¦IPs United States Environmental Protection Jr mAgency EPA/690/R-04/001F Final 12-20-2004 Provisional Peer Reviewed Toxicity Values for Alizarin Red Compounds (Various CASRNs) 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 ------- 12-20-04 PROVISIONAL PEER REVIEWED TOXICITY VALUES FOR ALIZARIN RED COMPOUNDS (VARIOUS CASRNS) 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 ------- 12-20-04 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 In the chemical literature, several compounds share the common chemical names "alizarin," "alizarin red" or "alizarine red." Seven distinct chemicals were identified in searches of the CAS Registry. Four of the alizarin chemicals are anthraquinone derivatives, two are azo dyes, and one is of an unknown molecular and structural formula. The alizarin red compounds identified include: Anthraquinone derivatives: 1. alizarin, alizarin red, l,2-dihydroxy-9,10-anthracenedione, or l,2-dihydroxy-9,10-anthraquinone (CASRN 72-48-0); 2 ------- 12-20-04 2. alizarin red S, acid red alizarin, [9,10-dihydro-3,4-dihydroxy-9,10-dioxo-2- anthracenesulfonic acid, monosodium salt], or sodium 3,4-dihydroxyanthraquinone-2- sulfonate (CASRN 130-22-3); 3. alizarine red 3 WS or 9,10-dihydro-3,4,7-trihydro-9,10-dioxo-2-anthracenesulfonic acid, monosodium salt (CASRN 6486-94-8); 4. alizarine red YCAP, l,2,6-trihydroxy-9,10-anthracenedione, or 1,2,6-trihydroxyanthraquinone (CASRN 82-29-1). Azo dyes: 5. acid alizarin red B or 2-[(2-hydroxy-3,6-disulfo-l-naphthalenyl)azo]-benzoic acid, trisodium salt (CASRN 1836-22-2); 6. alizarine chrome red G, acid red 97, or 4,4'-bis[(2-hydroxy-l-naphthalenyl)azo]-[l,r- biphenyl]-2,2'-disulfonic acid, disodium salt (CASRN 10169-02-5). Unknown structural and molecular formula: 7. alizarine red V2A or V2A alizarine red (molecular and structural formula unspecified) (CASRN 12794-90-0). Subchronic or chronic RfDs for the alizarin red compounds identified above are not available on IRIS (U.S. EPA, 2002a), the HEAST (U.S. EPA, 1997) or the Drinking Water Standards and Health Advisories list (U.S. EPA, 2002b). The CARA list (U.S. EPA, 1991, 1994) includes no documents for these chemicals, and these chemicals have not been reviewed by ATSDR (2002), IARC (2002) or WHO (2002). The NTP (2002) Management Status Report provided no relevant information. In June 1996, literature searches were conducted from 1965 to May 1996 in TOXLINE, CANCERLIT, MEDLINE, GENETOX, HSDB, EMIC/EMICBACK, DART/ETICBACK, RTECS and TSCATS for relevant studies. In September 2002, update literature searches were conducted in these databases going back to the previous search in 1996. Additional literature search was conducted by NCEA-Cin from October 2003 through May 2004 using TOXLINE, MEDLINE, Chemical and Biological Abstract databases. 3 ------- 12-20-04 REVIEW OF PERTINENT LITERATURE Human Studies No reports were located regarding the subchronic or chronic toxicity of any of the alizarin red compounds in humans by oral exposure. Animal Studies No reports were located regarding the subchronic or chronic toxicity of any of the alizarin red compounds in animals by oral exposure. Other Studies The only toxicity data available for the alizarin red compounds are a few acute studies and case reports. For example, data from acute rabbit studies suggest that alizarin red (CASRN 72-48-0) is a skin sensitizer (Greenburg and Lester, 1954) and a mild eye irritant (500 mg/24 hours) (NIOSH, 2002). In humans, two case reports of contact dermatitis from the perennial herbs Rubia tinctorum and R. peregrina, both of which contain alizarin red (CASRN 