol micromoles voc volatile organic compound 11 ------- 8-24-2005 PROVISIONAL PEER REVIEWED TOXICITY VALUES FOR 2-AMINO-4,6-DINITROTOLUENE (CASRN 35572-78-2) AND 4-AMINO-2,6-DINITROTOLUENE (CASRN 19406-51-0) Derivation of Subchronic and Chronic Oral RfDs Background On December 5, 2003, the U.S. Environmental Protection Agency's (EPA's) Office of Superfund Remediation and Technology Innovation (OSRTI) revised its hierarchy of human health toxicity values for Superfund risk assessments, establishing the following three tiers as the new hierarchy: 1. EPA's Integrated Risk Information System (IRIS). 2. Provisional Peer-Reviewed Toxicity Values (PPRTV) used in EPA's Superfund Program. 3. Other (peer-reviewed) toxicity values, including: ~ Minimal Risk Levels produced by the Agency for Toxic Substances and Disease Registry (ATSDR), ~ California Environmental Protection Agency (CalEPA) values, and ~ EPA Health Effects Assessment Summary Table (HEAST) values. A PPRTV is defined as a toxicity value derived for use in the Superfund Program when such a value is not available in EPA's Integrated Risk Information System (IRIS). PPRTVs are developed according to a Standard Operating Procedure (SOP) and are derived after a review of the relevant scientific literature using the same methods, sources of data, and Agency guidance for value derivation generally used by the EPA IRIS Program. All provisional toxicity values receive internal review by two EPA scientists and external peer review by three independently selected scientific experts. PPRTVs differ from IRIS values in that PPRTVs do not receive the multi-program consensus review provided for IRIS values. This is because IRIS values are generally intended to be used in all EPA programs, while PPRTVs are developed specifically for the Superfund Program. Because science and available information evolve, PPRTVs are initially derived with a three-year life-cycle. However, EPA Regions or the EPA Headquarters Superfund Program sometimes request that a frequently used PPRTV be reassessed. Once an IRIS value for a specific chemical becomes available for Agency review, the analogous PPRTV for that same chemical is retired. It should also be noted that some PPRTV manuscripts conclude that a PPRTV cannot be derived based on inadequate data. 1 ------- 8-24-2005 Disclaimers Users of this document should first check to see if any IRIS values exist for the chemical of concern before proceeding to use a PPRTV. If no IRIS value is available, staff in the regional Superfund and RCRA program offices are advised to carefully review the information provided in this document to ensure that the PPRTVs used are appropriate for the types of exposures and circumstances at the Superfund site or RCRA facility in question. PPRTVs are periodically updated; therefore, users should ensure that the values contained in the PPRTV are current at the time of use. It is important to remember that a provisional value alone tells very little about the adverse effects of a chemical or the quality of evidence on which the value is based. Therefore, users are strongly encouraged to read the entire PPRTV manuscript and understand the strengths and limitations of the derived provisional values. PPRTVs are developed by the EPA Office of Research and Development's National Center for Environmental Assessment, Superfund Health Risk Technical Support Center for OSRTI. Other EPA programs or external parties who may choose of their own initiative to use these PPRTVs are advised that Superfund resources will not generally be used to respond to challenges of PPRTVs used in a context outside of the Superfund Program. Questions Regarding PPRTVs Questions regarding the contents of the PPRTVs and their appropriate use (e.g., on chemicals not covered, or whether chemicals have pending IRIS toxicity values) may 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), or OSRTI. INTRODUCTION RfDs for isomers (C7H7N304) 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6- dinitrotoluene (4-ADNT) are not listed on IRIS (U.S. EPA, 2005), the Drinking Water Standards and Health Advisories table (U.S. EPA, 2002), or the HEAST (U.S. EPA, 1997). No documents for these chemicals are listed in the CARA database (U.S. EPA, 1991, 1994). ATSDR (2002) has not published a Toxicological Profile for 2- or 4-ADNT, and these chemicals are not listed in the NTP (2002) Management Status Reports. IARC (2002) and WHO (2002) have not published reviews for 2- or 4-ADNT, nor have these chemicals been reviewed in Patty's Toxicology (2001). Literature searches of the following databases were conducted from 1965 to August 2002 for relevant studies: TOXLINE, MEDLINE, TSCATS, GENETOX, HSDB, CANCERLIT, CCRIS, RTECS, EMIC/EMICBACK, DART/ETICBACK and BIOSIS. 2 ------- 8-24-2005 An updated literature search was conducted in September 2004 by NCEA-Cincinnati using TOXLINE, MEDLINE, Chemical Abstracts, and Biological Abstracts databases. REVIEW OF PERTINENT LITERATURE Human Studies No data regarding the potential subchronic or chronic oral toxicity of 2- or 4-ADNT in humans were located. 