v/EPA United States Environmental Protection Agency EPA Document** 743-D1-5001 August 2015 Office of Chemical Safety and Pollution Prevention TSCA Work Plan Chemical Problem Formulation and Initial Assessment Cyclic Aliphatic Bromides Cluster Flame Retardants Br Br CASRN 25637-99-4 3194-55-6 3194-57-8 NAME Hexabromocyclododecane 1,2,5,6,9,10-Hexabromocyclododecane 1,2,5,6-Tetrabromocyclooctane August 2015 ------- TABLE OF CONTENTS TABLE OF CONTENTS 2 AUTHORS / CONTRIBUTORS / ACKNOWLEDGEMENTS 5 ABBREVIATIONS 6 EXECUTIVE SUMMARY 8 1 INTRODUCTION 11 1.1 SCOPE OF THE ASSESSMENT 12 1.2 REGULATORY AND ASSESSMENT HISTORY 14 2 PROBLEM FORMULATION 16 2.1 PHYSICAL AND CHEMICAL PROPERTIES 16 2.2 PRODUCTION VOLUME AND USES 17 2.2.1 Production Volume 17 2.2.2 Uses 18 2.2.2.1 Use in Expanded Polystyrene Foam (EPS) and Extruded Polystyrene Foam (XPS) 18 2.2.2.2 Use in Textiles 19 2.2.2.2.1 Use in Automotive Textiles 19 2.2.2.2.2 Historic Use in Textiles 20 2.2.2.3 Use in High Impact Polystyrene (HIPS) 20 2.2.2.4 Other Identified Uses 20 2.2.2.5 Summary of All Uses 21 2.2.2.5.1 Summary of CDR Information 21 2.2.2.5.2 Summary of EU Data 21 2.2.3 Future Market Trends 21 2.3 FATE AND TRANSPORT 21 2.4 EXPOSURES 22 2.4.1 Releases to the Environment 22 2.4.2 Presence in the Environment 23 2.4.2.1 Surface Water 23 2.4.2.2 Wastewater 23 2.4.2.3 Sludge 23 2.4.2.4 Soil 23 2.4.2.5 Sediment 24 2.4.2.6 Biota 24 2.4.3 Occupational Exposure 24 2.4.4 General Population Exposure 25 2.4.4.1 Ambient Air 25 2.4.4.2 Drinking Water 25 2 A A3 Fish Consumption 25 2.4.4.4 Biomonitoring 25 2.4.5 Consumer Exposures 26 2.5 HAZARD ENDPOINTS 26 2.5.1 Ecological Hazard 26 2.5.2 Human Health Hazard 27 2.6 RESULTS OF PROBLEM FORMULATION 27 2.6.1 Conceptual Model 27 2.6.2 Analysis Plan 31 2.6.2.1 Workers 31 2.6.2.2 Risks to General Population and Environmental Biota (aquatic, terrestrial and avian) 32 2.6.2.3 Consumers 33 Page 2 of 97 ------- 2.6.3 Sources and Pathways Excluded From Further Assessment 34 2.6.4 Uncertainties and Data Gaps 34 2.6A.I Exposure Assessment 34 2.6.4.1.1 Releases to the Environment 34 2.6.4.1.2 Occupational Exposure 35 2.6.4.1.3 General Population and Consumer Exposure 35 2.6.4.2 Ecological Endpoints 36 2.6.4.3 Human Health Endpoints 36 REFERENCES 37 APPENDICES 61 Appendix A Regulatory and Assessment History 61 Appendix B Uses Supplement Tables 64 Appendix C Exposure Supplement Tables 69 Appendix D Environmental Hazard Study Summaries 74 D-l PERSISTENCE IN ENVIRONMENTAL MEDIA 74 D-l-1 Air and Water 74 D-l-2 Soil, Sediment and Sludge 74 D-l-3 Water and Wastewater 76 D-l-4 Bioaccumulation 76 D-2 TOXICITY TO AQUATIC ORGANISMS 77 D-2-1 Aquatic Plant Toxicity 77 D-2-2 Aquatic Invertebrate Toxicity. 78 D-2-3 Fish Toxicity 80 D-3 TOXICITY TO TERRESTRIAL ORGANISMS 83 D-3-1 Terrestrial Plant Toxicity 83 D-3-2 Soil Invertebrate Toxicity 83 D-3-3 Avian Toxicity 84 Appendix E Human Health Hazard Study Summaries 85 E-l TOXICOKINETICS 85 E-2 ACUTE TOXICITY STUDIES 85 E-3 REPEATED-DOSE TOXICITY STUDIES 86 E-4 REPRODUCTIVE AND DEVELOPMENTAL TOXICITY STUDIES 88 E-5 SKIN IRRITATION AND SENSITIZATION STUDIES 94 E-6 GENOTOXICITY AND CANCER STUDIES 94 LIST OF TABLES Table 2-1: Select Physical-Chemical Properties * 17 Table 2-2: Summary of Use/Exposure Scenarios Considered for Assessment 28 Page 3 of 97 ------- LIST OF APPENDIX TABLES Table_Apx A-l: Regulatory and Assessment History of HBCD 61 Table_Apx B-l: 2012 CDR Production Data (Data Reported for 2011) 64 Table_Apx B-2: Historic IUR and CDR Production Volumes 65 Table_Apx B-3: Summary of 2011 CDR Production Volume and Use Information 65 Table_Apx B-4: Uses of HBCD as Listed in the 2008 EU Risk Assessment 67 Table_Apx C-l: 2012 CDR Data (Data Reported for 2011) for Release Assessment 69 Table_Apx C-2: Compilation of Release Factors and Other Release-Related Information in Various Risk Assessments 70 Table_Apx C-3: Preliminary Values of Input Variables for Calculation of the Range of Release Rates from Manufacturing Sites or the Processing Sites of Each Processing Step 72 Table_Apx D-l: Percent Decrease in Total Initial Radioactivity in Viable Systems and Abiotic Controls During Sludge, Sediment, and Soil Simulation Tests Using 14C-labeled HBCD 75 Table_Apx D-2: Toxicity of HBCD to Aquatic Plants 78 Table_Apx D-3: Toxicity of HBCD to Aquatic Invertebrates 79 Table_Apx D-4: Toxicity of HBCD to Sediment Organisms 80 Table_ApxD-5: Toxicity of HBCD to Fish 82 Table_Apx D-6: Toxicity of HBCD to Terrestrial Plants 83 Table_Apx D-7: Toxicity of HBCD to Terrestrial Invertebrates 83 Table_ApxD-8: Toxicity of HBCD to Avian Species 84 Table_Apx E-l: Acute Oral Toxicity of HBCD 85 Table_Apx E-2: Acute Inhalation Toxicity of HBCD 86 Table_Apx E-3: Repeated-Dose Toxicity of HBCD 87 Table_Apx E-4: Reproductive and Developmental Toxicity of HBCD* 89 Table_Apx E-5: Summary of Effects in Parental and Fl Rats After Dietary, Gestational, Lactational and Postnatal Exposure to HBCD 93 Table_Apx E-6: Genotoxicity of HBCD 96 LIST OF FIGURES Figure 1-1: Generic Structure of Cyclic Aliphatic Bromides 12 Figure 2-1: Conceptual Model for HBCD 28 LIST OF APPENDIX FIGURES Figure_Apx 2.6.4-1: Stepwise Reductive Dehalogenation of HBCD 76 Figure_Apx 2.6.4-2: Exposure-Response Array for Developmental and Reproductive Toxicity Studies of HBCD 92 Figure_Apx 2.6.4-3: Exposure-Response Array for Neurological Effects of HBCD 94 Page 4 of 97 ------- AUTHORS / CONTRIBUTORS /ACKNOWLEDGEMENTS This report was developed by the United States Environmental Protection Agency (US EPA), Office of Chemical Safety and Pollution Prevention (OCSPP), Office of Pollution Prevention and Toxics (OPPT). The Work Plan Chemical Problem Formulation for the cyclic aliphatic bromides cluster was prepared based on currently available data. Mention of trade names does not constitute endorsement by EPA. EPA Assessment Team Leads: Susan A. Laessig, OPPT/Risk Assessment Division (RAD) Maria Szilagyi, OPPT/RAD Team Members: Katherine Anitole, OPPT/RAD Charles Bevington, OPPT/RAD Christina Cinalli, OPPT/RAD Ana Corado, OPPT/Environmental Assistance Division (EAD) Majd El-Zoobi, OPPT/RAD Conrad Flessner, OPPT/RAD Greg Fritz, OPPT/Chemistry, Economics & Sustainable Strategies Division (CESSD) Amuel Kennedy, OPPT/RAD Emma Lavoie, OPPT/CESSD Timothy Lehman, OPPT/CESSD Laurence Libelo, OPPT/RAD Sue Slotnick, OPPT/National Program Chemicals Division (NPCD) Teresa Washington, OPPT/RAD Eva Wong, OPPT/RAD Management Leads: MarkTownsend, OPPT/RAD Nhan Nguyen, OPPT/RAD Acknowledgements Andrea Pavlick, visiting AAAS Science & Technology Policy Fellow, contributed to discussions during problem formulation. Portions of this document were prepared for EPA/OPPT by Abt Associates, the Eastern Research Group (ERG), Inc., the Syracuse Research Corporation (SRC) and Versar. Docket Please visit the public docket (Docket: EPA-HQ-OPPT-2015-0081) to view supporting information. Page 5 of 97 ------- ABBREVIATIONS BFR Brominated Flame Retardant CASRN Chemical Abstract Service Registry Number CBI Confidential Business Information CDR Chemical Data Reporting CEC Commission for Environmental Cooperation CPSC Consumer Product Safety Commission EC European Commission ECHA European Chemicals Agency EFAST Exposure and Fate Assessment Screening Tool EPA Environmental Protection Agency EPS Expanded Polystyrene EU European Union FR Flame Retardant GD Gestation Day GLP Good Laboratory Practices HBCD Hexabromocyclododecane HIPS High Impact Polystyrene HPV High Production Volume HQ Hazard Quotient IRIS Integrated Risk Information System IUR Inventory Update Reporting Rule kg Kilogram(s) KOW OctanohWater partition coefficient Ib Pound LOEL Lowest Observed Effect Level Log KOW Logarithmic Octanol:Water partition coefficient mg Milligram(s) MOE Margin of Exposure ng nanogram NHANES National Health and Nutrition Examination Survey NICNAS National Industrial Chemicals Notification and Assessment Scheme NKRA Not Known or Reasonably Ascertainable NOAEL No-observed-adverse-effect level OCSPP Office of Chemical Safety and Pollution Prevention OECD Organisation for Economic Co-operation and Development OPPT Office of Pollution Prevention and Toxics OSHA Occupational Safety and Health Administration PBT Persistent, Bioaccumulative and Toxic pg picogram PMN Premanufacturing Notice PS Polystyrene Page 6 of 97 ------- PV Production Volume PVC Polyvinylchloride RA Risk Assessment RAR Risk Assessment Report REACH Registration, Evaluation, Authorisation and Restriction of Chemicals SNUR Significant New Use Rule SVOCs Semi-volatile organic chemicals TSCA Toxic Substances Control Act TG Test Guideline US United States WHO World Health Organisation WWTP Wastewater Treatment Plant XPS Extruded polystyrene yr Year Page 7 of 97 ------- EXECUTIVE SUMMARY As part of EPA's comprehensive approach to enhance the Agency's management of existing chemicals, EPA/OPPT identified a work plan of chemicals for further assessment under the Toxic Substances Control Act (TSCA) in March 2012. Chemical risk assessments will be conducted if, as a result of scoping and problem formulation, there are exposures of concern, identified hazards and sufficient data for quantitative analysis. If an assessment identifies unreasonable risks to humans or the environment, EPA will pursue risk reduction. This document presents the problem formulation and initial assessment for the cyclic aliphatic bromides cluster as part of the TSCA Work Plan. EPA/OPPT has identified a cluster of cyclic aliphatic bromide flame retardant chemicals, including, hexabromocyclododecane (HBCD; CASRN 25637-99-4), 1,2,5,6,9,10- hexabromocyclododecane (1,2,5,6,9,10-HBCD; CASRN 3194-55-6) and 1,2,5,6- tetrabromocyclooctane (CASRN 3195-57-8), for risk assessment. These three chemicals have similar physical and chemical properties, and environmental fate characteristics. Uses for 1,2,5,6-tetrabromocyclooctane have not been identified; the remaining two members of the cluster are used as flame retardants in polystyrene foams. HBCD and 1,2,5,6,9,10-HBCD have similar toxicological properties: known effects on the liver and reproductive system. For the purposes of this assessment, the use of "HBCD" refers to either CASRN (25637-99-4 or 3194-55-6), or both. In addition, the conclusions drawn for this assessment will be applicable to both CASRNs. The conclusions from this problem formulation and initial assessment are that EPA/OPPT will evaluate current risk assessments, and if needed, conduct additional analyses as follows: • Workers: Evaluate the applicability of data from published risk assessments to US occupational exposure scenarios to determine if further assessment is needed. If the available data are not applicable, develop estimates of occupational exposures based on modeling and assumptions (e.g. approaches used in the new chemicals program). • General population and biota (aquatic, terrestrial and avian): Estimate releases to the environment in the US to evaluate potential exposure of general population and biota (aquatic, terrestrial and avian) to HBCD. The estimation approach may be based on information in available assessments, coupled with US specific information and/or estimation methods and assumptions. • Consumers: Use available or modeled data relevant to US exposure scenarios to estimate consumer exposure using available or modeled data relevant to US exposure scenarios with particular emphasis on sensitive populations. Several scenarios were identified where exposure to HBCD is expected to be low or unknown and further analysis is not recommended by EPA/OPPT under TSCA: Page 8 of 97 ------- • General population and environmental exposure from HBCD in landfills is not being assessed due to uncertainties in release from these sites. • General population exposure from HBCD in drinking water is not being assessed because drinking water monitoring data for the US are not available and conclusions from available risk assessments indicate a low concern from this exposure pathway (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012). • Consumer exposure to HBCD in High Impact Polystyrene (HIPS) is not being assessed because the level of HBCD in HIPS in the US is unknown, it is not used in typical consumer products (e.g. computer or TV chassis), its use in other consumer products (e.g. electrical appliances) is enclosed limiting potential exposure and a low risk to consumers was indicated in available risk assessments. • Consumer exposure to HBCD in textile finishings is not being assessed because it was considered low risk by the CPSC in upholstered furnishings (CPSC, 2001), it was not reported to be used in consumer fabrics or textiles in the 2012 CDR (EPA, 2012a) and the extent of institutional (e.g. prisons), military or aviation use is unknown. • There are no adequate toxicological data based on inhalation or dermal exposures, nor is there a PBPK model readily available for route-to-route extrapolation. Therefore EPA/OPPT will not assess inhalation or dermal contact in this assessment. However, EPA is considering the quantification of incidental ingestion of particulates that would result from exposure to HBCD dust in occupational settings. A similar approach will be used to address consumer exposure to HBCD in dust. • There are no adequate lifetime exposure or carcinogenicity studies for HBCD. • Inhalation, dermal and lifetime exposure assessments are data gaps that add uncertainty to EPA's risk assessment of HBCD. Hexabromocyclododecane (HBCD) has been used as a flame retardant in plastics (additive) and textiles (backcoating) since the 1980s. Evidence suggests that HBCD is bioaccumulative, environmentally persistent and toxic. Consequently, risk to human health and the environment have been assessed by several countries and global organizations. In 2010, OPPT prepared an action plan for HBCD. Subsequently, HBCD has been nominated for listing on the Toxic Releases Inventory (TRI; in review 2014) and EPA proposed a significant new use rule (SNUR) for use in consumer textiles (EPA, 2012e). During problem formulation, EPA/OPPT identified available fate, exposure and hazard data, and characterized potential exposures, receptors and effects. Data adequacy was determined by Page 9 of 97 ------- following published EPA/OPPT criteria1. EPA/OPPT reviewed the public literature (nominally to September 2014) and Agency information sources (public and confidential) to explore the sources, pathways, receptors and effects for consideration in the assessment. EPA also identified areas of data uncertainty and assumptions. Likely sources and pathways considered for analysis include: use of HBCD as a flame retardant in expanded polystyrene foam (EPS) and extruded polystyrene foam (XPS) in the building and construction industry accounting for 95% of HBCD use mainly in the form of insulation boards. The remaining uses are for high impact polystyrene (HIPS), mainly used for electronics, appliances and possible HBCD-containing textiles for institutional (e.g. prisons), military and aviation uses only (EPA, 2012e). HBCD is not used in consumer textiles that are manufactured in or imported into the US except for limited uses in certain automotive textiles. As outlined in the Conceptual Model for HBCD (Figure 2-1) EPA/OPPT identified the relevant TSCA use of HBCD for this assessment as its use as flame retardants in EPS and XPS products found in commercial and residential environments. EPA/OPPT determined that the major source of exposure to HBCD for human health and the environment was via HBCD dust and/or HBCD in dust generated during the manufacture and processing of HBCD, and the processing and use of products containing HBCD. HBCD in the form of dust or attached to particulates has been measured in indoor domestic and commercial environments, therefore there may be risks to consumers. Preliminary exposure calculations for the US population suggest that the methodology used in available assessments underestimates consumer exposure to HBCD from dust for US consumers. Of particular interest for evaluation are toddlers whose exposure to HBCD from dust in non-residential microenvironments contributes to their total HBCD exposure (Abdallah and Harrad, 2011). HBCD may also make its way into the outdoor environment by transportation through the air and/or washed down the drains (or storm drains) to enter waterways. These exposure scenarios have been considered in risk assessments conducted by other countries and the toxicity and risk of HBCD to aquatic organisms and human health have been assessed and summarized in several publications (EC, 2008; Environment CA and Health CA, 2011; EPA, 2008a, 2014b; NICNAS, 2012; OECD, 2007). However, it is unknown how these conclusions apply to current HBCD manufacture, processing, use and exposure in the US. Therefore, EPA plans to evaluate information in the non-US published risk assessments to determine whether data from other countries are relevant and applicable to US exposures, and where appropriate, supplementing these risk assessments with current and US specific information. 