72-48-0) naturally, were published by Castelain and Ducombs (1988). An acute toxicity study in mice is available for alizarin red S (CASRN 130-22-3) wherein an intravenous LD50 of 70 mg/kg was reported (NIOSH, 2002). No reports regarding acute, subchronic or chronic toxicity in humans or animals by any route were located for any of the other alizarin red compounds. FEASIBILITY OF DERIVING A PROVISIONAL SUBCHRONIC OR CHRONIC RfD FOR ALIZARIN RED COMPOUNDS No pertinent data regarding the oral toxicity of alizarin red compounds in humans or experimental animals were located, precluding derivation of subchronic or chronic p-RfDs for alizarin red compounds. REFERENCES ATSDR (Agency for Toxic Substances and Disease Registry). 2002. Toxicological Profile Information Sheet. Online, http://www.atsdr.cdc.gov/toxpro2.html Castelain, M. and G. Ducombs. 1988. Contact dermatitis from madder. Con. Derm. 19: 228-229. 4 ------- 12-20-04 Greenburg, L.A. and D. Lester. 1954. Handbook of Cosmetic Materials. Interscience, New York. (Cited in Castelain and Ducombs, 1988) IARC (International Agency for Research on Cancer). 2002. Monographs Database. Online. http://www-cie.iarc.fr/ NIOSH (National Institute for Occupational Safety and Health). 2002. RTECS (Registry of Toxic Effects of Chemical Substances). Cincinnati, OH. NTP (National Toxicology Program). 2002. Management Status Report. Online. http://ntp-server.niehs.nih.gov/ 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 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 PB97-921199. 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 WHO (World Health Organization). 2002. Online catalogs for the Environmental Health Criteria series. Online, http://www.inchem.org/ehc.html 5 ------- 12-20-04 Provisional Peer Reviewed Toxicity Values for Alizarin Red Compounds (Various CASRNs) 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 Hg microgram |j,mol micromoles voc volatile organic compound 11 ------- 12-20-04 PROVISIONAL PEER REVIEWED TOXICITY VALUES FOR ALIZARIN RED COMPOUNDS (VARIOUS CASRNS) Derivation of Subchronic and 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 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 ------- 12-20-04 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 In the chemical literature, several compounds share the common chemical names "alizarin," "alizarin red" or "alizarine red." Seven distinct chemicals were identified in searches of the CAS Registry. Four of the alizarin chemicals are anthraquinone derivatives, two are azo dyes, and one is of an unknown molecular and structural formula. The alizarin red compounds identified include: Anthraquinone derivatives: 1. alizarin, alizarin red, l,2-dihydroxy-9,10-anthracenedione, or l,2-dihydroxy-9,10-anthraquinone (CASRN 72-48-0); 2 ------- 12-20-04 2. alizarin red S, acid red alizarin, [9,10-dihydro-3,4-dihydroxy-9,10-dioxo-2- anthracenesulfonic acid, monosodium salt], or sodium 3,4-dihydroxyanthraquinone-2- sulfonate (CASRN 130-22-3); 3. alizarine red 3 WS or 9,10-dihydro-3,4,7-trihydro-9,10-dioxo-2-anthracenesulfonic acid, monosodium salt (CASRN 6486-94-8); 4. alizarine red YCAP, l,2,6-trihydroxy-9,10-anthracenedione, or 1,2,6-trihydroxyanthraquinone (CASRN 82-29-1). Azo dyes: 5. acid alizarin red B or 2-[(2-hydroxy-3,6-disulfo-l-naphthalenyl)azo]-benzoic acid, trisodium salt (CASRN 1836-22-2); 6. alizarine chrome red G, acid red 97, or 4,4'-bis[(2-hydroxy-l-naphthalenyl)azo]-[l,r- biphenyl]-2,2'-disulfonic acid, disodium salt (CASRN 10169-02-5). Unknown structural and molecular formula: 7. alizarine red V2A or V2A alizarine red (molecular and