2- and 4-ADNT have been identified as major human metabolites of 2,4,6-trinitrotoluene (TNT), appearing in the urine of TNT-exposed workers (Ahlborg et al., 1988; Almog et al., 1983; Channon et al., 1944; Lemberg and Callaghan, 1944; Woollen et al., 1986; Yinon and Hwang, 1986a, 1987). 2- and 4-ADNT-hemoglobin (Hgb) adducts have also been detected in TNT-exposed workers (Fang et al., 1994; Sabbioni et al., 1996). Animal Studies Available data on the toxicity of 2 and 4-ADNT are limited to acute duration lethality studies. For 2-ADNT, oral LD50s of 1394-2240 and 1522-1722 mg/kg have been reported for the rat and mouse, respectively (Ellis et al., 1980). Rat oral LD50s of 959-1360 and mouse oral LD50s of 1342-1495 mg/kg have been reported for 4-ADNT (Ellis et al., 1980). As in humans, 2- and 4-ADNT have been identified as TNT urinary metabolites in rats (Yinon and Hwang, 1985), rabbits (Yinon and Hwang, 1986b), and dogs (Synder, 1946), and 2- and 4-ADNT-Hgb adducts have been detected in rats exposed to TNT (Liu et al., 1992). PROVISIONAL SUBCHRONIC AND CHRONIC RfDs FOR 2-AMINO-4,6- DINITROTOLUENE AND 4-AMINO-2,6-DINITROTOLUENE A lack of toxicological data on either subchronic or chronic oral exposure to 2-amino-4,6- dinitrotoluene or 4-amino-2,6-dinitrotoluene in humans or animals precludes derivation of a provisional reference dose. REFERENCES Ahlborg, G., Jr., P. Einisto and M. Sorsa. 1988. Mutagenic activity and metabolism in the urine of workers exposed to trinitrotoluene (TNT). Br. J. Ind. Med. 45: 353-358. Almog, J., S. Kraus and A. Basch. 1983. Determination of TNT metabolites in urine. Arch. Toxicol. Suppl. 6:351-353. 3 ------- 8-24-2005 ATSDR (Agency for Toxic Substances and Disease Registry). 2002. Toxicological Profile Information Sheet. Online, http://www.atsdr.cdc.gov/toxpro2.html Channon, H.J., G.T. Mills and R.T. Williams. 1944. The metabolism of 2,4,6-trinitrotoluene (TNT). Biochem. J. 38:70-85. Ellis, H.V., C.-B. Hong and C.-C. Lee. 1980. Mammalian toxicity of munitions compounds. Summary of toxicity of nitrotoluenes. Progress Report No. 11. MRI Project No. 3900-B. Kansas City, MO. DAMD17-74-C-4073. Fang J.-L., Y.-Y. Liu and Y.-W. Wang. 1994. Determination of hemoglobin adducts from the trinitrotoluene-exposed workers by using a competitive inhibition enzyme-linked immunosorbent assay. Zhongguo Yaolixue Yu Dulixue Zazhi. 8:136-138. I ARC (International Agency for Research on Cancer). 2002. IARC Agents and Summary Evaluations. Online. http://193.51.164.ll/cgi/iHound/Chem/iH Chem Frames.html Lemberg, R. and J.P. Callaghan. 1944. Metabolism of symmetrical trinitrotoluene. Nature. 154: 768-769. Liu, Y.-Y., A.Y.H. Lu, R.A. Stearns and S.-H.L. Chiu. 1992. In vivo covalent binding of carbon-14 trinitrotoluene to proteins in the rat. Chem-Biol. Interact. 82: 1-19. NTP (National Toxicology Program). 2002. Management Status Report. Online. http://ntp-server.niehs.nih.gov/ Patty's Toxicology, 5th ed. 2001. E. Bingham, B. Cohrssen and C.H. Powel, Ed. John Wiley and Sons, New York. 9: 1-362. Sabbioni, G., J. Wei and Y.-Y. Liu. 1996. Determination of hemoglobin adducts in workers exposed to 2,4,6-trinitrotoluene. J. Chromatogr. B. Biomed. Appl. 682: 43-248. Synder, R.K. 1946. Metabolites of 2,4,6-trinitrotoluene excreted in urine of dogs. J. Ind. Hyg. Toxicol. 28: 59-77. (Cited in Almog et al., 1983; Yinon and Hwang, 1985; Woollen et al., 1986) U.S. EPA. 1991. Chemical Assessments and Related Activities (CARA). Office of Health and Environmental Assessment, Washington, DC. April. U.S. EPA. 1994. Chemical Assessments and Related Activities (CARA). Office of Health and Environmental Assessment, Washington, DC. December. 4 ------- 8-24-2005 U.S. EPA. 1997. Health Effects Assessment Summary Tables. FY-1997 Update. 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. July. EPA/540/R-97/036. NTIS 97-921199. U.S. EPA. 2002. 2002 Edition of the Drinking Water Standards and Health Advisories. Office of Water, Washington, DC. EPA 822-R-02-038. Online. http://www.epa.gov/waterscience/drinking/standards/dwstandards.pdf U.S. EPA. 2005. Integrated Risk Information System (IRIS). Office of Research and Development, National Center for Environmental Assessment, Washington, DC. Online. http://www.epa.gov/iris/ WHO (World Health Organization). 2002. Online Catalogs for the Environmental Criteria Series. Online. http://193.51.164.ll/cgi/iHound/Chem/iH Chem Frames.html Woollen, B.H., M.G. Hall, R. Craig and G.T. Steel. 1986. Trinirotoluene assessment of occupational absorption during manufacture of explosives. Br. J. Ind. Med. 43: 465-473. Yinon, J. and D.-G. Hwang. 1985. Identification of urinary metabolites of 2,4,6-trinitrotoluene in rats by liquid chromatography-mass spectrometry. Toxicol. Lett. 26: 205-209. Yinon J. and D.-G. Hwang. 1986a. Metabolic studies of explosives 5. Detection and analysis of 2,4,6-trinitrotoluene and its metabolites in urine of munition workers by micro liquid chromatography/mass spectrometry. Biomed. Chromatgr. 1:123-125. Yinon J. and D.-G. Hwang. 