1 Generally followed guidance outlined for the High Production Volume Challenge Program at: http://www.epa.gov/chemrtk/pubs/general/datadfin.htm and http://www.epa.gov/champ/pubs/hpv/Methodology%20for%20HBP%20under%20ChAMP March%202009.pdf and EPA Risk Assessment Guidance at: http://www.epa.gov/raf/ Page 10 of 97 ------- The results of problem formulation as illustrated in the conceptual model and described under the assessment questions indicate that: • There is the potential for occupational exposure to HBCD during HBCD manufacture and processing and polystyrene foam manufacture and processing. • There is potential for general population exposure to HBCD from releases to the environment (air, water, soil and fish consumption). • There is potential for environmental exposure in water, sediment and soil to HBCD from releases to the environment. • There is potential for consumer exposure to HBCD from the use of consumer products in indoor environments. In summary, as a result of problem formulation, EPA/OPPT plans to evaluate current risk assessments and conduct additional risk analysis on potential worker, general population, consumer and environmental exposures under the TSCA Existing Chemicals Program using existing data and methods. EPA/OPPT plans to review and evaluate available exposure (See Section 2.4 and Appendix C) and hazard benchmarks (Section 2.5, Appendix D and Appendix E) and to evaluate the potential non-cancer risk to humans using a margin of exposure approach and potential risks to environment using a hazard quotient approach. As a part of EPA's comprehensive approach to enhance the Agency's management of existing chemicals, in March 2012 EPA/OPPT identified a work plan of chemicals for further assessment under the Toxic Substances Control Act (TSCA)2. After gathering input from stakeholders, EPA/OPPT developed criteria used for identifying chemicals for further assessment3. The criteria focused on chemicals that meet one or more of the following factors: (1) potentially of concern to children's health (for example, because of reproductive or developmental effects); (2) neurotoxic effects; (3) persistent, bioaccumulative, and toxic (PBT); (3) probable or known carcinogens; (4) used in children's products; or (5) detected in biomonitoring programs. Using this methodology, EPA/OPPT identified a TSCA Work Plan of chemicals as candidates for risk assessment in the next several years. In the prioritization process, the Cyclic Aliphatic Bromides Cluster, specifically hexabromocyclododecane (HBCD), was identified for assessment based on its high production volume and PBT characteristics (persistent, bioaccumulative and toxic). EPA/OPPT is performing risk assessments on chemicals in the work plan. If an assessment identifies unacceptable risks to humans or the environment, EPA/OPPT will pursue risk reduction. The target audience for this risk assessment is primarily EPA risk managers; however, it may also be of interest to the broader risk assessment community as well as US stakeholders interested in HBCD. The information presented in the risk assessment may be of assistance to 2 http://www.epa.gov/oppt/existingchemicals/pubs/workplans.html 3 http://www.epa.gov/oppt/existingchemicals/pubs/wpmethods.pdf Page 11 of 97 ------- other federal, state, and local agencies as well as to members of the general public who are interested in the risks of HBCD. The initial step in EPA/OPPT's risk assessment development process, which is distinct from the initial prioritization exercise, includes scoping and problem formulation. During these steps EPA/OPPT reviews currently available data and information, including but not limited to, assessments conducted by others (e.g., authorities in other countries), published or readily available reports and published scientific literature. During scoping and problem formulation the more robust review of the factors influencing initial prioritization may result in refinement • either addition/expansion or removal/contraction - of specific hazard or exposure concerns previously identified in the prioritization methodology. This document includes the results of scoping and problem formulation and initial assessment for HBCD. During problem formulation, EPA/OPPT identified available exposure and hazard data, and characterized potential exposures, receptors and effects. EPA/OPPT developed a conceptual model (Figure 2-1) and analysis plan (Section 2.6.2) as a result of problem formulation. 1.1 Scope of the Assessment The members of the cyclic aliphatic bromides cluster are the brominated flame retardants (BFR): Hexabromocyclododecane (HBCD; CASRN 25637-99-4) 1,2,5,6,9,10-Hexabromocyclododecane (1,2,5,6,9,10-HBCD; CASRN 3194-55-6) 1,2,5,6-Tetrabromocyclooctane (CASRN 3194-57-8)4 EPA prioritized the different BFR chemicals and grouped them into different clusters based on structure. The cyclic aliphatic bromide cluster included chemicals that contain a ring of 6 to 12 saturated carbon atoms with different bromine atoms replacing some of the H atoms on the ring or attached in sidechains. (Br)x Figure 1-1: Generic Structure of Cyclic Aliphatic Bromides Chemicals considered for this cluster were: l,2-Dibromo-4-(l,2-dibromoethyl) cyclohexane [CASRN: 3322-93-8], 1,2,3,4,5,6-hexabromocyclohexane [CASRN: 1837-91-8], 1,2,3,4,5- Pentabromo-6-chlorocyclohexane [CASRN: 87-84-3] and 1,2,5,6-tetrabromocyclooctane [CASRN: 3194-57-8]. One other chemical, Accession number 27248, with generic name polybromocycloalkane would be a member of this Work Plan of chemicals; however, no 4 No domestic uses were identified for 1,2,5,6-Tetrabromocyclooctane (CASRN 3194-57-8). This flame retardant is not functional in current EPS and XPS manufacturing processes. Its thermal stability does not meet operating temperature requirements for the manufacture of XPS foam (EPA, 2014b). Page 12 of 97 ------- production volume has been reported since 1977. Therefore, these chemicals were rejected, along with the individual 1,2,5,6,9,10-HBCD diastereomers, because they were either not on the TSCA Inventory or were not manufactured at sufficient production volume to be reported in the IUR/CDR data collection. Two HBCD commercial chemicals meet this cluster criteria and are the subject of this assessment. These are the only two chemicals being considered for problem formulation in this work plan which differs from the action plan (EPA, 2010a) inclusion criteria. • Hexabromocyclododecane [CASRN 25637-99-4] is produced as a mixture of 16 possible isomers of 1,2,5,6,9,10-hexabromocyclododecane from the bromination of 1,5,9- cyclododecatriene. HBCD is the only member of this cluster that is on the TSCA Inventory that has a significant production volume (as reported in the IUR and CDR). • 1,2,5,6,9,10-hexabromocyclododecane [CASRN 3194-55-6] is a mixture of three main diastereomers of the 16 possible 1,2,5,6,9,10-hexabromocyclododecane isomers. Each individual isomer in this HBCD cluster member contains a ring of 12 saturated carbon atoms with 6 bromine atoms replacing 6 hydrogen atoms. Each isomer has a molecular formula of C12H18Br6. The three most common individual diastereomers are designated as alpha-, beta-, and gamma-HBCD and each has an individual CAS Registry Number (as do all 16 isomers)5. For the purposes of this assessment, the use of "HBCD" refers to either CASRN (25637-99-4 and 3194-55-6), or both. In addition, the conclusions drawn for this assessment will be applicable to both CASRNs. Section 2.6.1 presents the conceptual model developed by EPA/OPPT for HBCD. Using available tools and approaches, the Agency identified the relevant TSCA use of HBCD for this assessment is its use as flame retardants in EPS and XPS products found in commercial and residential environments. Its use in HIPS is not being assessed because the level of HBCD in HIPS in the US is unknown, it is not used in typical consumer products (e.g. computer or TV chassis), its use in other consumer products (e.g. electrical appliances) is enclosed limiting potential exposure and a low risk to consumers was indicated in available risk assessments. Consumer exposure to HBCD in textile finishings is not being assessed because it was considered low risk by the CPSC in upholstered furnishings (CPSC, 2001), it was not reported to be used in consumer fabrics or textiles in the 2012 CDR (EPA, 2012a) and the extent of institutional (e.g. prisons), military or aviation use is unknown (EPA, 2012e). EPA determined that the major source of exposure to HBCD for human health and the environment was via HBCD dust and/or HBCD in dust generated during the manufacture and processing of HBCD, and the processing and use of products containing HBCD. HBCD in the form of dust or attached to particulates has been measured in indoor domestic and commercial 5 This is significant because much of the data are reported for the individual alpha, beta, and gamma isomers rather than for the two commercial products. Page 13 of 97 ------- environments, therefore there may be risks to consumers. Preliminary exposure calculations for the US population suggest that the methodology used in available assessments underestimates consumer exposure to HBCD from dust for US consumers. Of particular interest for evaluation are toddlers whose exposure to HBCD from dust in non-residential microenvironments contributes to their total HBCD exposure (Abdallah and Harrad, 2011). HBCD may also make its way into the outdoor environment by transportation through the air and/or washed down the drains (or storm drains) to enter waterways. These exposure scenarios have been considered in risk assessments conducted by other countries and the toxicity and risk of HBCD to aquatic organisms and human health have been assessed and summarized in several publications (EC, 2008; Environment CA and Health CA, 2011; EPA, 2008a, 2014b; NICNAS, 2012; OECD, 2007). However, it is unknown how these conclusions apply to current HBCD manufacture, processing, use and exposure in the US. Therefore, EPA plans to evaluate information in the non-US published risk assessments to determine whether data from other countries are relevant and applicable to those in the US, and where appropriate, supplement these risk assessments with current and US specific information. 1.2 Regulatory and Assessment History The regulatory and assessment history of HBCD in the US and internationally are summarized in Table_Apx A-l. United States - National HBCD was sponsored in the HPV Challenge Program by BFRIP (BFRIP, 2001). Subsequently, EPA/OPPT prepared a risk based prioritization document in 2008 (EPA, 2008a) which concluded that there was a high concern for potential risk to aquatic organisms, a medium concern for potential risk to the general population from environmental releases and a high concern for potential risk to workers, consumers and children. In 2010, EPA/OPPT prepared an action plan for HBCD (EPA, 2010a). Subsequently, HBCD has been nominated for listing on the Toxic Releases Inventory (TRI; in review 2014) and is subject to rulemaking for use in textiles (EPA, 2012e). In addition, OPPT/DfE (OPPT Design for the Environment) published a flame retardant alternatives assessment for HBCD in 2014 (EPA, 2014). HBCD is currently on the EPA Integrated Risk Information System (IRIS) program agenda. The anticipated date for a completed assessment has not yet been determined (EPA, 2015c). HBCD is not regulated in drinking water under the National Primary Drinking Water Regulations (EPA, 2015b) and is not on the Contaminant Candidate List (CCL) (EPA, 2015a). Published risk assessments indicate low risk to the general population from drinking water exposure (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012). HBCD is under consideration for inclusion in the NHANES human bio-monitoring program. Page 14 of 97 ------- In 2006, the Consumer Product Safety Commission (CPSC) assessed the risk of exposure to HBCD in residential upholstered furniture (CPSC, 2001) and concluded that HBCD did not present a hazard to consumers, as defined by the Federal Hazardous Substances Act (FHSA). The Occupational Safety and Health Administration (OSHA) has not established occupational exposure limits for HBCD. United States-States In California, HBCD is listed as an initial informational candidate under California's Safer Consumer Products regulations (DISC, 2010), on the state's Proposition 65 list (OEHHA, 2007) and is designated a priority chemical for biomonitoring; however, California has not yet started biomonitoring HBCD (SGP, 2014). In Maine, Minnesota and Washington, HBCD is considered a chemical of high concern (DEP, 2013; MDH, 2013; WSDE, 2013). Oregon considers HBCD a priority persistent pollutant (DEQ, 2010a, 2011) and publishes use, exposure pathways and release data for HBCD under this program (DEQ, 2010b). International HBCD is of international concern because of its PBT properties. The toxicity of HBCD to aquatic organisms and human health have been assessed and summarized in several publications (EC, 2008; Environment CA and Health CA, 2011; EPA, 2008a, 2014b; NICNAS, 2012; OECD, 2007). HBCD was added to ECHA's list of Substances of Very High Concern (SVHCs) on October 28, 2008 (ECHA, 2008). Risk assessments have been published by Australia (NICNAS, 2012), Canada (Environment CA and Health CA, 2011) and the European Union (EC, 2008). The conclusions from these assessments are as follows: Occupational, General Population and Consumer Exposure In the Health Canada assessment, the margin of exposure for neurobehavioral effects in infants and children was determined using the LOAEL (0.9 mg/kg-day) from a 90-day study in mice (Eriksson et al., 2006) (Environment CA and Health CA, 2011). This study was not used by the European Commission or the Australian Government (EC, 2008; NICNAS, 2012). For reproductive effects, all three assessments used the NOAEL (10 mg/kg-day) from the two- generation study in rats (Ema et al., 2008). In addition, the European Commission used the NOAEL (22.9 mg/kg-day) from the 28-day study in rats (van der Ven et al., 2006). All three risk assessments concluded that the risk to general population and consumers was of low concern. The occupational risk conclusions vary in different countries due to variations in exposure (e.g. HBCD is not manufactured in Australia) and the risk varies with the activity associated with the extent of exposure to HBCD; low to high, depending on the activity relevant to each country. Environmental Exposure HBCD is persistent and bioaccumulative and is considered a risk to the environment in all three published risk assessments (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012). In May 2013, HBCD was added to the United Nation's Stockholm Convention list of Persistent Page 15 of 97 ------- Organic Pollutants (Stockholm Convention, 2013). The chemical is scheduled to be eliminated by November 2014 with specific exemptions for production and uses in expanded or extruded polystyrene building insulation. As required by the convention, parties that use these exemptions must register with the secretariat and the exemptions will expire in November 2019. Currently, under the Commission for Environmental Cooperation (CEC), Canada, Mexico and the US are evaluating the presence and migration of flame retardants, including HBCD, from consumer products (CEC, 2015). The information gathered from this effort will inform exposure assessors and risk managers and the executive summary of the final report(s) will be available to the public. 