structural formula unspecified) (CASRN 12794-90-0). Subchronic or chronic RfCs for the alizarin red compounds identified above are not available on IRIS (U.S. EPA, 2002) or in the HEAST (U.S. EPA, 1997). The CARA list (U.S. EPA, 1991, 1994) includes no documents for these chemicals. These compounds have not been reviewed by ATSDR (2002), IARC (2002) or WHO (2002). Occupational exposure limits have not been derived by ACGIH (2001), NIOSH (2002a) or OSHA (2002). The NTP (2002) Management Status Report provided no relevant information. In June 1996, literature searches were conducted from 1965 to May 1996 in TOXLINE, CANCERLIT, MEDLINE, GENETOX, HSDB, EMIC/EMICBACK, DART/ETICBACK, RTECS and TSCATS for relevant studies. In September 2002, update literature searches were conducted in these databases going back to the previous search in 1996. Additional literature search was conducted by NCEA-Cin from October 2003 through May 2004 using TOXLINE, MEDLINE, Chemical and Biological Abstract databases. 3 ------- 12-20-04 REVIEW OF PERTINENT LITERATURE Human Studies No reports were located regarding the subchronic or chronic toxicity of any of the alizarin red compounds in humans by inhalation exposure. Animal Studies No reports were located regarding the subchronic or chronic toxicity of any of the alizarin red compounds in animals by inhalation exposure. Other Studies The only toxicity data available for the alizarin red compounds are a few acute studies and case reports. For example, data from acute rabbit studies suggest that alizarin red (CASRN 72-48-0) is a skin sensitizer (Greenburg and Lester, 1954) and a mild eye irritant (500 mg/24 hours) (NIOSH, 2002b). In humans, two case reports of contact dermatitis from the perennial herbs Rubia tinctorum and R. peregrina, both of which contain alizarin red (CASRN 72-48-0) naturally, were published by Castelain and Ducombs (1988). An acute toxicity study in mice is available for alizarin red S (CASRN 130-22-3) wherein an intravenous LD50 of 70 mg/kg was reported (NIOSH, 2002b). No reports regarding acute, subchronic or chronic toxicity in humans or animals by any route were located for any of the other alizarin red compounds. FEASIBILITY OF DERIVING A PROVISIONAL SUBCHRONIC OR CHRONIC RfC FOR ALIZARIN RED COMPOUNDS No pertinent data regarding the inhalation toxicity of alizarin red compounds in humans or experimental animals were located, precluding derivation of subchronic or chronic p-RfCs for alizarin red compounds. REFERENCES ACGIH (American Conference of Governmental Industrial Hygienists). 2001. 2001 Threshold Limit Values for Chemical Substances and Physical Agents and Biological Exposure Indices. Cincinnati, OH. ATSDR (Agency for Toxic Substances and Disease Registry). 2002. Toxicological Profile Information Sheet. Online, http://www.atsdr.cdc.gov/toxpro2.html 4 ------- 12-20-04 Castelain, M. and G. Ducombs. 1988. Contact dermatitis from madder. Con. Derm. 19: 228-229. Greenburg, L.A. and D. Lester. 1954. Handbook of Cosmetic Materials. Interscience, New York. (Cited in Castelain and Ducombs, 1988) IARC (International Agency for Research on Cancer). 2002. Monographs Database. Online. http://www-cie.iarc.fr/ NIOSH (National Institute for Occupational Safety and Health). 2002a. Online NIOSH Pocket Guide to Chemical Hazards. Index of Chemical Abstract Numbers (CAS No.). Online. http ://www. cdc. go v/niosh/npg/npgdcas .html NIOSH (National Institute for Occupational Safety and Health). 2002b. RTECS (Registry of Toxic Effects of Chemical Substances). Cincinnati, OH. NTP (National Toxicology Program). 