1986b. Metabolic studies of explosives. IV. Determination of 2,4,6-trinitrotoluene and its metabolites in blood of rabbits by high-performance liquid chromatography-mass spectrometry. J. Chromatgr. 375: 154-158. Yinon, J. and D.-G. Hwang. 1987. Applications of liquid chromatography-mass spectrometry in metabolic studies of explosives. J. Chromatog. 394: 253-257. 5 ------- 5-17-05 Provisional Peer Reviewed Toxicity Values for 2-Amino-4,6-Dinitrotoluene and 4-Amino-2,6-Dinitrotoluene (CASRNs 35572-78-2 and 19406-51-0) Derivation of Subchronic and Chronic Inhalation RfCs Superfund Health Risk Technical Support Center National Center for Environmental Assessment Office of Research and Development U.S. Environmental Protection Agency Cincinnati, OH 45268 ------- Acronyms and Abbreviations bw body weight cc cubic centimeters CD Caesarean Delivered CERCLA Comprehensive Environmental Response, Compensation and Liability Act of 1980 CNS central nervous system cu.m cubic meter DWEL Drinking Water Equivalent Level FEL frank-effect level FIFRA Federal Insecticide, Fungicide, and Rodenticide Act g grams GI gastrointestinal HEC human equivalent concentration Hgb hemoglobin i.m. intramuscular i.p. intraperitoneal i.v. intravenous IRIS Integrated Risk Information System IUR inhalation unit risk kg kilogram L liter LEL lowest-effect level LOAEL lowest-observed-adverse-effect level LOAEL(ADJ) LOAEL adjusted to continuous exposure duration LOAEL(HEC) LOAEL adjusted for dosimetric differences across species to a human m meter MCL maximum contaminant level MCLG maximum contaminant level goal MF modifying factor mg milligram mg/kg milligrams per kilogram mg/L milligrams per liter MRL minimal risk level 1 ------- MTD maximum tolerated dose MTL median threshold limit NAAQS National Ambient Air Quality Standards NOAEL no-observed-adverse-effect level NOAEL(ADJ) NOAEL adjusted to continuous exposure duration NOAEL(HEC) NOAEL adjusted for dosimetric differences across species to a human NOEL no-observed-effect level OSF oral slope factor p-IUR provisional inhalation unit risk p-OSF provisional oral slope factor p-RfC provisional inhalation reference concentration p-RfD provisional oral reference dose PBPK physiologically based pharmacokinetic PPb parts per billion ppm parts per million PPRTV Provisional Peer Reviewed Toxicity Value RBC red blood cell(s) RCRA Resource Conservation and Recovery Act RDDR Regional deposited dose ratio (for the indicated lung region) REL relative exposure level RfC inhalation reference concentration RfD oral reference dose RGDR Regional gas dose ratio (for the indicated lung region) s.c. subcutaneous SCE sister chromatid exchange SDWA Safe Drinking Water Act sq.cm. square centimeters TSCA Toxic Substances Control Act UF uncertainty factor US microgram |j,mol micromoles VOC volatile organic compound 11 ------- 5-17-05 PROVISIONAL PEER REVIEWED TOXICITY VALUES FOR 2-AMIN0-4,6-DINITR0T0LUENE (CASRN 35572-78-2) AND 4-AMINO-2,6-DINITROTOLUENE (CASRN 19406-51-0) Derivation of Subchronic or Chronic Inhalation RfCs Background On December 5, 2003, the U.S. Environmental Protection Agency's (EPA's) Office of Superfund Remediation and Technology Innovation (OSRTI) revised its hierarchy of human health toxicity values for Superfund risk assessments, establishing the following three tiers as the new hierarchy: 1. EPA's Integrated Risk Information System (IRIS). 2. Provisional Peer-Reviewed Toxicity Values (PPRTV) used in EPA's Superfund Program. 3. Other (peer-reviewed) toxicity values, including: ~ Minimal Risk Levels produced by the Agency for Toxic Substances and Disease Registry (ATSDR), ~ California Environmental Protection Agency (CalEPA) values, and ~ EPA Health Effects Assessment Summary Table (HEAST) values. A PPRTV is defined as a toxicity value derived for use in the Superfund Program when such a value is not available in EPA's Integrated Risk Information System (IRIS). PPRTVs are developed according to a Standard Operating Procedure (SOP) and are derived after a review of the relevant scientific literature using the same methods, sources of data, and Agency guidance for value derivation generally used by the EPA IRIS Program. All provisional toxicity values receive internal review by two EPA scientists and external peer review by three independently selected scientific experts. PPRTVs differ from IRIS values in that PPRTVs do not receive the multi-program consensus review provided for IRIS values. This is because IRIS values are generally intended to be used in all EPA programs, while PPRTVs are developed specifically for the Superfund Program. Because science and available information evolve, PPRTVs are initially derived with a three-year life-cycle. However, EPA Regions (or the EPA HQ Superfund Program) sometimes request that a frequently used PPRTV be reassessed. Once an IRIS value for a specific chemical becomes available for Agency review, the analogous PPRTV for that same chemical is retired. It should also be noted that some PPRTV manuscripts conclude that a PPRTV cannot be derived