2 PROBLEM FORMULATION Problem formulation aims to determine the major factors to be considered in an assessment, including exposure pathways, receptors and health endpoints (EPA, 1998a, 2014c). Accordingly, this problem formulation summarizes the exposure pathways, receptors and health endpoints EPA/OPPT considered to determine whether to conduct further risk analysis and what exposure/hazard scenarios to include in a potential risk assessment. To make this determination, EPA/OPPT conducted a preliminary data review to identify available fate, exposure and hazard data and determine its likely suitability for quantitative analysis and to identify exposure pathways, receptors and health endpoints for quantitative analysis. The outcome of this evaluation is summarized in a conceptual model (Figure 2-1) that illustrates the exposure pathways, receptors and effects that were considered for potential risk assessment. An analysis plan is developed if the results of problem formulation indicate the need for further analysis. 2.1 Physical and Chemical Properties The physical-chemical properties of HBCD are shown in Table 2-1. Commercial preparations of HBCD may contain some impurities, such as tetrabromocyclododecene or other isomeric HBCDs (UNEP, 2010) which are not separately included in this problem formulation. Page 16 of 97 ------- Table 2-1: Select Physical-Chemical Properties * Properties3 Melting Point Boiling Point Vapor Pressure Water Solubility Octanol Water Partition Coefficient (Log Kow) Br Br , ( V^^Br Br -/ <^ Br Br 1,2,5,6,9,10-Hexabromocyclododecane HBCD 175 - 195 °C > 200 °C [decomposes] b 6.27 E-5 Pa at 21°C 66ug/Lat20°C c 5.625 at 25 °C *PCHEM Properties reported in the HPV Robust Summary (BFRIP, 2001) are measured values from a composite of commercial products from 3 different manufacturers. aHPV Data Summary and Test Plan for Hexabromocyclododecane (HBCD) CASRN 3194-55-6 bEC HBCD RAR (EC, 2008) cSum of solubilities for 3 major isomers [alpha, beta, and gamma] in commercial product (ECHA, 2008) 2.2 Production Volume and Uses This section discusses the production volume and uses of the cyclic aliphatic bromides cluster chemicals and is organized as follows: • The 2012 Chemical Data Reporting (CDR) production, import, and export volumes for these chemicals are listed in Appendix B. • Additional details on production volume for HBCD can be found in Section 2.2.1. • Use information can be found in Section 2.2.2. • Future market trends are discussed in Section 2.2.3. 2.2.1 Production Volume EPA/OPPT's 2012 public Chemical Data Reporting (CDR) database (EPA, 2012a), formerly the Inventory Update Reporting (IUR) database includes national-level production volume data; however, the 2012 public CDR database provides limited information on the domestic production volumes of HBCD. For both CAS numbers, site-specific production volumes and national level production volumes were withheld as TSCA Confidential Business Information (CBI) for the 2011 reporting year (EPA, 2012a). Therefore, EPA/OPPT proposes to assess the production volume of HBCD based on the best publicly available production volume data which is the historical IUR and CDR data presented in Appendix B . EPA/OPPT assumes that current Page 17 of 97 ------- production volumes are equal to the most recently reported production volumes (the 2002 and 2006 data for CASRNs 25637-99-4 and 3194-55-6, respectively). For the 2011 reporting year, the data indicate that two sites currently import at least one of the chemicals and that three sites domestically manufacture the chemicals. However, according to the US International Trade Commission, the US imported 92,270 pounds of HBCD (CASRN 25637-99-4) in 2012 (USITC, 2013). This volume does not include HBCD imported as part of an article. Three sites reported export volumes as CBI, and two sites reported no exports (EPA, 2012a). Five sites are identified by the 2012 CDR database as manufacturers or importers of HBCD: BASF Corporation, Albemarle Corporation, The Dow Chemical Company, and two CBI sites (EPA, 2012a). Albemarle manufactures HBCD flame retardants under the Saytex®HP-900 trade name (Albemarle Corporation, 2000). Both Dow and BASF indicate in the CDR data that they are importers; however, trade names of the BASF or Dow Chemical products that use or contain HBCD could not be found in a literature search. For more detailed information on manufacturers of HBCD who reported for the 2012 CDR collection period, see Appendix B. 2.2.2 Uses 2.2.2.1 Use in Expanded Polystyrene Foam (EPS) and Extruded Polystyrene Foam (XPS) HBCD is used as a flame retardant in polystyrene foam, textiles, and high impact polystyrene. The chemical has been in production since the 1960s although there is limited data about the historical use of HBCD in products. The main use of HBCD in the US, the EU, Japan, and Switzerland is as a flame retardant in expanded polystyrene foam (EPS) and extruded polystyrene foam (XPS) (UNEP, 2010; Weil and Levchik, 2009). Use in EPS and XPS accounts for 95 percent of all HBCD applications and began in the 1980s (EPA, 2014b; UNEP, 2010). EPS and XPS are used in the US for thermal insulation boards and laminates for sheathing products used in the building and construction industry. In addition, EPS is used to provide protection from moisture, prevent freezing, provide a stable fill material, and create high-strength composites in construction applications. XPS foam board is used mainly for roofing applications and architectural molding. HBCD is used in both types of foams, because it is highly effective at low-use levels, and therefore maintains the insulation properties of the EPS and XPS foam (Morose, 2006). EPS boards contain approximately 0.5 percent HBCD by weight in the final product while XPS boards contain 0.5 to 1 percent HBCD by weight (Extruded Polystyrene Foam Association, 2011; Morose, 2006). The National Institute of Heath's (NIH) Household Products Database lists HBCD as an ingredient in several extruded and foam insulation products, all of which are manufactured by Owens Corning for use in the US. Currently, Owens Corning lists HBCD in two of its products: Page 18 of 97 ------- Foamular® Extruded Polystyrene Insulation and Foamular® Extruded Polystyrene Insulation - Zero Ozone Depletion Formula, at levels between 0.5 and 1.0 percent (Owens Corning, 2005). The Australian Department of Health and Aging reports that EPS resins are also used in industrial packaging including packaging durable goods and beanbag fill (NICNAS, 2012). Historic data indicate that EPS was used in packaging in North America (Kinshore, 2007), however EPA/OPPT was unable to confirm if this is a current use of HBCD in the US. It should be noted that uses of polystyrene foam in consumer products, such as packaging, generally do not require the use of a flame retardant (EPA, 2014b). 2.2.2.2 Use in Textiles In the US, HBCD was historically used as a flame retardant in the back coating of textiles. However, supported by information gathered from research, industry, and consumer product organizations, EPA/OPPT believes that HBCD is no longer used in consumer textile applications outside of the auto industry. EPA/OPPT received information from a group of textile formulators that the end uses of HBCD-containing textiles are for military, institutional, and aviation applications such as durable carpet tiles for hospitals or prisons (EPA, 2012e; Friddle, 2011). Use in this application is quite small; in 2005 only 1 percent of total production volume of HBCD was used in textiles in the US (EPA, 2012e). HBCD is typically found in textile back coatings at levels of 10-25 percent (Harscher, 2011). In Europe, only 2 percent of HBCD was used in textile applications in 2007 (ECHA, 2009). 2.2.2.2.1 Use in Automotive Textiles Within the US auto industry, EPA/OPPT found that a small amount of HBCD is used in floor mats, headliners, and possibly other interior fabrics in automobiles made or imported to the US (EPA, 2012e). HBCD is currently regulated under Annex XIV of European Union's Regulation on Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), which sets a "sunset date" for the use of the chemical of August 21, 2015. In response to the REACH regulation, the auto industry has formed a consortium to help US manufacturers understand the new requirements; develop tools, processes and best practices; and coordinate compliance efforts. The consortium consists of five North American sponsoring companies, Chrysler, Ford, General Motors, Honda, and Toyota (AIAG, 2011). It is likely that as companies discontinue the use of HBCD in European cars to comply with the REACH regulation, they will discontinue its use in North American automobiles as well. Page 19 of 97 ------- 2.2.2.2.2 Historic Use in Textiles HBCD was historically used as a flame retardant in the US in the back-coating of textiles, upholstered furniture, draperies, wall coverings, and interior textiles such as roller blinds (ECHA, 2009; Morose, 2006). The majority of HBCD used in textiles was for upholstered furniture, because textiles treated with the chemical meet the stringent fire safety laws of the United Kingdom (UK) and California (Morose, 2006). In the 2006 IUR data, one manufacturer/importer of HBCD (CASRN 3194-55-6) reported the use of the chemical substance under the NAICS code for textile and fabric finishing mills (EPA, 2006a). For this use, less than 1 percent of the total production volume of the chemical substance was in consumer and commercial products. The reporting does not distinguish between commercial and consumer use (EPA, 2006a). EPA/OPPT conducted research to determine whether HBCD was used in textile applications for end products sold to consumers in the US. In 2010, an HBCD expert with the Consumer Product Safety Commission (CPSC) expressed to EPA/OPPT his understanding that HBCD is used only in non-consumer textiles such as firefighters' suits (EPA, 2012e). In 2011, EPA/OPPT requested information from current and former manufacturers of HBCD. The responses indicated that only one manufacturer sells HBCD for textile uses. The company did not know whether the end use of any of those textiles is a consumer article (EPA, 2012e). Additionally, a representative of the furniture manufacturing company Herman Miller told EPA/OPPT that HBCD is not in its products (EPA, 2012e). HBCD was not reported to be used in fabrics or textiles in the 2012 CDR (EPA, 2012a). 2.2.2.3 Use in High Impact Polystyrene (HIPS) In both the US and Europe, HBCD is used as a flame retardant in high impact polystyrene (HIPS) for electrical and electronic appliances such as audio-visual equipment, refrigerator lining, and some wire and cable applications (ECHA, 2009; Morose, 2006). Use in television sets is the predominant application of HIPS (Weil and Levchik, 2009). HBCD is found in HIPS products in levels of 1-7% by weight (EC, 2008). Similar data for the US are not available. 2.2.2.4 Other Identified Uses The Australian Department of Health and Aging also reports that minimal amounts of HBCD are imported into the country already incorporated into various articles such as inkjet printers, projectors, scanners, ventilation units for offices, compact fluorescent lights, and LCD digital audiovisual systems (NICNAS, 2012). There are no data to indicate that HBCD is used in the US for these uses. Page 20 of 97 ------- 2.2.2.5 Summary of All Uses 2.2.2.5.1 Summary of CDR Information Appendix B summarizes the HBCD use data as reported in the 2012 CDR. This Appendix also presents information on potential end uses of the chemical beyond what is reported in the CDR. The information is based on additional sources as described in the preceding sections of the report. 2.2.2.5.2 Summary of EU Data Table_Apx B-4 provides a summary of HBCD uses and potential end products as presented in the EU risk assessment report (EC, 2008). Although the EU market and industry for HBCD are considered to be similar to those in the US, differences do exist in building technologies, climate, and consumption patterns, limiting the comparison of the two markets. 2.2.3 Future Market Trends EPA/OPPT expects future use of HBCD to decrease worldwide as the result of forthcoming international regulations. HBCD is listed under Annex XIV of European Union's REACH, which sets a "sunset date" for August 21, 2015. After this date, only persons with approved authorization applications may continue to use the chemical (BSEF, 2012). In addition, in May of 2013, the Conference of the Parties (COP) to the Stockholm Convention on Persistent Organic Pollutants (Convention) decided to list HBCD on the Convention's "elimination" annex, with specific exemptions for production and use for expanded polystyrene and extruded polystyrene in buildings for parties listed in the register of specific exemptions for the substance. The specific exemptions can last up to 5 years and, subject to approval by the COP, can be renewed for a period of up to 5 years. However, the US is not a party to the Stockholm Convention and therefore this action is not applicable to the US. Given that HBCD is going to be phased out for some uses in the majority of the world, including the EU, Canada, Australia, and most of Asia, it is likely that global processors and users of HBCD will work towards phasing out the chemical rather than endure the cost of maintaining a separate supply chain for the US. It is expected that the Stockholm Convention may incentivize US processors, manufacturers, and importers to consider alternatives to HBCD for some applications, which may impact future demand growth for the chemical. 2.3 Fate and Transport The environmental fate of HBCD has been summarized in several publications (EC, 2008; Environment CA and Health CA, 2011; EPA, 2008a, 2014b; NICNAS, 2012; OECD, 2007). A general overview of persistence and bioaccumulation is presented below. Additional details can be found in Appendix D. Page 21 of 97 ------- HBCD is persistent in environmental media. It is expected to be stable to hydrolysis and direct photolysis. Measured aerobic biodegradation half-lives either range up to months, or are greater than months. Anaerobic biodegradation may be more rapid but in anaerobic conditions degradation is also slow with half-lives ranging to months or greater. HBCD is expected to sorb to particulates and sediments and have limited mobility in soil. It is expected to volatilize to some extent from soils and water surfaces. In the atmosphere, HBCD is expected to occur primarily as particulates and may undergo long range transport. It will be removed from the atmosphere by wet or dry deposition, and has an estimated vapor phase half-life of 2.1 days for reaction with hydroxyl radicals. HBCD is highly bioaccumulative with measured fish bioconcentration factor (BCF) values of greater than 18,000. 