2002. Management Status Report. Online. http://ntp-server.niehs.nih.gov/ OSHA (Occupational Safety and Health Administration). 2002. OSHA Standard 1910.1000 TableZ-1. Part Z, Toxic and Hazardous Substances. Online. http://www.osha-slc.gov/OshStd data/1910 1000 TABLE Z-l.html 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 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 PB97-921199. 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 Health Criteria series. Online, http://www.inchem.org/ehc.html 5 ------- 12-20-04 Provisional Peer Reviewed Toxicity Values for Alizarin Red Compounds (Various CASRNs) 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 ------- 12-20-04 PROVISIONAL PEER REVIEWED TOXICITY VALUES FOR ALIZARIN RED COMPOUNDS (VARIOUS CASRNS) 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 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 ------- 12-20-04 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 In the chemical literature, several compounds share the common chemical names "alizarin," "alizarin red" or "alizarine red." Seven distinct chemicals were identified in searches of the CAS Registry. Four of the alizarin chemicals are anthraquinone derivatives, two are azo dyes, and one is of an unknown molecular and structural formula. The alizarin red compounds identified include: Anthraquinone derivatives: 1. alizarin, alizarin red, l,2-dihydroxy-9,10-anthracenedione, or l,2-dihydroxy-9,10-anthraquinone (CASRN 72-48-0); 2 ------- 12-20-04 2. alizarin red S, acid red alizarin, [9,10-dihydro-3,4-dihydroxy-9,10-dioxo-2- anthracenesulfonic acid, monosodium salt], or sodium 3,4-dihydroxyanthraquinone-2- sulfonate (CASRN 130-22-3); 3. alizarine red 3 WS or 9,10-dihydro-3,4,7-trihydro-9,10-dioxo-2-anthracenesulfonic acid, monosodium salt (CASRN 6486-94-8); 4. alizarine red YCAP, l,2,6-trihydroxy-9,10-anthracenedione, or 1,2,6-trihydroxyanthraquinone (CASRN 82-29-1); Azo dyes: 5. acid alizarin red B or 2-[(2-hydroxy-3,6-disulfo-l-naphthalenyl)azo]-benzoic acid, trisodium salt (CASRN 1836-22-2); 6. alizarine chrome red G, acid red 97, or 4,4'-bis[(2-hydroxy-l-naphthalenyl)azo]-[l,r- biphenyl]-2,2'-disulfonic acid, disodium salt (CASRN 10169-02-5). Unknown structural and molecular formula: 7. alizarine red V2A or V2A alizarine red (molecular and structural formula unspecified) (CASRN 12794-90-0). Cancer assessments for the alizarin red compounds identified above are not available on IRIS (U.S. EPA, 2002a), the HEAST (U.S. EPA, 1997) or the Drinking Water Standards and Health Advisories list (U.S. EPA, 2002b). The CARA list (U.S. EPA, 1991, 1994) includes no documents for these chemicals. The carcinogenicity of the alizarin red compounds has not been reviewed by IARC (2002), WHO (2002), or ATSDR (2002). The NTP (2002) Management Status Report provided no relevant information. In June 1996, literature searches were conducted from 1965 to May 1996 in TOXLINE, CANCERLIT, MEDLINE, GENETOX, HSDB, EMIC/EMICBACK, DART/ETICBACK, RTECS andTSCATS for relevant studies. In September 2002, update literature searches were conducted in these databases going back to the previous search in 1996. Additional literature search was conducted by NCEA-Cin from October 2003 through May 2004 using TOXLINE, MEDLINE, Chemical and Biological Abstract databases. 