based on inadequate data. 1 ------- 5-17-05 Disclaimers Users of this document should first check to see if any IRIS values exist for the chemical of concern before proceeding to use a PPRTV. If no IRIS value is available, staff in the regional Superfund and RCRA program offices are advised to carefully review the information provided in this document to ensure that the PPRTVs used are appropriate for the types of exposures and circumstances at the Superfund site or RCRA facility in question. PPRTVs are periodically updated; therefore, users should ensure that the values contained in the PPRTV are current at the time of use. It is important to remember that a provisional value alone tells very little about the adverse effects of a chemical or the quality of evidence on which the value is based. Therefore, users are strongly encouraged to read the entire PPRTV manuscript and understand the strengths and limitations of the derived provisional values. PPRTVs are developed by the EPA Office of Research and Development's National Center for Environmental Assessment, Superfund Health Risk Technical Support Center for OSRTI. Other EPA programs or external parties who may choose of their own initiative to use these PPRTVs are advised that Superfund resources will not generally be used to respond to challenges of PPRTVs used in a context outside of the Superfund Program. Questions Regarding PPRTVs Questions regarding the contents of the PPRTVs and their appropriate use (e.g., on chemicals not covered, or whether chemicals have pending IRIS toxicity values) may 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), or OSRTI. INTRODUCTION RfCs for isomers (C7H7N304) 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6- dinitrotoluene (4-ADNT) are not listed on IRIS (U.S. EPA, 2002) or the HEAST (U.S. EPA, 1997). No documents for these chemicals are listed in the CARA database (U.S. EPA, 1991, 1994). ATSDR (2002) has not published a Toxicological Profile for 2- or 4-ADNT and these chemicals were not listed in the NTP (2002) Management Status Reports. ACGIH (2002), NIOSH (2002) and OSHA (2002a,b) have not established occupational exposure limits for these chemicals. IARC (2002) and WHO (2002) have not published reviews for 2- or 4-ADNT, nor have these chemicals been reviewed in Patty's Toxicology (2001). Literature searches of the following databases were conducted from 1965 to August 2002 for relevant studies: TOXLINE, MEDLINE, TSCATS, GENETOX, HSDB, CANCERLIT, CCRIS, RTECS, EMIC/EMICBACK, DART/ETICBACK and BIOSIS. Additional literature searches from August 2002 through 2 ------- 5-17-05 October 2004 were conducted by NCEA-Cincinnati using MEDLINE, TOXLINE, Chemical Abstracts, and Biological Abstracts databases. REVIEW OF PERTINENT LITERATURE Human Studies No data regarding the potential subchronic or chronic inhalation toxicity of 2- or 4-ADNT in humans were located. 2- and 4-ADNT have been identified as major human metabolites of 2,4,6-trinitrotoluene (TNT), appearing in the urine of TNT-exposed workers (Ahlborg et al., 1988; Almog et al., 1983; Channon et al., 1944; Lemberg and Callaghan, 1944; Woollen et al., 1986; Yinon and Hwang, 1986a, 1987). 2- and 4-ADNT-hemoglobin (Hb) adducts have also been detected in TNT-exposed workers (Fang et al., 1994; Sabbioni et al., 1996). Animal Studies No data regarding the toxicity of 2- or 4-ADNT in animals following subchronic or chronic inhalation exposure were located. As in humans, 2- and 4-ADNT have been identified as TNT urinary metabolites in rats (Yinon and Hwang, 1985), rabbits (Yinon and Hwang, 1986b), and dogs (Synder, 1946), and 2- and 4-ADNT-Hb adducts have been detected in rats exposed to TNT (Liu et al., 1992). DERIVATION OF A PROVISIONAL SUBCHRONIC OR CHRONIC RfC FOR 2-AMIN0-4,6-DINITR0T0LUENE AND 4-AMINO-2,6-DINITROTOLUENE The toxicity data for the ADNTs are insufficient for derivation of p-RfCs. REFERENCES ACGIH (American Conference of Governmental Industrial Hygienists). 2002. TLVsŪ and BEIsŪ: Threshold Limit Values for Chemical Substances and Physical Agents, Biological Exposure Indices. Cincinnati, OH. Ahlborg, G., Jr., P. Einisto and M. Sorsa. 1988. Mutagenic activity and metabolism in the urine of workers exposed to trinitrotoluene (TNT). Br. J. Ind. Med. 45: 353-358. Almog, J., S. Kraus and A. Basch. 1983. Determination of TNT metabolites in urine. Arch. Toxicol. Suppl. 6: 351-353. 3 ------- 5-17-05 ATSDR (Agency for Toxic Substances and Disease Registry). 2002. Toxicological Profile Information Sheet. Online, http://www.atsdr.cdc.gov/toxpro2.html Channon, H.J., G.T. Mills and R.T. Williams. 1944. The metabolism of 2,4,6-trinitrotoluene (TNT). Biochem. J. 38: 70-85. Fang J.-L., Y.-Y. Liu and Y.-W. Wang. 1994. Determination of hemoglobin adducts from the trinitrotoluene-exposed workers by using a competitive inhibition enzyme-linked immunosorbent assay. Zhongguo Yaolixue Yu Dulixue Zazhi. 8:136-138. IARC (International Agency for Research on Cancer). 2002. IARC Agents and Summary Evaluations. Online. http://193.51.164.il/cgi/iHound/Chem/iH Chem Frames.html Lemberg, R. and J.P. Callaghan. 