2.4 Exposures 2.4.1 Releases to the Environment HBCD is manufactured or imported as a powder or pellets (EPA, 2012a) and incorporated into a polymer matrix, including polystyrene foam, as an additive that is incorporated into the matrix (EC, 2008). The life cycle of HBCD includes the manufacture and processing of HBCD followed by the commercial and consumer use, service life, and disposal of products that contain HBCD (EC, 2008; Stockholm Convention, 2010). HBCD is released to the environment throughout the life cycle (EC, 2008; EPA, 2014; Stockholm Convention, 2010). TRI data are not yet available for HBCD, but releases from industrial sites to waste water treatment plants (WWTP), surface water, air and landfill are expected (EC, 2008; Environment Canada, 2011; NICNAS, 2012). HBCD is expected to remain largely immobile in landfills (EPA, 2014) and therefore industrial releases to water and air are of greater interest to EPA/OPPT than industrial releases to landfills. Sawing of EPS or XPS during commercial and consumer use results in release of HBCD (EC, 2008). Emissions of HBCD from EPS and XPS and wear of these products result in release of HBCD during their service life (EC, 2008). The total of releases of HBCD from construction sites to air or surface water from professional use of EPS or XPS is large in comparison with the total releases to each of these media from the manufacture of HBCD or processing of HBCD (to make EPS and XPS) (EC, 2008). However, releases from construction sites are dispersed and therefore are likely to be lower than industrial releases on a per-site basis. Disposal of EPS and XPS may result in releases to the environment as a result of demolition of buildings or material that is left on or in the soil (EPA, 2014); EPA/OPPT believes these releases are likely to be lower than industrial releases on a per-site basis. Manufacturing and processing steps to be assessed are summarized in Appendix C. Page 22 of 97 ------- 2.4.2 Presence in the Environment 2.4.2.1 Surface Water Studies of surface water in the US are limited to a study of suspended sediment from the Detroit River, a highly industrialized area. The maximum measured concentration in suspended sediment was 3.7 u.g/kg dw (Marvin et al., 2006) with similar values measured in China and Sweden (Arnot et al., 2009; He, S. et al., 2013). HBCD was identified in lakes, tributaries and streams in China and the UK with measured concentrations in the ng/L levels (BRE, 2009; Harrad et al., 2009; MOE, 2000, 2005). Measurements of marine water were not found, and these values would be expected to be low. Geographically and temporally distributed monitoring data of this cluster in US surface waters were not found. 2.4.2.2 Waste water HBCD in wastewater influent (dissolved phase) at sewage treatment plants in South Africa and the UK were measured at concentrations of ng/L to <1 u.g/L levels (Chokwe et al., 2012; De Boer et al., 2002). Measurements of the suspended phase of influent from sewage treatment plants in the UK and Netherlands were as high as 3800 u.g/kg dw (De Boer et al 2002, Morris et al., 2004). Measurements of suspended phase in effluent were as high as 18 u.g/kg dw (Morris et al., 2004). Measured concentrations of HBCD in wastewater in the US are not available. 2.4.2.3 Sludge Measurements of HBCD levels in sludge have been made throughout Europe (Covaci et al., 2006; De Boer et al., 2002; Gorga et al., 2013; Guerra et al., 2010; Morris et al., 2004) and Asia (Feng et al., 2012; Hwang et al., 2012) with values ranging from non-detect (detection limit = 4 ng/g) at a WWTPs in Spain (Gorga et al., 2013)(Guerra et al., 2010) to as high as 29 mg/kg dw in industrial sludge from Korea (Hwang et al., 2012). A study in the US of processed sludge from activated sludge-type secondary treatment facility treating domestic & industrial waste (including automotive interior manufacturer) found comparatively high levels of HBCD, on the order of g/kg (La Guardia et al., 2010). Samples analyzed from the EPA 2001 National Sewage Sludge Survey showed approximately 20 ug/kg HBCD (Venkatesan and Halden, 2014). 2.4.2.4 Soil Soil sampling is limited to measurements from Sweden (Arnot et al., 2009), Germany and Belgium (Arnot et al., 2009; Covaci et al., 2006), and throughout Asia (Eguchi et al., 2013; Li et al., 2012b; Wang et al., 2013) with the highest values (u.g/kg dw) found in the soil near a HBCD manufacturing plant in the Laizhou Bay area. Studies with measured levels of HBCD in soils in the US were not available. Page 23 of 97 ------- 2.4.2.5 Sediment Sediment measurements have been made in numerous countries throughout the world including Asia, South America, North America, Europe and South Africa (Al-Odaini et al., 2013; Arnot et al., 2009; Baron et al., 2013; Canton et al., 2008; Covaci et al., 2006; De Boer et al., 2002; de Boer et al., 2004; Feng et al., 2012; Guerra et al., 2012; Harrad et al., 2009; He, M.-J. et al., 2013; Klosterhaus et al., 2012; La Guardia et al., 2012; La Guardia et al., 2013; Li et al., 2013; Li et al., 2012b; Managaki et al., 2012; MOE, 2000, 2005; Morris et al., 2004; Xu et al., 2013; Zhang et al., 2013) with the highest value (>300 mg/kg dw) found at the Yadkin River at the outfall downstream from a textile facility in North Carolina, US (La Guardia et al., 2012). 2.4.2.6 Biota HBCD has been reported in several fresh water and marine species throughout North America. In the US, carp from the Hyco River in Virginia were reported with mean HBCD levels of 4640 u.g/kg lipid weight (Chen et al., 2011). HBCD was also measured in the blubber or liver of various marine mammals: Bottlenose dolphin (Johnson-Restrepo et al., 2008), Bull shark (Johnson- Restrepo et al., 2008), Atlantic Sharpnose shark (Johnson-Restrepo et al., 2008), White Sided dolphin (Peck et al., 2008), and California sea lions (Stapleton et al., 2006) with the highest mean concentration of 130 u.g/kg lipid weight reported in the White Sided dolphin (Peck et al., 2008). Similarly, HBCD has been detected in the blubber or liver of marine species (Budakowski and Tomy, 2003; Muir et al., 2006; Tomy et al., 2009) and in whole or the muscle of fresh water fish in Canada (Law et al., 2006a; Tomy et al., 2008). 2.4.3 Occupational Exposure EPA/OPPT considers inhalation and dermal exposure to be important exposure pathways for workers. Sometimes, the inhalation of air-suspended particulate matter that is subsequently trapped in mucous and moved from the respiratory system to the gastrointestinal tract (EPA, 2011b) is a contributor to aggregate exposures. This will be referred to here as incidental ingestion of inhaled particulates. HBCD is manufactured as a powder at two US sites and is imported as pellets at two other sites. The processing of HBCD for the manufacture of EPS and XPS and the subsequent commercial use of these products by workers is described in EC (2008) and EPA (2014b). Industrial and commercial workers are potentially exposed to HBCD (EC, 2008; Kuo et al., 2014; NICNAS, 2012; Zhang et al., 2012). Exposure monitoring data for workers in the US is not available in the scientific literature. The number of potentially exposed industrial workers in the US is estimated to be less than 2100 (EPA, 2012a). The greatest potential for occupational exposure is expected at industrial sites. Inhalation exposure concentrations of HBCD dust in the form of inhalable particles for workers handling standard grade HBCD powder at sites for the manufacture or processing (for the manufacture of XPS or EPS) of HBCD are in the range of 0.1 to 2.5 mg/m3 (EC, 2008). For dermal exposures, Page 24 of 97 ------- the exposure range is 84 to 840 mg/day of HBCD dust (EC, 2008). Workers who cut EPS or XPS boards (e.g., at construction sites) are potentially exposed to HBCD via inhalation at much lower concentrations of HBCD in air in the form of respirable particles (Kuo et al., 2014; Zhang et al., 2012). 2.4.4 General Population Exposure 2.4.4.1 Ambient Air The concentrations of HBCD are generally higher indoors than outdoors. However, spatial variation is likely with proximity to point sources. Concentrations are generally reported in picograms/m3. Samples have been collected in a wide variety of locations including remote locations in the arctic far removed from sources indicating long-range transport. Some studies characterized vapor and particulate phase of HBCD with HBCD most often reported in the particulate phase (Abdallah et al., 2008b; Alaee et al., 2003; Li et al., 2012a; Takigami et al., 2009a;Tueetal., 2013). 2.4.4.2 Drinking Water Measured concentrations of HBCD in drinking water are limited to one study in the UK (BRE, 2009) where sampling from main water inlet and borehole water indicated concentration of 5- 16 u.g/L and samples of process water from the Netherlands were an order of magnitude lower. Monitoring studies identifying HBCD in drinking water in the US are not readily available. 2.4.4.3 Fish Consumption Measured concentrations of HBCD in fish are reported throughout the world, typically in the u.g/kg range and are expected to vary spatially and temporally (Law et al., 2014). Fewer studies are available in the US and Canada (Arnot et al., 2009; Covaci et al., 2006; Ismail et al., 2009; Klosterhausetal., 2012). 2.4.4.4 Biomonitoring While fewer studies have characterized HBCD levels in humans compared to wildlife, several studies have shown detection in human breast milk, blood, adipose tissue and hair in the US and other countries. HBCD has been detected in breast milk at ng/gram lipid levels. (Arnot et al., 2009; Carignan et al., 2012; Covaci et al., 2006; Croes et al., 2012; Devanathan et al., 2012; Malarvannan et al., 2013; Marvin et al., 2011; Pratt et al., 2013; Shi et al., 2013). Two studies have detected HBCD in adipose tissue (Arnot et al., 2009; Johnson-Restrepo et al., 2008). HBCD has also been detected in human blood at ng/g lipid levels (Arnot et al., 2009; Covaci et al., 2006; de Winter-Sorkina et al., 2006; Kicinski et al., 2012; Kim et al., 2013; WWF, 2004). Page 25 of 97 ------- 2.4.5 Consumer Exposures Consumer exposure to HBCD may include inhalation exposure, dermal exposure through direct skin contact with HBCD on the surface of objects or articles, incidental ingestion of inhaled particulates (see 2.4.3), and incidental ingestion of indoor settled dust via hand-to-mouth behaviors. Based on HBCD's relatively low vapor pressure and relatively high octanol-air partition coefficient, it is likely to preferentially partition to smaller suspended particles in the air and larger settled particles in dust (Blanchard et al., 2014; Law et al., 2014). HBCD has been detected in the dust of residences, commercial buildings, automobiles, and airplanes both in the US and other countries. The available assessments have addressed data relevant to US exposure scenarios up to and including 2011. Concentrations vary widely across different microenvironments and within microenvironments and are generally reported in the nanograms/gram or micrograms/gram range (Abdallah et al., 2008a; Abdallah and Harrad, 2010; AN et al., 2012; Allen et al., 2013a; Allen et al., 2013b; Bjorklund et al., 2012; Covaci et al., 2006; D'Hollander et al., 2010; de Wit et al., 2012; Dodson et al., 2012; Harrad et al., 2010; Johnson et al., 2013; Kalachova et al., 2012; Kopp et al., 2012; Kukucka P*, 2013; Ni and Zeng, 2013; Sahlstrom et al., 2012; Shoeib et al., 2012; Stapleton et al., 2008a; Stapleton et al., 2009; Stapleton et al., 2014; Takigami et al., 2008, 2009a; Thuresson et al., 2012; Tue et al., 2013; van den Eede et al., 2012; Wang et al., 2013). HBCD was detected at nanogram levels in handwipe samples in a recent study (Stapleton et al., 2014). HBCD has also been also detected in indoor air. Concentrations are generally reported in picograms/m3 (Abdallah et al., 2008b; Abdallah and Harrad, 2010; de Wit et al., 2012; Ni and Zeng, 2013; Tue etal., 2013). 2.5 Hazard Endpoints 2.5.1 Ecological Hazard The ecological hazard of HBCD has been summarized in several publications (EC, 2008; Environment CA and Health CA, 2011; EPA, 2008a, 2014b; NICNAS, 2012; OECD, 2007). A general overview is presented below. Additional details and tabulated data summaries can be found in Appendix D. HBCD has been tested for acute and chronic aquatic toxicity, soil organisms, sediment organisms, avian species, and terrestrial plants. EPA/OPPT concludes that HBCD is hazardous to the environment. This conclusion is based on the potential for bioaccumulation (fish bioconcentration factor [BCF]=8,974-18,100) and biomagnification (fish biomagnification factor [BMF]=4.3-9.1), observed acute toxicity values as low as 0.009 mg HBCD/L (72-hour EC5o) in the marine algae, Skeletonema costatum, that indicates high aquatic toxicity to plants, a chronic aquatic toxicity value of 0.0042 mg HBCD/L (maximum acceptable toxicant concentration, MATC) in Daphnia magna that indicates high chronic aquatic invertebrate toxicity, and reduced Page 26 of 97 ------- chick survival in Japanese quails (Coturnix coturnixjaponica) at 15 ppm in diet (2.1 mg HBCD/kg- body weight/day) that indicates high terrestrial toxicity (Drottar and Krueger, 1998, 2000; Law et al., 2006b; MOEJ, 2009; Walsh et al., 1987). 2.5.2 Human Health Hazard The human health hazard of HBCD has been summarized in several publications (EC, 2008; Environment CA and Health CA, 2011; EPA, 2008a, 2014b; NICNAS, 2012; OECD, 2007). A general overview is presented below. Additional details and tabulated data summaries can be found in Appendix E. For humans, there is a potential for oral, inhalation and dermal exposure. Available toxicokinetics data in rodents indicate that HBCD is moderately absorbed via the gastrointestinal tract, metabolized, and distributed to a number of tissues, with preferential distribution and accumulation of unchanged HBCD in fatty tissue. Elimination of HBCD is predominantly via feces (as unchanged parent compound), but is also eliminated in the urine (as secondary metabolites). The acute hazard concern is low via the oral, dermal and inhalation routes. The chronic hazard concern is based on reproductive effects which are described in detail in Appendix E. There is also some evidence of neurodevelopmental toxicity suggestive of hearing impairment, however, it is difficult to determine if the effect is due to developmental exposure to HBCD, a result of repeated-dose exposure, or a combination of the two. Available data suggest that HBCD is not genotoxic. No adequate carcinogenicity studies are available (EPA, 2014b). Existing assessments have also concluded, based on genotoxicity information and one limited lifetime study, that HBCD is not carcinogenic (NICNAS, 2012; TemaNord, 2008) or that further study of carcinogenicity is not warranted (EC, 2008; OECD, 2007). However, the only available dietary study evaluating the carcinogenic potential of HBCD in mice is not considered adequate to draw conclusions regarding carcinogenicity (EC, 2008; Environment CA and Health CA, 2011; EPA, 2014b; OECD, 2007). Given this data gap, EPA's HBCD assessment will not include carcinogenicity assessment. 2.6 Results of Problem Formulation The results of problem formulation are a conceptual model, key assessment questions and an analysis plan for human health and the environment (EPA, 1998a, 2014b). 