3 ------- 12-20-04 REVIEW OF PERTINENT LITERATURE Human Studies No data regarding the possible carcinogenicity of alizarin red compounds in humans by any route of exposure were located. Animal Studies No reports of animal studies examining the carcinogenicity of alizarin red compounds by any route of exposure were located. Other Studies Alizarin red was negative in bacterial tests for mutagenicity using several strains of S. typhimurium (TA98, TA102, TA1535, TA1538) with or without a metabolic activating system (Brown and Brown, 1976; Kawasaki et al., 1992; Matsushima et al., 1986; Tikkanen et al., 1983; Westendorf et al, 1990). In S. typhimurium TA100, alizarin red showed both negative (Brown and Brown, 1976; Brown, 1980) and positive (Tikkanen et al., 1983) results. Alizarin red was mutagenic to S. typhimurium TA1537 (Brown and Brown, 1976; Brown et al., 1977; Brown, 1980; Westendorf et al., 1990). Alizarin red was mutagenic to S. typhimurium TA2637 with or without metabolic activation (Tikkanen et al., 1983). Positive (Brown, 1980) and inconclusive (Fujita et al., 1976) results have been reported for mutagenicity in B. Subtilis. In mammalian test systems, alizarin red was not mutagenic in the V79/HGPRT assay (Westendorf et al., 1990), did not cause chromosomal damage in CHO cells in vitro (Au and Hsu, 1979), and did not enhance transformation of initiated murine fibroblasts (Westendorf et al., 1990; Wolfle et al., 1990). In tests of unscheduled DNA synthesis using primary rat hepatocytes, alizarin red showed both weakly positive (Westendorf et al., 1990) and negative (Kawai et al., 1986; Wolfle et al., 1990) results. Kawai et al. (1986) reported that alizarin red uncoupled and inhibited oxidative phosphorylation in isolated rat liver mitochondria. Using a 32P-postlabeling technique, Poginsky et al. (1991) found no evidence of in vitro DNA adduct formation when alizarin red (740 |imol ) was incubated with murine DNA and rat liver S9 fraction. Similarly, no increase in DNA adduct formation was found in hepatic, renal, duodenal or colon DNA isolated from male Parkes mice fed 10 mg/day alizarin red in cheese spread for 4 days (Poginsky et al., 1991). Alizarin red S did not induce a mutagenic response in S. typhimurium strains TA98, TA100, TA1535, and TA1537 (tested without exogenous metabolic activation) (Brown et al., 1977). In the B. subtilis rec M45 or H17 assay, alizarin red S damaged DNA (Driscoll et al., 1979). No data regarding the genetic toxicity of alizarine red 3WS were located. 4 ------- 12-20-04 No data regarding the genetic toxicity of alizarine red YCAP were located. Brown et al. (1978) reported inconclusive results for alizarin red B in S. typhimurium TA98, TA100, TA1535, TA1537, and TA1538 (with and without exogenous metabolic activation). Alizarine chrome red G was positive in S. typhimurium strains TA98, TA100, TA1535 and TA1537, but only when tested with exogenous metabolic activation (Elliott and Gregory, 1980). No data regarding the genetic toxicity of alizarine red V2A were located. PROVISIONAL WEIGHT-OF-EVIDENCE CLASSIFICATION In accordance with the proposed cancer guidelines (U.S. EPA, 1999), the available data are inadequate for an assessment of human carcinogenic potential for each of the seven alizarin red compounds (alizarin red, CASRN 72-48-0; alizarin red S, CASRN 130-22-3; alizarine red 3WS, CASRN 6486-94-8; alizarine red YCAP, CASRN 82-29-1; acid alizarin red B, CASRN 1836-22-2; alizarine chrome red G, CASRN 10169-02-5; alizarine red V2A CASRN 12794-90-0). QUANTITATIVE ESTIMATES OF CARCINOGENIC RISK Provisional oral slope factors and inhalation unit risk values for the seven alizarin red compounds cannot be derived because human and animal cancer data are lacking. REFERENCES ATSDR (Agency for Toxic Substances and Disease Registry). 2002. Toxicological Profile Information Sheet. Online, http://www.atsdr.cdc.gov/toxpro2.html Au, W. and T.C. Hsu. 1979. Studies on the clastogenic effects of biologic stains and dyes. Environ. Mutagen. 