1944. Metabolism of symmetrical trinitrotoluene. Nature. 154: 768-769. Liu, Y.-Y., A.Y.H. Lu, R.A. Stearns and S.-H.L. Chiu. 1992. In vivo covalent binding of carbon-14 trinitrotoluene to proteins in the rat. Chem-Biol. Interact. 82: 1-19. NIOSH (National Institute for Occupational Safety and Health). 2002. NIOSH Pocket Guide to Chemical Hazards. Online. http://www.cdc.gOv/niosh/npg/.html NTP (National Toxicology Program). 2002. Management Status Report. Online. http://ntp-server.niehs.nih.gov/ OSHA (Occupational Safety and Health Administration). 2002a. OSHA Standard 1910.1000 Table Z-l. Part Z, Toxic and Hazardous Substances. Online. http://www.osha-slc.gov/OshStd data/1910 1000 TABLE Z-l.html OSHA (Occupational Safety and Health Administration). 2002b. OSHA Standard 1915.1000 for Air Contaminants. Part Z, Toxic and Hazardous Substances. Online. http://www.osha-slc.gov/OshStd data/1915 1000.html Patty's Toxicology, 5th ed. 2001. E. Bingham, B. Cohrssen and C.H. Powel, Ed. John Wiley and Sons, New York. 9: 1-362. Sabbioni, G., J. Wei and Y-Y. Liu. 1996. Determination of hemoglobin adducts in workers exposed to 2,4,6-trinitrotoluene. J. Chromatogr. B. Biomed. Appl. 682: 43-248. 4 ------- 5-17-05 Synder, R.K. 1946. Metabolites of 2,4,6-trinitrotoluene excreted in urine of dogs. J. Ind. Hyg. Toxicol. 28: 59-77. (Cited in Almong et al., 1983; Yinon and Hwang, 1985; Woollen et al., 1986) U.S. EPA. 1991. Chemical Assessments and Related Activities (CARA). Office of Health and Environmental Assessment, Washington, DC. April. U.S. EPA. 1994. Chemical Assessments and Related Activities (CARA). Office of Health and Environmental Assessment, Washington, DC. December. U.S. EPA. 1997. Health Effects Assessment Summary Tables. FY-1997 Update. 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. July. EPA/540/R-97/036. NTIS 97-921199. U.S. EPA. 2002. Integrated Risk Information System (IRIS). Office of Research and Development, National Center for Environmental Assessment, Washington, DC. Online. http://www.epa.gov/iris/ WHO (World Health Organization). 2002. Online Catalogs for the Environmental Criteria Series. Online. http://193.51.164.il/cgi/iHound/Chem/iH Chem Frames.html Woollen, B.H., M.G. Hall, R. Craig and G.T. Steel. 1986. Trinirotoluene assessment of occupational absorption during manufacture of explosives. Br. J. Ind. Med. 43: 465-473. Yinon, J. and D.-G. Hwang. 1985. Identification of urinary metabolites of 2,4,6-trinitrotoluene in rats by liquid chromatography-mass spectrometry. Toxicol. Lett. 26: 205-209. Yinon J. and D.-G. Hwang. 1986a. Metabolic studies of explosives 5. Detection and analysis of 2,4,6-trinitrotoluene and its metabolites in urine of munition workers by micro liquid chromatography/mass spectrometry. Biomed. Chromatgr. 1:123-125. Yinon J. and D.-G. Hwang. 1986b. Metabolic studies of explosives. IV. Determination of 2,4,6-trinitrotoluene and its metabolites in blood of rabbits by high-performance liquid chromatography-mass spectrometry. J. Chromatgr. 375: 154-158. Yinon, J. and D.-G. Hwang. 1987. Applications of liquid chromatography-mass spectrometry in metabolic studies of explosives. J. Chromatog. 394: 253-257. 5 ------- 5-17-05 Provisional Peer Reviewed Toxicity Values for 2-Amino-4,6-Dinitrotoluene and 4-Amino-2,6-Dinitrotoluene (CASRNs 35572-78-2 and 19406-51-0) Derivation of a Carcinogenicity Assessment Superfund Health Risk Technical Support Center National Center for Environmental Assessment Office of Research and Development U.S. Environmental Protection Agency Cincinnati, OH 45268 ------- Acronyms and Abbreviations bw body weight cc cubic centimeters CD Caesarean Delivered CERCLA Comprehensive Environmental Response, Compensation and Liability Act of 1980 CNS central nervous system cu.m cubic meter DWEL Drinking Water Equivalent Level FEL frank-effect level FIFRA Federal Insecticide, Fungicide, and Rodenticide Act g grams GI gastrointestinal HEC human equivalent concentration Hgb hemoglobin i.m. intramuscular i.p. intraperitoneal i.v. intravenous IRIS Integrated Risk Information System IUR inhalation unit risk kg kilogram L liter LEL lowest-effect level LOAEL lowest-observed-adverse-effect level LOAEL(ADJ) LOAEL adjusted to continuous exposure duration LOAEL(HEC) LOAEL adjusted for dosimetric differences across species to a human m meter MCL maximum contaminant level MCLG maximum contaminant level goal MF modifying factor mg milligram mg/kg milligrams per kilogram mg/L milligrams per liter MRL minimal risk level 1 ------- MTD maximum tolerated dose MTL median threshold limit NAAQS National Ambient Air Quality Standards NOAEL no-observed-adverse-effect level NOAEL(ADJ) NOAEL adjusted to continuous exposure duration NOAEL(HEC) NOAEL adjusted for dosimetric differences across species to a human NOEL no-observed-effect level OSF oral slope factor p-IUR provisional inhalation unit risk p-OSF provisional oral slope factor p-RfC provisional inhalation reference concentration p-RfD provisional oral reference dose PBPK physiologically