2.6.1 Conceptual Model During problem formulation, a conceptual model (see Figure 2-1) was developed to identify important sources, pathways, and receptors of exposure (See Sections 2.3 and 2.4). Potential exposures to HBCD (derived from the manufacture, processing and use of HBCD-containing polystyrene products) in homes, offices, the environment, and occupational settings were linked to hazard endpoints in human and non-human receptors. Page 27 of 97 ------- EXPOSURE PATHWAYS Commercial Use of PUT PUF/Products * k Dust from Product Installation &Use Legend Solid lines = Pathway can be quantified Dashed lines = Pathway of unknown significance Shaded boxes = Elements proposed for inclusion in assessment Unshaded boxes = Elements lacking adequate data for assessment Figure 2-1: Conceptual Model for HBCD In the conceptual model, the schematic depicts the pathways (denoted by arrows) of potential exposure to HBCD and HBCD dust generated during the manufacture and processing of HBCD and use of HBCD containing products. The solid lines denote the exposure pathways considered likely and with available exposure and hazard data to assess them. The dashed lines designate pathways which are of unknown significance i.e. uncertain, have limited data or which are not quantifiable. The shaded boxes indicate elements proposed for assessment while the unshaded boxes indicate elements lacking adequate data for assessment. These scenarios are elaborated in Table 2-2. Table 2-2: Summary of Use/Exposure Scenarios Considered for Assessment f 1 USE/EXPOSURE SCENARIO CONSIDERED Worker exposure to HBCD during manufacturing and processing POTENTIAL ROUTE OF EXPOSURE ORAL- Unintended oral exposure via the incidental ingestion of inhaled particles of HBCD and HBCD in dust PROPOSED FOR ASSESSMENT YES RATIONALE/LIMITATIONS/ UNCERTAINTIES Risk identified for HBCD handling in occupational settings in non-US assessments Page 28 of 97 ------- # 2 3 4a 4b 4c USE/EXPOSURE SCENARIO CONSIDERED General population exposure from HBCD resulting from releases to the environment Ecological Receptors Consumer exposure to HBCD from the use of consumer products in indoor environments Consumer exposure to HBCD from the use of consumer products in indoor environments Consumer exposure to HBCD from use of EPS/ XPS commercial products. POTENTIAL ROUTE OF EXPOSURE DERMAL ORAL- Ingestion of HBCD particles ORAL -Fish Consumption ORAL -Drinking Water ORAL -Food other than fish from ambient water INHALATION WATER SEDIMENT SOIL ORAL- Incidental ingestion of inhaled particles of HBCD in dust Hand-to-mouth exposure of HBCD from dust INHALATION DERMAL ORAL INHALATION DERMAL PROPOSED FOR ASSESSMENT NO YES YES NO NO NO YES YES YES YES NO NO RATIONALE/LIMITATIONS/ UNCERTAINTIES Experimental data for reliable route extrapolation from oral to inhalation & dermal routes are not available NoTRI data Low risk in available non-US assessments; confirm low risk using US data Low risk in available non-US assessments; No US drinking water monitoring data Low risk in available non-US assessments; Not regulated under TSCA Low risk in available non-US assessments Available US monitoring data and estimated/modeled releases from industrial sites Available US monitoring data and estimated/modeled releases from industrial sites Available US monitoring data and estimated/modeled releases from industrial sites Data available for oral exposures only Experimental data for reliable route extrapolation from oral to dermal route is not available. Inhalation of neat HBCD potentially released from products is expected to contribute less to overall exposure than the ingestion pathway due to low volatility. Exposure considered insignificant and not assessed in EURAR; Risk to workers low in NICNAS; The HBCD content of these boards 1-5%; EPS/XPS boards may generate dust during cutting during construction, renovations or DIY projects. Consumer dust exposure captured in "1.0" above; Experimental data for reliable route extrapolation from oral to dermal route is not available. Inhalation of neat HBCD potentially released from products is not expected due to low volatility. Page 29 of 97 ------- # 4d 4e USE/EXPOSURE SCENARIO CONSIDERED Consumer exposure to HBCD in specific articles made with high impact polystyrene (HIPS) Consumer exposure to HBCD in textile finishings POTENTIAL ROUTE OF EXPOSURE ORAL INHALATION DERMAL DERMAL PROPOSED FOR ASSESSMENT NO NO RATIONALE/LIMITATIONS/ UNCERTAINTIES Low risk in available non-US assessments; Level of HBCD in HIPS in the US is unknown; Not used in typical consumer products (computer or TV chassis); Use in other consumer products (e.g. electrical appliances) is enclosed limiting potential exposure Low risk in available non-US assessments; Low risk in CPSC study with furnishings; In the 2012 CDR, HBCD was not reported to be used in consumer fabrics or textiles; The extent of HBCD institutional (e.g. prisons), military or aviation use is unknown. EPA developed four key questions from the conceptual model. 1. Are there risks to workers exposed to HBCD during manufacturing and processing of HBCD for the manufacture of EPS and XPS? HBCD has been assessed globally (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012). For human health, the toxicological point of departure (POD; NOAEL for the two- generation toxicity study) used in the published risk assessments (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012) to calculate the margin of exposure (MOE) is based on a study that EPA would consider adequate for the oral route of exposure. No chronic hazard data are available for the dermal and inhalation routes of exposure. To address these exposure scenarios and minimize uncertainty in the risk conclusions, EPA uses physiologically-based pharmacokinetic (PBPK) modeling for route-to-route extrapolation; however, these data are not robust for HBCD. Therefore, EPA/OPPT proposes to evaluate the methodology used in the published risk assessments, confirm the study used for the POD, and in conjunction with EPA's assessment of the exposure of workers in the US (Section 2.6.2), determine if there are risks to workers. 2. Are there risks to the general population from HBCD released to the environment during the lifecycleof HBCD? Releases of HBCD to air, including releases to air during manufacture and processing of EPS and XPS, are expected to partition to particulate matter and deposit in the environment (water and soil) and is expected to be bioaccumulated up the food chain. Therefore, for the general population, exposures to HBCD are expected to occur indirectly through water or fish consumption. Drinking Water: No monitoring data for HBCD in drinking water are available. However, published risk assessments indicate low risk to the general population from drinking water exposure. Page 30 of 97 ------- Fish Consumption: • For the general population, available risk assessments (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012) concluded that risk to general population from consumption of HBCD in fish is low. In addition, the Canadian assessment included sensitive subpopulations such as indigenous populations (Nunavut) and nursing infants and concluded that risks to these populations is low. • However, recent publications (Abdallah and Harrad, 2011; Aylward and Hays, 2011; Carignan et al., 2012; Gheorghe et al., 2013; Kalachova et al., 2012) containing additional information warrant evaluation. A biomonitoring-based risk assessment (Aylward and Hays, 2011) (i.e., based on HBCD concentrations found in breast milk and serum) indicated that the margins of exposure (MOE) were greatly in excess of target values, suggesting that the risk to the general population is low. 3. Are there risks to ecological receptors from HBCD found in the environment? Available risk assessments (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012) concluded that HBCD poses a risk to the environment. EPA/OPPT plans to evaluate the potential risk to the environment based on US release estimates and exposure. 4. Are there risks to consumers from HBCD found in household dust? Dust: Available risk assessments (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012) concluded that risk to consumers from exposure to HBCD in household dust is low. These assessments included hand-to-mouth transfer of dust by children. Preliminary evaluation of recent data for the US population suggest that the available assessments underestimate consumer exposure to HBCD from dust for US consumers. In addition, for toddlers, exposure to HBCD from dust in other microenvironments, such as vehicles and childcare environments, may contribute to their total HBCD exposure (Harrad and Abdallah, 2011). Therefore, EPA/OPPT plans to evaluate the methodology used in the published risk assessments, confirm the study used for the hazard assessment and in conjunction with the current and aggregate exposure information relevant to the US population, assess potential risks to US consumers. 2.6.2 Analysis Plan Based on problem formulation EPA/OPPT plans to conduct the following additional analyses. 2.6.2.1 Workers EPA/OPPT plans to evaluate the applicability of data for worker exposure to HBCD through manufacturing and processing for the manufacture of EPS and XPS reported in EC (2008) and NICNAS (2012) to US occupational exposure scenarios. If the available data are not applicable, develop estimates of occupational exposures based on modeling and assumptions (e.g. approaches used in the new chemicals program). Page 31 of 97 ------- 2.6.2.2 Risks to General Population and Environmental Biota (aquatic, terrestrial and avian) Robust monitoring datasets for US locations do not exist. EPA/OPPT plans to use estimates of releases to the environment during HBCD manufacture and processing for the manufacture of EPS and XPS from manufacturing, processing and use to estimate surface water, sediment and soil concentrations. EPA/OPPT does not plan to consider degradation losses but may consider partitioning. EPA/OPPT plans to estimate releases to the environment from industrial sites based on CDR (EPA, 2012a) data on production volume and number of sites and emission factors (i.e. 'loss factors')6 reported in various HBCD risk assessment reports (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012). EPA/OPPT will assume emission factors for releases from manufacturing and processing, in processes in other countries, are applicable to the US. The release factor of a chemical is dependent on the design and operation of a chemical process. EPA/OPPT assumes the basic process design and operation of the processes for the manufacture or processing of HBCD in the US to be similar to those of the corresponding processes in other countries. The descriptions of the processes for the manufacture of EPS resin beads, the manufacture of foamed plastics including EPS and XPS, and plastics compounding in general that are reported in the literature (Burkhardt et al., 2011; EPA, 2014b; Maul et al., 2007; Suh, 2000) are similar to descriptions reported in EC (2008) and NICNAS (2012) of the corresponding processes. EPA/OPPT's compilation of the non-site specific emission factors reported in EC (2008), NICNAS (2012) and Environment CA and Health CA (2011) is given in Table_Apx C-2 in Appendix C. EPA/OPPT's preliminary estimate of the values of input variables for the assessment of releases is reported in Table_Apx C-3; EPA/OPPT plans to assess a range of release values from the ranges of input variables given in this table. Additionally, EPA/OPPT may consider other available information to assess releases and concentrations to other media (i.e. air). For a discussion of the approach for how air releases could be modeled to estimate concentrations in nearby media see EPA/OPPT's TBBPA problem formulation document (http://www.epa.gov/oppt/existingchemicals/pubs/workplans.html). However, there will be important differences in the assessments because site-specific modeling parameters were used with TBBPA and those are not likely to be available for HBCD industrial sites. Mathematical modeling approaches may be necessary to yield exposure estimates. EPA/OPPT will consider the use of sensitivity analyses to determine key elements of uncertainty. EPA/OPPT plans to estimate MOE for fish consumption (including sensitive populations) using US exposure information not captured in previous assessments and modeled fish ingestion (EPA, 2007) from release estimates. EPA/OPPT is considering values for adult general population consumption typically used by EPA Office of Water (e.g., 22 g/day for adults in the 6 Ratio of amount of chemical released to amount manufactured or processed Page 32 of 97 ------- general population and 142.5g/day for subsistence fishers in the absence of local or similar fish ingestion data). Based on NHANES data from 2003 to 2010 (EPA, 2014a), this value represents the 90th percentile consumption rate of freshwater and estuarine fish for the US adult population 21 years of age and older. EPA/OPPT will identify hazard endpoints and benchmarks from published assessments (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012) and data sources (See Appendix D and Appendix E). EPA/OPPT will calculate non-cancer risks using MOE or HQ approaches. 2.6.2.3 Consumers Mathematical modeling approaches may be necessary to yield exposure estimates. EPA/OPPT will consider the use of sensitivity analyses to determine key elements of uncertainty. EPA/OPPT plans to use available or modeled (to be determined) data relevant to US exposure scenarios to estimate consumer exposure using available or modeled data relevant to US exposure scenarios with particular emphasis on sensitive populations (e.g. toddlers exposed in microenvironments). Consumer exposures to HBCD will be evaluated based on incidental ingestion of dust (as described above), and incidental ingestion of indoor settled dust via hand-to-mouth behaviors. Oral exposure by incidental ingestion of house dust and hand-to-mouth transfer can be quantified based on US values of monitored house dust. Several recent studies of house dust are available which are expected to be representative of US households. The EPA Exposure Factors Handbook (EPA, 2011b) can be utilized to determine typical quantities of dust ingested and time-activity patterns. EPA/OPPT will identify hazard endpoints and benchmarks from published assessments (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012) and data sources (See Appendix E). EPA/OPPT will calculate non-cancer risks using MOE. Aggregate oral exposures will be assessed considering hand-to-mouth dust ingestion, incidental ingestion of dust and high-end fish consumption. Conclusion EPA/OPPT plans to evaluate potential risk to workers, the general population, consumers and environmental biota under the TSCA Existing Chemicals Program using existing data and methods. EPA/OPPT plans to review and evaluate available exposure and hazard benchmarks and determine margins of exposure to evaluate the potential risk from human and environmental exposure to HBCD. EPA/OPPT plans to estimate releases to the environment from industrial sites based on CDR (EPA, 2012a) data on production volume and number of sites and emission factors (i.e. 'loss factors')7 reported in various HBCD risk assessment reports (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012). 7 Ratio of amount of chemical released to amount manufactured or processed Page 33 of 97 ------- 2.6.3 Sources and Pathways Excluded From Further Assessment Several scenarios were identified where exposure to HBCD is expected to be low or unknown, and where further analysis is not recommended by EPA/OPPT under TSCA: • Exposure from HBCD in landfills is not being assessed due to uncertainties in release from these sites. • General population exposure from HBCD in drinking water is not being assessed because drinking water monitoring data for the US are not available and conclusions from available risk assessments indicate a low concern from this exposure pathway (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012). • Consumer exposure to HBCD in HIPS is not being assessed because the level of HBCD in HIPS in the US is unknown, it is not used in typical consumer products (e.g. computer or TV chassis), its use in other consumer products (e.g. electrical appliances) is enclosed limiting potential exposure and a low risk to consumers was indicated in available risk assessments. • Consumer exposure to HBCD in textile finishings is not being assessed because it was considered low risk by the CPSC in upholstered furnishings (CPSC, 2001), it was not reported to be used in consumer fabrics or textiles in the 2012 CDR (EPA, 2012a) and the extent of institutional (e.g. prisons), military or aviation use is unknown. • There are no adequate toxicological data based on inhalation or dermal exposures or a PBPK model readily available for route-to-route extrapolation. Therefore EPA/OPPT will not assess inhalation or dermal contact in this assessment. However, EPA is considering the quantification of incidental ingestion of particulates that would result from exposure to HBCD dust in occupational settings. A similar approach will be used to address consumer exposure to HBCD in dust. 2.6.4 Uncertainties and Data Gaps 2.6.4.1 Exposure Assessment 2.6.4.1.1 Releases to the Environment The major uncertainties in EPA's proposed approach are the following: Production and Processing Volumes and Number of Sites: HBCD production and processing volumes are uncertain because CDR information on current production, export and import volumes is CBI. EPA will assume the HBCD production volume to be in a range of values that is derived from the most recent publically reported CDR information Page 34 of 97 ------- and will assume the processing volume to be equal to the production volume. Refer to Table_Apx C-3 for EPA's preliminary values for production and processing volumes. The number of sites for most processing steps is uncertain. EPA will estimate the number of sites based on CDR data which is data on ranges of number of sites. Furthermore, the processing and product descriptions reported in CDR are general and preclude an accurate determination of specific processing steps. Refer to Table_Apx C-l for EPA's preliminary assessment of the number of sites. The share of the HBCD production volume that is processed to manufacture XPS using HBCD powder or HBCD masterbatch is unknown. 