1:27-35. Brown, J.P. 1980. A review of the genetic effects of naturally occurring flavonoids, anthraquinones and related compounds. Mutat. Res. 75: 243-277. 5 ------- 12-20-04 Brown, J.P. and R.J. Brown. 1976. Mutagenesis by 9,10-anthraquinone derivatives and related compounds in Salmonella typhimurium. Mutat. Res. 40: 203-224. Brown, J.P., P.S. Dietrich and R.J. Brown. 1977. Frameshift mutagenicity of certain naturally phenolic compounds in the "Salmonella/microsome" test: activation of anthraquinone and flavonol glycosides by gut bacterial enzymes. Biochem. Soc. Trans. 5: 1489-1492. Brown, J.P., G.W. Roehm and R.J. Brown. 1978. Mutagenicity testing of certified food colors and related azo, xanthene and triphenylmethane dyes with the Salmonella/microsome. Mutat. Res. 56(3): 249-271. Driscoll, R., P.S. Dietrich and J. P. Brown. 1979. Mutagenicity screening of anthraquinone, benzanthrone and triphenylmethane dyes in the Bacillus subtilis rec assay: Comparison with his- reversion in Salmonella typhimurium TA tester strains. Environ. Mutagen. 1(2): 140. (Abstract) Elliott, J. and A.R. Gregory. 1980. Mutagenicity of a series of benzidine congener based dyes. Vet. Human Toxicol. 22(6): 413-417. Fujita, H., A. Mizuo and K. Hiraga. 1976. Mutagenicity of dyes in the microbial system. Tokyo Toritsu Eisei Kenkyusho Nempo (in Japanese). 27(2): 153-158. (Cited in Liefer et al., 1981) IARC (International Agency for Research on Cancer). 2002. Monographs Database. Online. http://www-cie.iarc.fr/ Kawai, K., H. Mori, S. Sugie et. al. 1986. Genetoxicity in the hepatocyte/DNA repair test and toxicity to liver mitochondria of 1-hydroxyanthraquinone and several dihydroxyanthraquinones. Cell. Biol. Toxicol. 2(4): 457-467. Kawasaki, Y., Y. Goda and K. Yoshihira. 1992. The mutagenic constituents of Rubia tinctorum. Chem. Pharm. Bull. 40(6): 1504-1509. Leifer, Z., T. Kada, M. Mandel et al. 1981. An evaluation of tests using DNA repair-deficient bacteria for predicting genotoxicity and carcinogenicity. Mutat. Res. 87: 211-297. Matsushima, T., M. Muramatsu, O. Yagame et al. 1986. Mutagenicity and chemical structure relations of naturally occurring mutagens from plants. Prog. Clin. Biol. Res. 209B: 133-140. NTP (National Toxicology Program). 2002. Management Status Report. Online. http://ntp-server.niehs.nih.gov/ 6 ------- 12-20-04 Poginsky, B., J. Westendorf, B. Blomeke et al. 1991. Evaluation of DNA-binding activity of hydroxyanthraquinones occurring in Rubia tinctorum. Carcinogenesis. 12(7): 1265-1271. Tikkanen, L., T. Matsushima and S. Natori. 1983. Mutagenicity of anthraquinones in the Salmonella preincubation test. Mutat. Res. 116:297-304. 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 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 PB97-921199. U.S. EPA. 1999. Proposed Guidelines for Cancer Risk Assessment. July 1999. Office of Research and Development, National Center for Environmental Assessment, Washington, DC. 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 Westendorf, J., H. Marquardt, B. Poginsky et al. 1990. Genotoxicity of naturally occurring hydroxanthraquinones. Mutat. Res. 240: 1-12. WHO (World Health Organization). 2002. Online catalogs for the Environmental Health Criteria series. Online, http://www.inchem.org/ehc.html Wolfle, D., C. Schmutte, J. Westendorf and H. Marquardt. 1990. Hydroxyanthraquinones as tumor promoters: enhancement of malignant transformation of C3H mouse fibroblasts and growth stimulation of primary rat hepatocytes. Cancer Res. 50: 6540-6544. 7 ------- |