based pharmacokinetic PPb parts per billion ppm parts per million PPRTV Provisional Peer Reviewed Toxicity Value RBC red blood cell(s) RCRA Resource Conservation and Recovery Act RDDR Regional deposited dose ratio (for the indicated lung region) REL relative exposure level RfC inhalation reference concentration RfD oral reference dose RGDR Regional gas dose ratio (for the indicated lung region) s.c. subcutaneous SCE sister chromatid exchange SDWA Safe Drinking Water Act sq.cm. square centimeters TSCA Toxic Substances Control Act UF uncertainty factor Hg microgram |j,mol micromoles VOC volatile organic compound 11 ------- 5-17-05 PROVISIONAL PEER REVIEWED TOXICITY VALUES FOR 2-AMIN0-4,6-DINITR0T0LUENE (CASRN 35572-78-2) AND 4-AMINO-2,6-DINITROTOLUENE (CASRN 19406-51-0) Derivation of a Carcinogenicity Assessment Background On December 5, 2003, the U.S. Environmental Protection Agency's (EPA's) Office of Superfund Remediation and Technology Innovation (OSRTI) revised its hierarchy of human health toxicity values for Superfund risk assessments, establishing the following three tiers as the new hierarchy: 1. EPA's Integrated Risk Information System (IRIS). 2. Provisional Peer-Reviewed Toxicity Values (PPRTV) used in EPA's Superfund Program. 3. Other (peer-reviewed) toxicity values, including: ~ Minimal Risk Levels produced by the Agency for Toxic Substances and Disease Registry (ATSDR), ~ California Environmental Protection Agency (CalEPA) values, and ~ EPA Health Effects Assessment Summary Table (HEAST) values. A PPRTV is defined as a toxicity value derived for use in the Superfund Program when such a value is not available in EPA's Integrated Risk Information System (IRIS). PPRTVs are developed according to a Standard Operating Procedure (SOP) and are derived after a review of the relevant scientific literature using the same methods, sources of data, and Agency guidance for value derivation generally used by the EPA IRIS Program. All provisional toxicity values receive internal review by two EPA scientists and external peer review by three independently selected scientific experts. PPRTVs differ from IRIS values in that PPRTVs do not receive the multi-program consensus review provided for IRIS values. This is because IRIS values are generally intended to be used in all EPA programs, while PPRTVs are developed specifically for the Superfund Program. Because science and available information evolve, PPRTVs are initially derived with a three-year life-cycle. However, EPA Regions (or the EPA HQ Superfund Program) sometimes request that a frequently used PPRTV be reassessed. Once an IRIS value for a specific chemical becomes available for Agency review, the analogous PPRTV for that same chemical is retired. It should also be noted that some PPRTV manuscripts conclude that a PPRTV cannot be derived based on inadequate data. 1 ------- 5-17-05 Disclaimers Users of this document should first check to see if any IRIS values exist for the chemical of concern before proceeding to use a PPRTV. If no IRIS value is available, staff in the regional Superfund and RCRA program offices are advised to carefully review the information provided in this document to ensure that the PPRTVs used are appropriate for the types of exposures and circumstances at the Superfund site or RCRA facility in question. PPRTVs are periodically updated; therefore, users should ensure that the values contained in the PPRTV are current at the time of use. It is important to remember that a provisional value alone tells very little about the adverse effects of a chemical or the quality of evidence on which the value is based. Therefore, users are strongly encouraged to read the entire PPRTV manuscript and understand the strengths and limitations of the derived provisional values. PPRTVs are developed by the EPA Office of Research and Development's National Center for Environmental Assessment, Superfund Health Risk Technical Support Center for OSRTI. Other EPA programs or external parties who may choose of their own initiative to use these PPRTVs are advised that Superfund resources will not generally be used to respond to challenges of PPRTVs used in a context outside of the Superfund Program. Questions Regarding PPRTVs Questions regarding the contents of the PPRTVs and their appropriate use (e.g., on chemicals not covered, or whether chemicals have pending IRIS toxicity values) may 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), or OSRTI. INTRODUCTION Carcinogenicity assessments for isomers (C7H7N304) 2-amino-4,6-dinitrotoluene (2- ADNT) and 4-amino-2,6-dinitrotoluene (4-ADNT) are not listed on IRIS (U.S. EPA, 2002a), the Drinking Water Standards and Health Advisories table (U.S. EPA, 2002b), or the HEAST (U.S. EPA, 1997). No documents for these chemicals are listed in the CARA database (U.S. EPA, 1991, 1994). ATSDR (2002) has not published a Toxicological Profile for 2- or 4-ADNT and these chemicals were not listed in the NTP (2002) Management Status Reports. ACGIH (2002), IARC (2002) and WHO (2002) have not assessed the carcinogenicity of 2- or 4-ADNT, nor have these chemicals been reviewed in Patty's Toxicology (2001). Literature searches of the following databases were conducted from 1965 to August 2002 for relevant studies: TOXLINE, MEDLINE, TSCATS, GENETOX, HSDB, CANCERLIT, CCRIS, RTECS, EMIC/EMICBACK, DART/ETICBACK and BIOSIS. Additional literature searches from August 2002 through 2 ------- 5-17-05 October 2004 were conducted by NCEA-Cincinnati using MEDLINE, TOXLINE, Chemical Abstracts, and Biological Abstracts databases. REVIEW OF PERTINENT LITERATURE Human Studies No data regarding the potential carcinogenicity of 2- or 4-ADNT in humans were located. 