2.6.4.1.2 Occupational Exposure Exposure monitoring data for workers in the US are not available in the scientific literature. The greatest potential for occupational exposure is expected at industrial sites. Maximum exposure occurs while workers load or unload HBCD powder or pellets (EC, 2008), which is a worker activity that EPA expects in the US. Data are available for inhalation and dermal exposures to workers (EC, 2008). Workers who cut EPS or XPS boards (e.g., at construction sites) are potentially exposed to HBCD via inhalation at much lower concentrations of HBCD in air in the form of respirable particles. There is no PBPK model readily available for route-to-route extrapolation. EPA/OPPT has identified this as a critical data gap since the exclusion of dermal and inhalation exposure routes will result in the underestimation of risks. 2.6.4.1.3 General Population and Consumer Exposure Some of the available measured environmental concentrations were outside the US and it is not clear how representative they are of exposure scenarios within the US. Significant uncertainties may exist in a quantitative evaluation. There are limited US surface water, sediment, and soil measurements. Available monitoring data may not be representative of concentrations in the environment across all areas of the US. There are very limited data of HBCD in fish from the US and it is uncertain if concentrations in fish in Canada and abroad would be similar to the US. Fish ingestion exposures will need to be modeled based on releases to the environment from manufacturing/processing/use. Exposure factors exist for fish consumption, however there would be uncertainty in determining the concentration of HBCD in edible fish. If specific receiving waters are not identified, there will be uncertainty in the amount of dilution that may occur. EPA/OPPT will clearly document the uncertainty and limitations associated with the fish consumption analyses. Modeled releases to water from industrial facilities may result in the over- or under-estimation of concentrations in the aquatic environment. Modeling default values will need to be modified Page 35 of 97 ------- (e.g., fish consumption) to account for high-end consumption. EPA/OPPT will consider the use of sensitivity analyses to determine key elements of uncertainty. The concentration of HBCD in indoor air or dust in offices or workplaces may be greater than in homes. There are uncertainties using existing methodologies to estimate exposure for the different sub-environments. Incidental ingestion of dust by adults is expected to be low whereas ingestion of dust through incidental ingestion or hand to mouth behavior is expected to be higher for small children due to their activity patterns and increased proximity to indoor areas where dust may gather. Concentrations of HBCD in dust are likely to vary by microenvironment and will need to be a consideration in exposure estimations. It is not possible to develop source-to-dose exposure models with currently available information. Sources such as dust and presence in fish must be considered integrative metrics for the purposes of exposure assessment. Source-to-dose models are absent or limited for most of the identified exposure scenarios, therefore linking the exposure to specific products or the use patterns of any one product will be challenging. 2.6.4.2 Ecological Endpoints Overall, adequate aquatic toxicity data are available to characterize the hazard to the environment for HBCD. 2.6.4.3 Human Health Endpoints Toxicokinetics (by the oral route), acute, repeated-dose, developmental, and reproductive toxicity data are available to characterize the potential human health hazard of HBCD. Although no standard neurotoxicity or developmental neurotoxicity studies on HBCD are available, information on neurotoxicity was obtained from Functional Observational Battery, locomotor activity evaluations, neurobehavioral testing, surface righting reflex, negative geotaxis reflex, mid-air righting reflex, and brainstem auditory evoked potentials (BAEPs) in several repeated-dose and reproductive toxicity studies. Several assays testing for genotoxicity, and irritation/sensitization are also available. The only available dietary study evaluating the carcinogenic potential of HBCD in mice is not considered adequate to draw conclusions regarding carcinogenicity (EC, 2008; Environment CA and Health CA, 2011; EPA, 2014b; OECD, 2007). EPA agrees with these conclusions and therefore in its HBCD assessment will not consider carcinogenicity to be an endpoint of concern. Neither a complete mechanistic PBPK model for HBCD, nor a PBPK model for humans is available. This precludes the use of a model for cross-route or cross-species extrapolation. There is no PBPK model readily available for route-to-route extrapolation. Exclusion of dermal and inhalation exposure routes will result in the underestimation of risks. The lack of a lifetime exposure study and/or adequate assessment of carcinogenicity increases uncertainty in EPA's assessment of long-term exposures to HBCD. Therefore, inhalation, dermal and lifetime exposure assessment data gaps add uncertainty to EPA's risk assessment of HBCD. Page 36 of 97 ------- REFERENCES Abdallah, M., and S. Harrad. 2011. Tetrabromobisphenol-a, Hexabromocyclododecane and Its Degradation Products in UK Human Milk: Relationship to External Exposure. Environment International, 37(1), 443-448. Abdallah, M. A.-E., S. Harrad, and A. Covaci. 2008b. Hexabromocyclododecanes and Tetrabromobisphenol-a in Indoor Air and Dust in Birmingham, UK: Implications for Human Exposure. Environmental Science &Technology, 42(18), 6855-6861. Abdallah, M. A.-E., S. Harrad, C. Ibarra, M. Diamond, L. Melymuk, M. Robson, and A. Covaci. 2008a. Hexabromocyclododecanes in Indoor Dust from Canada, the United Kingdom, and the United States. Environmental Science &Technology, 42(2), 459-464. Abdallah, M. A. E., and S. Harrad. 2010. Modification and Calibration of a Passive Air Sampler for Monitoring Vapor and Particulate Phase Brominated Flame Retardants in Indoor Air: Application to Car Interiors. Environmental Science & Technology, 44, 3059-3065. AIAG (Automotive Industry Action Group). 2011. Reach Activities. http://www.aiag.org/staticcontent/committees/workgroup.cfm?FC=ES&grp=WCOC&gr oup=OHRE (accessed on March 15, 2011). Al-Odaini, N. A., U. H. Yim, N. S. Kim, W. J. Shim, and S. H. Hong. 2013. Isotopic Dilution Determination of Emerging Flame Retardants in Marine Sediments by Hplc-Apci-Ms/Ms. Analytical Methods, 5,1771-1778. Alaee, M., D. Muir, C. Cannon, P. A. Helm, T. Harner, and T. Bidleman. 2003. New Persistent Chemicals in Arctic Air and Water. In Bidleman, T., R. Macdonald , and J. Stow, Northern Contaminants Program Canadian Arctic Contaminants Assessment Report II: Sources, Occurrence, Trends and Pathways in the Physical Environment (pp. 116-124). Minister of Indian Affairs and Northern Development, Ottawa, CANADA. Albemarle Corporation. 2000. Saytex HP-900 Flame Retardant. Baton Rouge, LA. http://www.albemarle.com/Products---Markets/SAYTEX-HP-900P-AND-HP-900G- 163C18.html. AN, N., A. C. Dirtu, N. Van den Eede, E. Goosey, S. Harrad, H. Neels, A. Mannetje, J. Coackley, J. Douwes, and A. Covaci. 2012. Occurrence of Alternative Flame Retardants in Indoor Dust from New Zealand: Indorr Sources and Human Exposure Assessment. Chemosphere, 88, 1276-1282. Page 37 of 97 ------- Allen, J. G., H. M. Stapleton, J. Vallarino, E. McNeely, M. D. McClean, S. J. Harrad, C. B. Rauert, and J. D. Spengler. 2013a. Exposure to Flame Retardant Chemicals on Commercial Airplanes. Environmental Health, 12,17. Allen, J. G., A. L Sumner, M. G. Nishioka, J. Vallarino, D. J. Turner, H. K. Saltman, and J. D. Spengler. 2013b. Air Concentrations of PBDEs on in-Flight Airplanes and Assessment of Flight Crew Inhalation Exposure. Journal of Exposure Science and Environmental Epidemiology, 23, 337-342. Antignac, J. P., R. Cariou, D. Maume, and et al. 2008. Exposure Assessment of Fetus and Newborn to Brominated Flame Retardants in France: Preliminary Data. Mol Nutr Food Res, 52(2), 258-265. Arita, R., K. Miyazaki, and S. Mure. 1983. Metabolic TestofHBCD. Test on Chemical Substances Used in Household Items. Studies on Pharmacodynamics of H BCD In: Toxicology summary: HBCD (HBCD), Albemarle, S.A. Department of Pharmacy, Hokkaido University Hospital, Hokkaido, Japan. Arnot, J., L. McCarty, J. Armitage, L. Toose-Reid, F. Wania, and I. Cousins (European Brominated Flame Retardant Industry Panel (EBFRIP)). 2009. An Evaluation of Hexabromocyclododecane (HBCD) for Persistent Organic Pollutant (POP) Properties and the Potential for Adverse Effects in the Environment. May 26, 2009. Brussels, BE. http://www.unece.org/fileadmin/DAM/env/documents/2009/EB/wg5/wgsr45/lnformal %20docs/An%20evaluation%20of%20hexabromocyclododecane Final%20report.pdf. Aufderheide, J., A. Jones, J. A. MacGregor, and W. B. Nixon. 2003. Effect of Hexabromocyclododecane on the Survival and Reproduction of the Earthworm, Eisenia Fetida. ABC Laboratories, Inc and Wildlife International Ltd, Columbia, MO and Easton, MD. (as cited in BFRIP, 2005). Aylward, L. L., and S. M. Hays. 2011. Biomonitoring-Based Risk Assessment for Hexabromocyclododecane (HBCD). International Journal of Hygiene and Environmental Health, 214(1), 179-187. Baron, E., P. Gago-Ferrero, M. Gorga, I. Rudolph, G. Mendoza, A. M. Zapata, S. Diaz-Cruz, R. Barra, W. Ocampo-Duque, M. Paez, R. M. Darbra, E. Eljarrat, and D. Barcelo. 2013. Occurence of Hydro phobic Organic Pollutants (BFRs and UV-Filters) in Sediments from South America. Chemosphere, 92, 309-316. BASF. 1990. Report on the Study of the Acute Oral Toxicity of HBCD in the Mouse Cover letter dated 031290. Submitted under TSCA Section 8D; EPA Document No. 86-900000383; NTIS No. OTS0522946., Washington, DC. Page 38 of 97 ------- BASF (BASF Aktiengesellschaft). 2000. Cytogenetic Study in Vivo with ofHBCD in the Mouse Micronucleus Test after Two Intraperitoneal Administrations. Project No. 26MO100/004018. Experimental Toxicology and Ecology, Ludwigshafen, Germany. Becher, G. 2005. The Stereochemistry ofl,2,5,6,9,10-Hexabromocyclododecane and Its Graphic Representation. Chemosphere, 58(1), 989-991. BFRIP (American Chemistry Council, Brominated Flame Retardant Industry Panel). 2001. High Production Volume (HPV) Challenge Program Submission for Hexabromocyclododecane (HBCD). Washington, DC. http://www.epa.gov/oppt/chemrtk/pubs/summaries/cyclodod/cl3459tc.htm. Bidleman, T. F. 1988. Atmospheric Processes. Environmental Science & Technology, 22(4), 361- 367. Bjorklund, J. A., U. Sellstrom, C. A. de Wit, M. Aune, S. Lignell, and P. Darnerud. 2012. Comparisons of Polybrominated Diphenyl Ether and Hexabromocyclododecane Concentrations in Dust Collected with Two Sampling Methods and Matched Breast Milk Samples. Indoor Air, 22, 279-288. Blanchard, O., P. Glorennec, F. Mercier, N. Bonvallot, C. Chev, O. Rmalho, C. Mandin, and B. Le Bot. 2014. Semivolatile Organic Compuonds in Indoor Air and Settled Dust 30 French Dwellings. Environmental Science & Technology, 48, 3959-3969. Brandsma, S. H., L. T. Van der Ven, J. De Boer, and P. E. Leonards. 2009. Identification of Hydroxylated Metabolites of Hexabromocyclododecane in Wildlife and 28-Days Exposed Wistar Rats. Environmental Science &Technology, 43(1), 6058-6063. BRE (Building Research Establishment). 2009. Brominated Flame Retardants - Risks to UK Drinking Water Sources. Report number 242844. Garston, UK. Budakowski, W., and G. Tomy. 2003. Congener-Specific Analysis of Hexabromocyclododecane by High-Performance Liquid Chromatography/Electrospray Tandem Mass Spectrometry. Rapid Communications in Mass Spectrometry, 17,1399-1404. Burkhardt, G., U. Husgen, M. Kalwa, G. Potsch, and C. Schwenzer. 2011. Plastics Processing, 1. Processing of Thermoplastics. In Bellussi, G. e. a., Ullmann's Encyclopedia of Industrial Chemistry (Vol. 28, pp. 155-193). Wiley-VCH Verlag GmbH & Co. KGaA, Aachen, Germany, http://dx.doi.ore/10.1002/14356007.a20 663.pub2. Calmbacher, C. W. 1978. The Acute Toxicity ofHBCD Lot 990-17 to the Bluegill Sunfish Lepomis Macrochirus Rafinisque with Test Data and Cover Letter. UCES proj. No. 11506-03-, 9 pp. TSCA Section 8D; OTS0523260 , DCN 86900000268. Union Carbide Corporation, Tarrytown, NY. Page 39 of 97 ------- Canton, R. F., A. A. C. M. Peijnenburg, R. L. A. P. Hoogenboom, A. H. Piersma, L. T. M. van der Ven, M. van den Berg, and M. Heneweer. 2008. Subacute Effects of Hexabromocyclododecane (HBCD) on Hepatic Gene Expression Profiles in Rats. Toxicology and Applied Pharmacology, 231, 267-272. Carignan, C. C., M. A.-E. Abdallah, N. Wu, W. Heiger-Bernays, M. D. McLean, S. Harrad, and T. F. Webster. 2012. Predictors ofTetrabromobisphenol-a (TBBP-A) and Hexabromocyclododecanes (HBCD) in Milk from Boston Mothers. Environmental Science &Technology, 46, 12146-12153. CEC (Commision for Environmental Cooperation). 2015. Enhancing Trilateral Understanding of Chemicals in Products in North America. Montreal, Canada. http://www.cec.ore/Paee.asp?PagelD=122&ContentlD=25630&SiteNodelD=1299&BL E xpandID (accessed on May 12, 2015). Chen, D., M. J. La Guardia, D. R. Luellen, E. Harvey, T. M. Mainor, and R. C. Hale. 2011. Do Temporal and Geographical Patterns of HBCD and PBDE Flame Retardants in the U.S. Fish Reflect Evolving Industrial Usage? Environmental Science & Technology, 45(19), 8254-8261. Chengelis, C. 2002. Amendment to the Final Report For: An Oral (Gavage) 90-Day Toxicity Study of HBCD in Rats. Study No. WIL-186012.. WIL Research Laboratories, Inc, Ashland, OH. http://www.epa.gov/oppt/tsca8e/pubs/8ehq/2002/sep02/fyi 0902 01424b2.pdf (Accessed January 14, 2014). Chengelis, C. P. 1997. A 28-Day Repeated Dose Oral Toxicity Study of HBCD in Rats Document No. FYI-OTS-0397-1289. WIL Research Laboratory, Inc. for Chemical Manufacturers Association, Ashland, OH. Chengelis, C. P. 2001. An Oral (Gavage) 90-Day Toxicity Study of HBCD in Rats. WIL-186012. WIL Research Laboratory, Inc. for Chemical Manufacturers Association, Ashland, OH. Chokwe, T. B., J. O. JOkonkwo, L. L. Sibali, and E. J. Neube. 2012. Optimization and Simultaneous Determination ofAlkyl Phenol Ethoxylates and Brominated Flame Retardants in Water after Spe and Heptafluorobutyric Anhydride Derivatization Followed by Gc/Ms. Chromatographia, 75,1165-1176. CMA (Chemical Manufacturers Association). 1997. Hexabromocyclododecane (HBCD): Determination of the Vapor Pressure Using a Spinning Rotor Gauge. Study conducted by Authors of the Study (Required field. Type "author's last name, first name initials", Wildlife International Ltd,, (July 23, 1997), Easton, MD. OTS0573702. Page 40 of 97 ------- Covaci, A., A. C. Gerecke, R. J. Law, S. Voorspoels, M. Kohler, N. V. Heeb, H. Leslie, C. R. Allchin, and B. J. De. 2006. Hexabromocyclododecanes (HBCDs) in the Environment and Humans: A Review. Environmental Science and Technology, 40(12), 3679-3688. CPSC (US Consumer Product Safety Commission). 2001. CPSC Staff Exposure and Risk Assessment of Flame Retardant Chemicals in Residential Upholstered Furniture. Bethesda, MD. Croes, K., A. Colles, G. Koppen, E. Govarts, L. Bruckers, E. Van de Mieroop, V. Nelen, A. Covaci, A. C. Dirtu, C. Thomsen, L. S. Haug, G. Becher, M. mampaey, G. Schoeters, N. Van Larebeke, and W. Baeyens. 2012. Persistent Organic Pollutants (POPs) in Human Milk: A Biomonitoring Study in Rural Areas of Flanders (Belgium). Chemosphere, 89, 988-994. D'Hollander, W., L. Roosens, A. Covaci, C. Cornelis, H. Reynders, K. Van Campenhout, P. de Voogt, and L. Bervoets. 2010. Brominated Flame Retardants and Perfluorinated Compounds in Indoor Dust from Homes and Offices in Flanders, Belgium. Chemosphere, 81, 478-487. Davis, J. W., S. J. Gonsoir, D. A. Markham, U. R. S. Friederich, R. W. Hunziker, and J. M. Ariano. 2006. Biodegradation and Product Identification of [14c]Hexabromocyclododecane in Wastewater Sludge and Freshwater Sediment. Environmental Science and Technology, 40, 5395-5401. De Boer, J., C. Allchin, B. Zegers, J. P. Boon, S. H. Brandsma, S. Morris, A. W. Kruij't, J. M. van der Hesselingen, and J. J. Haftka. 