2- and 4-ADNT have been identified as major human metabolites of 2,4,6-trinitrotoluene (TNT), appearing in the urine of TNT-exposed workers (Ahlborg et al., 1988; Almog et al., 1983; Channon et al., 1944; Lemberg and Callaghan, 1944; Woollen et al., 1986; Yinon and Hwang, 1986a, 1987). 2- and 4-ADNT-hemoglobin (Hb) adducts have also been detected in TNT- exposed workers (Fang et al., 1994; Sabbioni et al., 1996). Animal Studies No data regarding the carcinogenicity of 2- or 4-ADNT in animals following subchronic or chronic inhalation exposure were located. As in humans, 2- and 4-ADNT have been identified as TNT urinary metabolites in rats (Yinon and Hwang, 1985), rabbits (Yinon and Hwang, 1986b), and dogs (Synder, 1946), and 2- and 4-ADNT-Hb adducts have been detected in rats exposed to TNT (Liu et al., 1992). Other Studies Ahlborg et al. (1988) tested the urine of 41 TNT-exposed workers in a munitions plant for mutagenicity using Salmonella typhimurium strains TA98 and TA98 NR (deficient in endogenous nitroreductase activity) without exogenous metabolic activation. Mutagenic activity in both strains was significantly correlated with urine 4-ADNT concentration, but not with TNT or 2-ADNT concentration. The authors contended that nitroreductase was not significantly contributory to the mutagenicity. In another study, urine from TNT-treated rats was separated into fractions containing predominantly TNT or ADNT and tested for mutagenicity in the Ames S. typhimurium mutagenicity assay without metabolic activation (Brooks et al., 1997). The TNT fraction was positive in four tester strains: TA98, TA98NR, TA100, and TA100NR. The ADNT fraction (containing 2- and 4-ADNT) tested positive in TA98, but negative in the remaining three strains. Spanggord et al. (1982, 1995) tested the mutagenicity of numerous polynitroaromatic chemicals in six strains of S. typhimurium, including a nitroreductase deficient strain, both with and without metabolic activation. 4-ADNT had lower "mutagenic potency" than 2-ADNT, which was lower than TNT. Neither 2-ADNT, 4-ADNT nor TNT were mutagenic in the nitroreductase-deficient strain. A similar pattern was seen in most other studies, with positive results reported in at least one strain and potency increasing from 4-ADNT to 2-ADNT to TNT 3 ------- 5-17-05 (Tan et al., 1992; Honeycutt et al., 1996; Lachance et al., 1999; Karamova et al., 1994). One study reported negative results with 2-ADNT and 4-ADNT with or without activation, and positive results with TNT only in TA98 without activation (Won et al., 1976). Banerjee and Dutta (1997) found that exposure of NG108 neuroblastoma and MCF7 breast cancer cells to 2-ADNT causes p53 gene mutation and induces apoptotic changes. PROVISIONAL WEIGHT-OF-EVIDENCE CLASSIFICATION No studies of the potential carcinogenicity of 2- or 4-ADNT in humans or animals were located. Genotoxicity data indicate that both 2- and 4-ADNT have some mutagenic activity (Salmonella typhimurium strains TA98 only) but not in the nitroreductase deficient strain. According to the Guidelines for Carcinogen Risk Assessment (U.S. EPA, 2005), the data for these chemicals are inadequate for an assessment of human carcinogenic potential. QUANTITATIVE ESTIMATES OF CARCINOGENIC RISK Lack of oral carcinogenic data and pharmacokinetic/pharmacodynamic information precludes derivation of an oral slope factor for both 2-amino-4,6-dinitrotoluene and 4-amino-2,6- dinitro toluene. REFERENCES ACGIH (American Conference of Governmental Industrial Hygienists). 2002. TLVsŪ and BEIsŪ: Threshold Limit Values for Chemical Substances and Physical Agents, Biological Exposure Indices. Cincinnati, OH. Ahlborg, G., Jr., P. Einisto and M. Sorsa. 1988. Mutagenic activity and metabolism in the urine of workers exposed to trinitrotoluene (TNT). Br. J. Ind. Med. 45: 353-358. Almog, J., S. Kraus and A. Basch. 1983. Determination of TNT metabolites in urine. Arch. Toxicol. Suppl. 6: 351-353. ATSDR (Agency for Toxic Substances and Disease Registry). 2002. Toxicological Profile Information Sheet. Online, http://www.atsdr.cdc.gov/toxpro2.html Banerjee, H. and S.K. Dutta. 1997. 2-Amino-4 6-dinitrotoluene exposure to mammalian cells causes p53 gene mutation and induces apoptotic changes. 17th Int. Congr. Biochem. Mol. Biol. 4 ------- 5-17-05 Ann. Mtg. Amer. Soc. Biochem. Mol. Biol. San Francisco, CA, August 24-29, 1997. FASEB. J. 11: A1239. Brooks, L.R., R.W. Jacobson, S.H. Warren et al. 1997. Mutagenicity of HPLC-fractionated urinary metabolites from 2,4,6-trinitrotoluene-treated Fischer 344 rats. Environ. Mol. Mutagen. 30: 298-302. Channon, H.J., G.T. Mills and R.T. Williams. 1944. The metabolism of 2,4,6-trinitrotoluene (TNT). Biochem. J. 38: 70-85. Fang J.-L., Y.-Y. Liu and Y.-W. Wang. 1994. Determination of hemoglobin adducts from the trinitrotoluene-exposed workers by using a competitive inhibition enzyme-linked immunosorbent assay. Zhongguo Yaolixue Yu Dulixue Zazhi. 8:136-138. Honeycutt, M.E., A.S. Jarvis and V.A. McFarland. 1996. Cytotoxicity and mutagenicity of 2,4,6-trinitrotoluene and its metabolites. Ecotoxicol. Environ. Saf. 35: 282-287. IARC (International Agency for Research on Cancer). 2002. IARC Agents and Summary Evaluations. Online. http://193.51.164.il/cgi/iHound/Chem/iH Chem Frames.html Karamova, N.S., O.N. Il'inskaya and O.B. Ivanchenko. 1994. Mutagenic activity of 2,4,6- trinitrotoluene: the role of metabolizing enzymes. Rus. J. Genet. 30: 785-788. (Cited in Lachance et al., 1999) Lachance, B., P. Robidoux and J. Hawari. 1999. Cytotoxic and genotoxic effects of energetic compounds on bacterial and mammalian cells in vitro. Mutat. Res. 444: 25-39. Lemberg, R. and J.P. Callaghan. 1944. Metabolism of symmetrical trinitrotoluene. Nature. 154: 768-769. Liu, Y.-Y., A.Y.H. Lu, R.A. Stearns and S.-H.L. Chiu. 1992. In vivo covalent binding of carbon-14 trinitrotoluene to proteins in the rat. Chem-Biol. Interact. 82: 1-19. NTP (National Toxicology Program). 2002. Management Status Report. Online. http://ntp-server.niehs.nih.gov/ Patty's Toxicology, 5th ed. 2001. E. Bingham, B. Cohrssen and C.H. Powel, Ed. John Wiley and Sons, New York. 9: 1-362. Sabbioni, G., J. Wei and Y.-Y. Liu. 1996. Determination of hemoglobin adducts in workers exposed to 2,4,6-trinitrotoluene. J. Chromatogr. B. Biomed. Appl. 682: 43-248. 5 ------- 5-17-05 Spanggord, R.J., K.E. Mortelmans, A.F. Griffin and V.F. Simmon. 1982. Mutagenicity in Salmonella typhimurium and structure-activity relationship of wastewater components emanating from the manufacture of trinitrotoluene. Environ. Mutagen. 4:163-179. Spanggord, R.J., K.R. Stewart and E.S. Riccio. 1995. Mutagenicity of tetranitroazoxytoluenes: a preliminary screening in Salmonella typhimurium strains TA100 and TA100NR. Mutat. Res. 335:207-211. Synder, R.K. 1946. Metabolites of 2,4,6-trinitrotoluene excreted in urine of dogs. J. Ind. Hyg. Toxicol. 28: 59-77 (Cited in Almong et al., 1983; Yinon and Hwang, 1985; Woollen et al., 1986) Tan, E.L., C.H. Ho, W.H. Griest and R.L. Tyndall. 1992. Mutagenicity of trinitrotoluene and its metabolites formed during composting. J. Toxicol. Environ. Health. 36: 165-175. U.S. EPA. 1991. Chemical Assessments and Related Activities (CARA). Office of Health and Environmental Assessment, Washington, DC. April. U.S. EPA. 1994. Chemical Assessments and Related Activities (CARA). Office of Health and Environmental Assessment, Washington, DC. December. U.S. EPA. 1997. Health Effects Assessment Summary Tables. FY-1997 Update. 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. July. EPA/540/R-97/036. NTIS 97-921199. U.S. EPA. 2002a. Integrated Risk Information System (IRIS). Office of Research and Development, National Center for Environmental Assessment, Washington, DC. Online. http://www.epa.gov/iris/ U.S. EPA. 2002b. 2002 Edition of the Drinking Water Standards and Health Advisories. Office of Water, Washington, DC. EPA 822-R-02-038. Online. http://www.epa. gov/waterscience/drinking/standards/dwstandards .pdf U.S. EPA. 2005. Guidelines for Carcinogen Risk Assessment. Office of Research and Development, National Center for Environmental Assessment, Washington, DC. EPA/63 0/P03/00 IF. WHO (World Health Organization). 2002. Online Catalogs for the Environmental Criteria Series. Online. http://193.51.164.il/cgi/iHound/Chem/iH Chem Frames.html 6 ------- 5-17-05 Won, W.D., L.H. DiSalvo and J. Ng. 1976. Toxicity and mutagenicity of 2,4,6-trinitrotoluene and its microbial metabolites. Appl. Environ. Microbiol. 31: 576-580. Woollen, B.H., M.G. Hall, R. Craig and G.T. Steel. 1986. Trinitrotoluene assessment of occupational absorption during manufacture of explosives. Br. J. Ind. Med. 43: 465-473. Yinon, J. and D.-G. Hwang. 1985. Identification of urinary metabolites of 2,4,6-trinitrotoluene in rats by liquid chromatography-mass spectrometry. Toxicol. Lett. 26: 205-209. Yinon J. and D.-G. Hwang. 1986a. Metabolic studies of explosives 5. Detection and analysis of 2,4,6-trinitrotoluene and its metabolites in urine of munition workers by micro liquid chromatography/mass spectrometry. Biomed. Chromatgr. 1:123-125. Yinon J. and D.-G. Hwang. 1986b. Metabolic studies of explosives. IV. Determination of 2,4,6-trinitrotoluene and its metabolites in blood of rabbits by high-performance liquid chromatography-mass spectrometry. J. Chromatgr. 375: 154-158. Yinon, J. and D.-G. Hwang. 1987. Applications of liquid chromatography-mass spectrometry in metabolic studies of explosives. J. Chromatog. 394: 253-257. 7 ------- |