2002. HBCD and TBBP-A in Sewage Slugde, Sediments and Biota, Including Interlaboratory Study; Co33/02. RIVO-Netherlands Institute for Fisheries Researach, Wageningen, The Netherlands. de Boer, J., H. A. Leslie, P. E. G. Leonards, P. Bersuder, S. Morris, and C. R. Allchin (Abstract. The Third International Workshop on Brominated Flame Retardants, June 6-9, 2004). 2004. Screening and Time Trend Study of Decabromodiphenylether and Hexabromocyclododecane in Birds. Toronto, Canada, (as cited in UNEP, 2010). de Winter-Sorkina, R., M. I. Bakker, G. Wolterink, and M. J. Zeilmaker. 2006. Brominated Flame Retardants: Occurrence, Dietary Intake and Risk Assessment. RIVM-report 320100002/2006. Centre for Substances and Integrated Risk Assessment, National Institute for Public Health and the Environment (RIVM), The Netherlands. http://rivm.openrepositorv.eom/rivm/bitstream/10029/7303/l/320100002.pdf. de Wit, C. A., J. A. Bjorklund, and K. Thuresson. 2012. Tri-Decabrominated Diphenyl Ethers and Hexabromocyclododecane in Indoor Air and Dust from Stockholm Microenvironments 2: Indoor Sources and Human Exposure. Environment International, 39,141-147. Page 41 of 97 ------- Deng, J., L Yu, C. Liu, K. Yu, X. Shi, L W. Y. Yeung, R. S. S. Lam, and B. Zhou. 2009. Hexabromocyclododecane-lnduced Developmental Toxicity and Apoptosis in Zebrafish Embryos. Aquatic Toxicology, 93(1), 29-36. DEP (Maine Department of Environmental Protection). 2013. Chemicals of High Concern. Augusta, Maine, http://www.maine.gov/dep/safechem/highconcern/ (accessed on January 23, 2015). DEQ (Oregon Department of Environmental Quality). 2010a. Priority Persistent Pollutant List (P3 List). Portland, OR. http://www.deq.state.or.us/wq/SB737/docs/LegRpAtt20100601.pdf (accessed on January 23, 2015). DEQ (Oregon Department of Environmental Quality). 2010b. Pollutant Profiles. Portland, Oregon. http://www.deq.state.or.us/wq/SB737/docs/LegRpAtt420100601.pdf (accessed on January 23, 2015). DEQ (Oregon Department of Environmental Quality). 2011. Water Quality Senate Bill 737. http://www.deq.state.or.us/wq/SB737/ (accessed on January 23, 2015). Desjardins, D., J. MacGregor, and H. Krueger. 2004. Final Report: Hexabromocyclododecane (HBCD): A 72-Hour Toxicity Test with the Marine Diatom (Skeletomema Costatum). Project Number: 439A-125. Wildlife International Ltd for Chemical Manufacturers Association, Easton, MD. (As cited in EPA (2014) Flame retardant alternatives for Hexabromocyclododecane (HBCD)). Desjardins, D., J. MacGregor, and H. Krueger. 2005. Final Report. Hexabromocyclododencane (HBCD): A 72-Hour Toxicity Test with the Marine Diatom (Skeletonema Costatum) Using a Co-Solvent, Chapter 2. Wildlife International Ltd for Chemical Manufacturers Association, Easton, MD. (As cited in EPA (2014) Flame retardant alternatives for Hexabromocyclododecane (HBCD)). Devanathan, G., A. Subramanian, A. Sudaryanto, S. Takahashi, T. Isobe, and S. Tanabe. 2012. Brominated Flame Retardants and Polychlorinated Biphenyls in Human Breast Milk from Several Locations in India: Potential Contaminant Sources in a Municipal Dumping Site. Environment International, 39, 87-95. Dodson, R. E., L. J. Perovich, A. Covaci, N. VandenEade, A. C. lonas, A. C. Dirtu, J. G. Brody, and R. A. Rudel. 2012. After the PBDE Phase-Out: A Broad Suite of Flame Retardants in Repeat House Dust Samples from California. Environmental Science and Technology, 46(24), 13056-13066. Drottar, K. R., and H. O. Krueger. 1998. Hexabromocyclododecane (HBCD): A Flow-through Life- Cycle Toxicity Test with the Cladoceran (Daphnia Magna). Final Report. OTS0559490. Wildlife International Ltd for Chemical Manufacturers Association, Easton, MD. Page 42 of 97 ------- Drottar, K. R., and H. O. Krueger. 2000. Hexabromocyclododecane (HBCD): A Flow-through Bioconcentration Test with the Rainbow Trout (Oncorhynchus Mykiss). Final Report. 2000. OTS0559424. Wildlife International Ltd for Chemical Manufacturers Association, Easton, MD. DISC (State of California Department of Toxic Substances Control). 2010. Scp Candidate Chemical Hexabromocyclododecane [25637-99-4]. Safer Consumer Products, Sacramento, CA.. https://dtsc.ca.gov/SCP/ChemList.cfm (accessed on January 23, 2015). EC (European Commission, European Chemicals Bureau). 2008. Risk Assessment: Hexabromocyclododecane. Office for Official Publications of the European Communities, Luxembourg. http://ec.europa.eu/health/ph risk/committees/04 scher/docs/scher o 093.pdf. ECHA (European Chemicals Agency). 2009. Data on Manufacture, Import, Export, Uses and Releases of HBCDD as Well as Information on Potential Alternatives to Its Use. ECHA/2008/2. IOM Consulting, supported by BRE, PFAand Entec., Helsinki, Finland. http://echa.europa.eu/doc/consultations/recommendations/tech reports/tech rep hb cdd.pdf. Eguchi, A., T. Isobe, K. Ramu, N. M. Tue, A. Sudaryanto, G. Devanathan, P. H. Viet, R. S. Tana, S. Takahashi, A. Subramanian, and S. Tanabe. 2013. So/7 Contamination by Brominated Flame Retardants in Open Waste Dumping Sites in Asian Developing Countries. Chemosphere, 90, 2365-2371. Ema, M., S. Fujii, M. Hirata-Koizumi, and M. Matsumoto. 2008. Two-Generation Reproductive Toxicity Study of the Flame Retardant Hexabromocyclododecane in Rats. Reproductive Toxicology, 25, 335-351. Environment CA and Health CA (Environment Canada and Health Canada). 2011. Screening Assessment Report on Hexabromocyclododecane . Chemical Abstracts Service Registry Number 3194-55-6., Ottawa, Canda. http://www.ec.gc.ca/ese- ees/default.asp?lang=En&n=7882C148-l#a4. EPA (US Environmental Protection Agency). 1991. Guidelines for Developmental Toxicity Risk Assessment. EPA/600/FR-91/001. Risk Assessment Forum, Washington, DC. http://www.epa.gov/iris/backgrd.htm (accessed January 16, 2009). EPA (US Environmental Protection Agency). 1993. Determination of Rates of Reaction in the Gas-Phase in the Troposphere. 5. Rate of Indirect Photoreaction: Evaluation of the Atmospheric Oxidation Computer Program of Syracuse Research Corporation for Estimating the Second-Order Rate Constant for the Reaction of an Organic Chemical with Page 43 of 97 ------- Hydroxyl Radicals. EPA-744-R-93-001. Office of Pollution Prevention and Toxics, Washington, DC. EPA (US Environmental Protection Agency). 1996. Guidelines for Reproductive Toxicity Risk Assessment. EPA/630/R-96/009. Risk Assessment Forum, Washington, DC. http://www.epa.gov/iris/backgrd.htm (accessed January 16, 2009). EPA (US Environmental Protection Agency). 1998a. Guidelines for Ecological Risk Assessment. EPA/630/R-95/002F. Office of the Science Advisor, Risk Assessment Forum, Washington, DC. http://www2.epa.gov/sites/production/files/2014- 11/documents/eco risk assessmentl998.pdf. EPA (US Environmental Protection Agency). 1998b. Guidelines for Neurotoxicity Risk Assessment. EPA/630/R-95/001F. Risk Assessment Forum, Washington, DC. http://www.epa.gov/iris/backgrd.html (accessed January 16, 2009). EPA (US Environmental Protection Agency). 2006a. Non-Confidential 2006 Inventory Update Reporting (IUR) Database. Office of Pollution Prevention and Toxics, Washington, DC. http://cfpub.epa.gov/iursearch/. EPA (US Environmental Protection Agency). 2007. Exposure and Fate Assessment Screening Tool (E-Fast). Version 2.0. Documentation Manual. Office of Pollution Prevention and Toxics, Washington, DC. www.epa.gov/oppt/exposure/pubs/efast2man.pdf. EPA (US Environmental Protection Agency). 2008a. Initial Risk-Based Prioritization of High Production Volume Chemicals. Chemical/Category: Hexabromocyclododecane (HBCD). Office of Pollution Prevention and Toxics, Washington, DC. http://www.epa.gov/hpvis/rbp/HBCD.3194556.Web.RBP.31308.pdf. EPA (US Environmental Protection Agency). 2010a. Hexabromocyclododecane (HBCD) Action Plan. Office of Pollution Prevention and Toxics, Washington, DC. http://www.epa.gov/oppt/existingchemicals/pubs/actionplans/RIN2070- AZ10 HBCD%20action%20plan Final 2010-08-09.pdf. EPA (US Environmental Protection Agency). 2011a. Epi Suite Results for CASRN 3194-55-6. Download Epi Suite Tm V4.0. Office of Pollution Prevention and Toxics, Washington, DC. http://www.epa.gov/opptintr/exposure/pubs/episuitedl.htm. EPA (US Environmental Protection Agency). 2011b. Exposure Factors Handbook: 2011 Edition. National Center for Environmental Assessment, Washington, DC. http://www.epa.gov/ncea/efh/pdfs/efh-complete.pdf. Page 44 of 97 ------- EPA (US Environmental Protection Agency). 2012a. Non-Confidential 2012 Chemical Data Reporting (CDR) Database. Office of Pollution, Prevention and Toxics, Washington, DC. http://www.epa.gov/cdr/. EPA (US Environmental Protection Agency). 2012e. Significant New Use Rule for Hexabromocyclododecane and 1,2,5,6,9,10-Hexabromocyclododecane. 77 Federal Register 58 (March 26, 2012), pp. 17386-17394. EPA (US Environmental Protection Agency). 2014a. Estimated Fish Consumption Rates for the U.S. Population and Selected Subpopulations (Nhanes 2003-2010). . EPA-820-R-14-002. Final Report., Washington, DC. EPA (US Environmental Protection Agency). 2014b. Flame Retardant Alternatives to Hexabromocyclododecane (HBCD). EPA/740R14001. Office of Pollution Prevention and Toxics, Design for the Environment, Washington, DC. http://www.epa.gov/dfe/pubs/proiects/hbcd/hbcd-full-report-508.pdf. EPA (US Environmental Protection Agency). 2014c. Framework for Human Health Risk Assessment to Inform Decision Making. EPA/100/R-14/001. Office of the Science Advisor, Washington, DC. http://www2.epa.gov/sites/production/files/2014- 12/documents/hhra-f ramework-final-2014.pdf. EPA (US Envrionmental Protection Agency). 2014d. Preliminary Materials for the Integrated Risk Information System (IRIS) Toxicological Review of Hexabromocyclododecane (HBCD) EPA/630/R-13/235. National Center for Environmental Assessment, Office of Research and Development, Washington, DC. http://www.epa.gov/iris/publicmeeting/iris bimonthly-apr2014/HBCD- preliminary draft materials.pdf. (as cited in IRIS Program Progress Report and Report to Congress, February, 2015). EPA (US Environmental Protection Agency). 2015a. Drinking Water Contaminant Candidate List (CCL) and Regulatory Determination. Office of Water, Washington, DC. http://www2.epa.gov/ccl (accessed on May 12, 2015). EPA (US Environmental Protection Agency). 2015b. Drinking Water Contaminants. Office of Water, Washington, DC. http://water.epa.gov/drink/contaminants/ (accessed on May 12, 2015). EPA (US Environmental Protection Agency). 2015c. Iristrack Detailed Report. Integrated Risk Information System, Washington, DC. http://cfpub.epa.gov/ncea/iristrac/index.cfm?fuseaction=viewChemical.showChemical& sw id=1102 (accessed on March 5, 2015). Page 45 of 97 ------- Eriksson, P., C. Fischer, M. Wallin, E. Jakobsson, and A. Frederiksson. 2006. Impaired Behaviour, Learning and Memory, in Adult Mice Neonatally Exposed to Hexabromocyclododecane (HBCDD). Environmental Toxicology and Pharmacology, 21(3), 317-322. Ethyl Corporation. 1985a. Genetic Toxicology Salmonella/Microsomal Assay on HBCD Cover letter dated 030890. Submitted under TSCA Section 8D; EPA Document No. 86- 900000164; NTIS No. OTS0522235. Washington, DC. Ethyl Corporation. 1985b. Genetic Toxicology Rat Hepatocyte Primary Culture/DNA Repair Test on HBCD Cover letter dated 030890. Submitted under TSCA Section 8D; EPA Document No. 86-900000163; NTIS No. OTS0522234. Washington, DC. Extruded Polystyrene Foam Association (Polystyrene Foam Association). Extruded Polystyrene Foam (XPS) Thermal Insulation Overview. Presentation: EPA (US Environmental Protection Agency), 2011. Presentation on XPS as cited in Flame Retardant Alternatives for Hexabromocyclododecane (HBCD), US EPA, 2014. Fangstrom, B., I. Athanassiadis, T. Odsjo, and et al. 2008. Temporal Trends of Polybrominated Diphenyl Ethers and HBCD in Milk from Stockholm Mothers, 1980-2004. Mol Nutr Food Res, 52(2), 187-193. Fangstrom, B., A. Strid, and A. Bergman. 2005. Temporal Trends ofBrominated Flame Retardants in Milk from Stockholm Mothers, 1980-2004. Department of Environmental Chemistry, Stockholm University, Stockholm, Sweden. http://www.imm.ki.se/Datavard/PDF/mi%C3%B6lk poolade NV%20rapport%202005% 20modersmiolk.pdf (accessed November 21, 2008). Feng, A.-H., S.-J. Chen, M.-Y. Chen, M.-J. He, X.-J. Luo, and B.-X. Mai. 2012. Hexabromocyclododecane (HBCD) and Tetrabromobisphenol a (TBBPA) in Riverine and Estuarine Sediments of the Pearl River Delta in Southern China, with Emphasis on Spatial Variability in Diastereoisomer- and Enantiomer-Specific Distribution of HBCD. Marine Pollution Bulletin, 64, 919-925. Friddle, J. (Eagle Performance Products). Use of HBCD in Textiles. Personal communication.: Lavoie., E. (US Environmental Protection Agency), February 8. Discussion of the use of HBCD in textiles. Gerecke, A. C., W. Giger, P. C. Hartmann, N. V. Heeb, H. P. Kohler, P. Schmid, M. Zennegg, and M. Kohler. 2006. Anaerobic Degradation ofBrominated Flame Retardants in Sewage Sludge. Chemosphere, 64(2), 311-317. Gheorghe, A., A. C. Dirtu, H. Neels, and A. Covaci. 2013. Brominated and Organophosphate Flame Retardants in Indoor Dust from Southern Romania. San Francisco, CA. www.bfr2013.com. Page 46 of 97 ------- Gorga, M., E. Martinez, A. Ginebreda, E. Eljarrat, and D. Barcelo. 2013. Determination of PBDEs, Hbb, Pbeb, Dbdpe, HBCD, TBBPA and Related Compounds in Sewage Sludge from Catalonia (Spain). Science of the Total Environment, 444, 51-59. Graves, W. C, and J. P. Swigert. 1997a. Hexabromocyclododecane (HBCD): A 48-Hour Flow- through Acute Toxicity Test with the Cladoceran (Daphnia Magna). Report# 439A-102. TSCA Section 8D; OTS0559002 , DCN 86970000792. Wildlife International Ltd, Easton, MD. Graves, W. C., and J. P. Swigert. 1997b. Hexabromocyclododecane (HBCD): A 96-Hour Flow- through Acute Toxicity Test with the Rainbow Trout (Oncorhynchus Mykiss). Report# 439A-101, pp 70 pp. TSCA Section 8D; OTS0559424, DCN 86980000086. Wildlife International Ltd, Easton, MD. Guerra, P., M. Alaee, B. Jimenez, G. Pacepavicius, C. Marvin, G. Maclnnis, E. Eljarrat, L. Champoux, and K. Fernie. 2012. Emerging and Historical Brominated Flame Retardants in Peregrine Falcon (Falco Peregrinus) Eggs from Canada and Spain. Environment International, 40,179-186. Guerra, P., E. Eljarrat, and D. Barcelo. 2010. Simultaneous Determination of Hexabromocyclododecane, Tetrabromobisphenol a, and Related Compounds in Sewage Sludge and Sediment Samples from Ebro River Basin (Spain). Analytical and Bioanalytical Chemistry, 397, 2817-2824. Gulf South Research Institute. 1988. Initial Submission: Letter from Ethyl Corp to USEPA Re Technical and Toxicity Data on Brominated Flame Retardants Including HBCD. EPA Document No. FYI-OTS-0794-0947; NTIS No. OTS0000947. Washington, DC. Hakk, H., D. T. Szabo, J. Huwe, J. Diliberto, and L. S. Birnbaum. 2012. Novel and Distinct Metabolites Identified Following a Single Oral Dose of A- or F-Hexabromocyclododecane in Mice. Environmental Science &Technology, 46(1), 13494-13503. Harrad, S., and M. A.-E. Abdallah. 2011. Brominated Flame Retardants in Dust from UK Cars - within-Vehicle Spatial Variability, Evidence for Degradation and Exposure Implications. Chemosphere, 82(9), 1240-1245. Harrad, S., M. A.-E. Abdallah, N. L. Rose, S. D. Turner, and T. A. Davidson. 2009. Current-Use Brominated Flame Retardants in Water, Sediment, and Fish from English Lakes. Environmental Toxicology and Chemistry, 43(24), 9077-9083. Harrad, S., E. Goosey, J. Desborough, M. A. E. Abdallah, L. Roosens, and A. Covaci. 2010. Dust from U.K. Primary School Classrooms and Daycare Centers: The Significance of Dust as a Page 47 of 97 ------- Pathway of Exposure of Young U.K. Children to Brominated Flame Retardants and Polychlorinated Biphenyls. Environmental Science & Technology, 44, 4198-4202. Harscher, M. (Unknown). Uses of Hexabromocyclododecane (HBCD); Flammability Standards. Meeting: EPA (US Environmental Protection Agency, Crystal City, VA), 2011. As cited in Flame Retardant Alternatives for Hexabromocyclododecane (HBCD), US EPA, 2014. He, M.-J., X.-J. Luo, L.-H. Yu, J.-P. Wu, S.-J. Chen, and B.-X. Mai. 2013. Diasteroisomer and Enantiomer-Specific Profiles of Hexabromocyclododecane and Tetrabromobisphenol a in an Aquatic Environment in a Highly Industrialized Area, South China: Vertical Profile, Phase Partition, and Bioaccumulation. Environmental Pollution, 179. He, S., M. Li, J. Jin, Y. Wang, Y. Bu, M. Xu, X. Yang, and A. Liu. 2013. Concentrations and Trends of Halogenated Flame Retardants in the Pooled Serum of Residents of Laizhou Bay, China. . Environmental Toxicology and Chemistry, 32(6), 1242-1247. Helleday, T., K. L. Tuominen, A. Bergman, and et al. 1990. Brominated Flame Retardants Induce Intragenic Recombination in Mammalian Cells. Mutation Research, 439(2), 137-147. Huntington Research Center. 1978. Ames Metabolic Activation Test to Assess the Potential Mutagenic Effect of Compound No. 49. Cover letter dated 031290. Submitted under TSCA Section 8D; EPA Document No. 86-900000385; NTIS No. OTS0522948. Washington, DC. Hwang, I.-K., H.-H. Kang, I.-S. Lee, and J.-E. Oh. 2012. Assessment of Characteristic Distribution of Pcdd/Fs and BFRs in Sludge Generated at Municipal and Industrial Wastewater Treatment Plants. Chemosphere, 88, 888-894. IBT (Industrial Bio-Test Labs). 1990. Mutagenicity of Two Lots of Fm-100 Lot 53 and Residue of Lot 3322 in the Absence and Presence of Metabolic Activation Test data and cover letter. Submitted under TSCA Section 8D; EPA Document No. 86-900000267; NTIS No. OTS0523259. Washington, DC. IRDC (International Research and Development Corporation). 1977. Acute Toxicity Studies in Rabbits and Rats with HBCD. Submitted under TSCA Section 8E; EPA Document No. 88- 7800065; NTIS No. OTS0200051. Washington, DC. IRDC (International Research and Development Corporation). 1978a. Acute Toxicity Studies in Rabbits and Rats with HBCD. Attachments and cover letter dated 030178. Submitted under TSCA Section 8E; EPA Document No. 88-7800088; NTIS No. OTS0200466. Washington, DC. IRDC (International Research and Development Corporation). 1978b. Acute Inhalation Toxicity Study in Rats with HBCD. Attachments and cover letter dated 042478. Submitted under Page 48 of 97 ------- TSCA Section 8E; EPA Document No. 88-7800137; NTIS No. OTS0200488. Washington, DC. Ismail, N., S. B. Gewurtz, K. Pleskach, D. M. Whittle, P. A. Helm, C. H. Marvin, and G. T. Tomy. 2009. Brominated and Chlorinated Flame Retardants in Lake Ontario, Canada, Lake Trout (Salvelinus Namaycush) between 1979 and 2004 and Possible Influences of Food- Web Changes. Environmental Toxicology and Chemistry, 28(5), 910-920. Jatzek. 1990. Determination of the Acute Toxicity of Hexabromid S to the Waterflea Daphnia Magna Straus. 1184/88, BASF Indolfab. TSCA Section 8D; OTS0522955, DCN 86900000392. BASF. Johnson-Restrepo, B., D. H. Adams, and K. Kannan. 2008. Tetrabromobisphenol A (TBBPA) and Hexabromocyclododecanes (HBCDs) in Tissues of Humans, Dolphins, and Sharks from the United States. Chemosphere, 70(1), 1935-1944. Johnson, P. I., H. M. Stapleton, B. Mukherjee, R. Mauser, and J. D. Meeker. 2013. Associations between Brominated Flame Retardants in House Dust and Hormone Levels in Men. Science of the Total Environment, 445,177-184. Kajiwara, N., and H. Takigami. 2013. Behavior of Additive Brominated Flame Retardants in Textile Products. 5th International Symposium on Brominated Flame Retardants, April 07-April 09, 2010, Kyoto, Japan, http://www.bfr2010.com/. Kakimoto, K., K. Akutsu, Y. Konishi, and et al. 2008. Time Trend ofHBCD in the Breast Milk of Japanese Women. Chemosphere, 71(6), 1110-1114. Kalachova, K., P. Hradkova, D. Lankova, J. Hajslova, and J. Pulkrabova. 2012. Occurrence of Brominated Flame Retardants in Household and Car Dust from the Czech Republic. Science of the Total Environment, 441,182-193. Kicinski, M., M. K. Viaene, E. Den Hond, G. Schoeters, A. Covaci, A. C. Dirtu, V. Nelen, L Bruckers, K. Croes, I. Sioen, W. Baeyens, N. Van Larebeke, and T. S. Nawrot. 2012. Neurobehavioral Function and Low-Level Exposure to Brominated Flame Retardants in Adolescents: A Cross-Sectional Study. Environmental Health, 11(1), 86. Klosterhaus, S. L, H. M. Stapleton, M. J. La Guardia, and D. J. Greig. 2012. Brominated and Chlorinated Flame Retardants in San Francisco Bay Sediments and Wildlife. . Environment International, 47, 56-65. Kobiliris, D. 2010. Influence of Embryonic Exposure to Hexabromocyclododecane (HBCD) on the Corticosterone Response and "Fight or Flight" Behaviors If Captive American Kestrels. McGill University, Montreal, Canada. Accessed at http://www.researchgate.net. Page 49 of 97 ------- Kopp, E. K., H. Fromme, and W. Voelkel. 2012. Analysis of Common and Emerging Brominated Flame Retardants in House Dust Using Ultrasonic Assisted Solvent Extraction and on-Line Sample Preparation Via Column Switching with Liquid Chromatography-Mass Spectrometry. Journal of Chromatography A, 1241, 28-36. Kuiper, R. V., R. F. Canton, P. E. Leonards, B. M. Jenssen, M. Dubbeldam, P. W. Wester, d. B. M. van, J. G. Vos, and A. D. Vethaak. 2007. Long-Term Exposure of European Flounder (Platichthys Flesus) to the Flame-Retardants Tetrabromobisphenol a (TBBPA) and Hexabromocyclododecane (HBCD). Ecotoxicology and Environmental Safety, 67(3), 349- 360. Kuo, Y.-Y., H. Zhang, A. C. Gerecke, and J. Wang. 2014. Chemical Composition of Nanoparticles Released from Thermal Cutting of Polystyrene Foams and the Associated Isomerization of Hexabromocyclododecane (HBCD) Diastereomers. Aerosol and Air Quality Research, 14(4), 1114-1120. La Guardia, M., R. Hale, E. Harvey, T. Mainor, and S. Ciparis. 2012. In Situ Accumulation of HBCD, PBDEs and Several Alternative Flame Retardants in the Bivalve (Corbicula Fluminea) and Gastropod (Elimia Proximo). Environmental Science and Technology, 46(11), 5798-5805. La Guardia, M. J., R. C. Hale, E. Harvey, and D. Chen. 2010. Flame-Retardants and Other Organohalogens Detected in Sewage Sludge by Electron Capture Negative Ion Mass Spectrometry. Environmental Science & Technology, 44(1), 4658-4664. La Guardia, M. J., R. C. Hale, and B. Newman. 2013. Brominated Flame-Retardants in Sub- Saharan Africa: Burdens in Inland and Coastal Sediments of the Ethekwini Metropolitan Municipality, South Africa. Environmental Science & Technology, 47, 9643-9650. Law, K., T. Halldorson, R. Danell, G. Stern, S. B. Gewurtz, M. Alaee, C. Marvin, M. Whittle, and G. To my. 2006a. Bioaccumulation and Trophic Transfer of Some Brominated Flame Retardants in a Lake Winnepeg (Canada) Food Web. Environmental Toxicology and Chemistry, 25(8), 2177-2186. Law, K., V. P. Palace, T. Halldorson, R. Danell, K. Wautier, B. Evans, M. Alaee, C. Marvin, and G. T. Tomy. 2006b. Dietary Accumulation of Hexabromocyclododecane Diastereoisomers in Juvenile Rainbow Trout (Oncorhynchus Mykiss) I: Bioaccumulation Parameters and Evidence ofBioisomerization. Environmental Toxicology and Chemistry, 25(7), 1757- 1761. Law, R. J., A. Covaci, S. Harrad, D. Herzke, M. A.-E. Abdallah, K. Fernie, L M. Toms, and H. Takigami. 2014. Levels and Trends of PBDEs and HBCDs in the Global Environment: Status at the End of 2012. Environment International, 65,147-158. Page 50 of 97 ------- Lewis, A. C, and A. L. Palanker. 1978. A Dermal Ld50 Study in Albino Rabbits and an Inhalation Lc50 Study in Albino Rats. Experiment Reference No. 78385-2. Client: Saytech Inc.. Consumer Product Testing, Fairfield, NJ. Li, H., L. Mo, Z. Yu, G. Sheng, and J. Fu. 2012a. Levels, Isomer Profiles and Chiral Signatures of Particle-Bound Hexabromocyclododecanes in Ambient Air around Shanghai, China. Environmental Pollution, 165,140-146. Li, H., H. Shang, P. Wang, Y. Wang, H. Zhang, Q. Zhang, and G. Jiang. 2013. Occurrence and Distribution of Hexabromocyclododecane in Sediments from Seven Major River Drainage Basins in China. Journal of Environmental Sciences, 25(1), 69-76. Li, H., Q. Zhang, P. Wang, Y. F. Li, J. Lv, W. Chen, D. Geng, Y. Wnag, T. Wang, and G. Jiang. 2012b. Levels and Distribution of Hexabromocyclododecane (HBCD) in Environmental Samples near Manufacturing Facilities in Laizhou Bay Area, East China. Journal of Environmental Monitoring, 14, 2591-2597. Lilienthal, H., d. V. L. T. van, A. H. Piersma, and J. G. Vos. 2009. Effects of the Brominated Flame Retardant Hexabromocyclododecane (HBCD) on Dopamine-Dependent Behavior and Brainstem Auditory Evoked Potentials in a One-Generation Reproduction Study in Wistar Rats. Toxicology Letters, 185, 63-72. Lindberg, P., U. Sellstrom, L. Haggberg, and C. A. deWitt. 2004. Higher Brominated Diphenyl Ethers and Hexabromocyclododecane Found in Eggs of Peregrine Falcons (Falco Peregrines) Breeding in Sweden. Environmental Science & Technology, 38(1), 93-96. Litton Bionetics Inc. 1990. Mutagenicity Evaluation of 421-32b (Final Report) With test data and cover letter. Submitted under TSCA Section 8D; EPA Document No. 86-900000265; NTIS No. OTS0523257. Washington, DC. Malarvannan, G., T. Isobe, A. Covaci, M. Prudente, and S. Tanabe. 2013. Accumulation of Brominated Flame Retardants and Polychlorinated Biphenyls in Human Breast Milk and Scalp Hair from the Philippines: Levels, Distribution and Profiles. Science of the Total Environment, 442, 366-379. Managaki, S., I. Enomoto, and S. Masunaga. 2012. Sources and Distribution of Hexabromocyclododecanes (HBCDs) in Japanese River Sediment. Journal of Environmental Monitoring, 14, 901-907. Marvin, C. H., G. T. Tommy, J. A. Armitage, J. A. Arnot, L. McCarthy, A. Covaci, and V. Palace. 2011. Hexabromocyclododecane: Current Understanding of Chemistry, Environmental Fate and Toxicology and Implications for Global Management. Environmental Science and Technology, 45, 8613-8623. Page 51 of 97 ------- Maul, J., B. G. Frushour, J. R. Kontoff, H. Eichenauer, K.-H. Ott, and C. Schade. 2007. Polystyrene and Styrene Copolymers. In Bellussi, G. e. a., Ullmann's Encyclopedia of Industrial Chemistry (Vol. 29, pp. 475-522). Wiley-VCH Verlag GmbH & Co. KGaA, Aachen, Germany, http://dx.doi.org/10.1002/14356007.a21 615.pub2. MDH (Minnesota Department of Health). 2013. Chemicals of High Concern List. St Paul, MN. http://www.health.state.mn.us/divs/eh/hazardous/topics/toxfreekids/chclist/mdhchc2 013.pdf (accessed on January 23, 2015). Meijer, L, M. Van Velzen, and et al. 2008. Serum Concentrations of Neutral and Phenolic Organohalogens in Pregnant Women and Some of Their Infants in the Netherlands. Environmental Science &Technology, 42(9), 3428-3433. Microbiological Associates Inc. 1996. HBCD (HBCD): Chromosome Aberrations in Human Peripheral Blood Lymphocytes with Cover Letter Dated 12/12/1996. Submitted under TSCA Section 8D; EPA Document No. 86970000358; NTIS No. OTS0573552. Washington, DC. MOE (Ministry of Environment, Japan). 2000. Chemicals in the Environment, Report on Environmental Survey and Wildlife Monitoring of Chemicals in F. Y. 1998. Environmental Health and Safety Division, Tokyo, Japan. MOE (Ministry of the Environment, Government of Japan). 2005. Chemicals in the Environment Report on Environmental Survey and Monitoring of Chemicals in Fy2004. Environment Health and Safety Division, Environment Health Department Tokyo, JAPAN. MOEJ (Ministry of the Environment, Japan). 2009. 6-Week Administration Study of 1,2,5,6,9,10- Hexabromocyclododecane for Avian Reproduction Toxicity under Long-Day Conditions Using Japanese Quail. Research Institute for Animal Science in Biochemistry & Toxicology Tokyo, Japan. http://chm.pops.int/Convention/POPsReviewCommittee/hrPOPRCMeetings/POPRC5/P OPRC5Followupcommunications/HBCDInvitationforcommentsondraftRP/tabid/742/lang uage/en-US/Default.aspx. (as cited in Hexabromocyclododecane Draft Risk Profile in relation to the Stockholm Convention on Persistent Organic Pollutants). Momma, J., M. Kaniwa, H. Sekiguchi, K. Ohno, Y. Kawasaki, M. Tsuda, A. Nakamura, and Y. Kurokawa. 1993. Dermatological Evaluation of a Flame Retardant, Hexabromocyclododecane (HBCD) on Guinea Pig by Using the Primary Irritation, Sensitization, Phototoxicity and Photosensitization of Skin. Eisei Shikenjo Hokoku, 111(1), 18-24. Morose, G. (Prepared for the Jennifer Altman Foundation.). 2006. An Overview of Alternatives to Tetrabomobisphenol a (TBBPA) and Hexabromocyclododecane (HBCD). Lowell Center Page 52 of 97 ------- for Sustainable Production, University of Massachusetts, Lowell., Lowell, MA. http://www.chemicalspolicv.org/downloads/AternativestoTBBPAandHBCD.pdf. Morris, S., C. R. Allchin, B. N. Zegers, J. J. H. Haftka, J. P. Boon, C. Belpaire, P. E. G. Leonards, S. P. J. Van Leeuwen, and J. De Boer. 2004. Distribution and Fate ofHBCD and TBBPA Brominated Flame Retardants in North Sea Estuaries and Aquatic Food Webs. Environmental Science &Technology, 38(21), 5497-5504. Muir, D. C. G., S. Backus, A. E. Derocher, R. Dietz, T. J. Evans, G. W. Gabrielsen, J. Nagy, R. J. Norstrom, C. Sonne, I. Stirling, M. K. Taylor, and R. J. Letcher. 2006. Brominated Flame Retardants in Polar Bears (Ursus Maritimus)from Alaska, the Canadian Arctic, East Greenland, andSvalbard. Environmental Science &Technology, 40(2), 449-455. (as cited in UNEP, 2010). Murai, T., H. Kawasaki, and S. Kanoh. 1985. Studies on the Toxicity of Insecticides and Food Additives in Pregnant Rats-Fetal Toxicity of Hexabromocyclododecane. Pharmacometrics, 29(6), 981-986. Ni, H.-G., and H. Zeng. 2013. HBCD and TBBPA in Particulate Phase of Indoor Air in Shenzhen, China. Science of the Total Environment, 458-460,15-19. NICNAS (National Industrial Chemicals Notification and Assessment Scheme). 2012. Hexabromocyclododecane. PEC34. Priority existing chemical assessment report. Volume 34, Government of Australia., Canberra, Australia. http://www.nicnas.gov.au/Publications/CAR/PEC/PEC34/HBCD Report June 2012 PDF .pdf. OECD (Organisation for Economic Co-operation and Development). 1983. Guideline for the Testing of Chemicals: One-Generation Reproduction Toxicity Study, Guideline 415. Paris, France, http://www.oecd.org/chemicalsafety/risk-assessment/1948458.pdf (accessed January 14, 2014). OECD (Organisation for Economic Co-operation and Development). 1995. Guideline for the Testing of Chemicals: Repeated Dose 28-Day Oral Toxicity Study in Rodents, Guideline 407. Paris, France, http://www.oecd.org/env/ehs/testing/37477972.pdf (accessed January 14, 2014). OECD (Organisation for Economic Co-operation and Development). 2001. Guideline for the Testing of Chemicals: Two-Generation Reproduction Toxicity Study, Guideline 416. Paris, France, http://www.oecd.org/chemicalsafety/risk-assessment/1948466.pdf (accessed January 14, 2014). Page 53 of 97 ------- OECD (Organisation for Economic Co-operation and Development). 2004b. Emission Scenario Document on Plastics Additives. OECD Series On Emission Scenario Documents,, Paris, France. OECD (Organisation for Economic Co-operation and Development). 2007. SIDS Intial Assessment Profile for HBCDD. OECD Existing Chemicals Database, Paris, France. http://webnet.oecd.ore/HPV/UI/handler.axd?id=ea58acll-e090-4b24-b281- 200ae351686c. OEHHA (State of California Environmental Protection Agency, Office of Environmental Health Hazard Assessment). 2007. Proposition 65 in Plain Language. Sacramento, California. http://oehha.ca.gov/prop65/background/p65plain.html (accessed on January 23, 2015). OEHHA (Office of Environmental Health Hazard Assessment). 2014. Chemicals Known to the State to Cause Cancer or Reproductive Toxicity, May 2, 2014. State of California Environmental Protection Agency,, Sacramento, California. http://oehha.ca.gov/prop65/prop65 list/files/P65single050214.pdf (accessed on January 23, 2015). Oetken, M., K. Ludwichowski, and R. Nagel. 2001. Validation of the Preliminary EU-Concept of Assessing the Impact of Chemicals to Organisms in Sediment by Using Selected Substances. Institute of Hydrobiology UBA-FB 299 67 411, pp 97. Dresden University of Technology, Dresden, Germany, (as cited in EC, 2008). Owens Corning. 2005. Material Safety Data Sheet Foamular Extruded Polystyrene Insulation. 15- MSD-21528-01-D. Product Stewardship, Toledo, OH. http://www.actiocms.com/VIEW MSDS/AuthorDisplay V402/msdsdisplaycode author new MASTER.cfm?edit msds id=19186&dbname=production&Hide Section Number s=N&formatcode=14&language=l&noprint label fax email=Y. Palace, V., B. Park, K. Pleskach, B. Gemmill, and G. Tomy. 2010. Altered Thyroxine Metabolism in Rainbow Trout (Oncorhynchus My kiss) Exposed to Hexabromocyclododecane (HBCD). Chemosphere, 80(2), 165-169. Palace, V. P., K. Pleskach, T. Halidorson, R. Danell, K. Watier, B. Evans, M. Alaee, C. Marvin, and G. T. Tommy. 2008. Biotransformation Enzymes and Thyroid Axis Disruption in Juvenile Rainbow Trout (Oncorhynchus Mykiss) Exposed to Hexabromocyclododecane Diastereoisomers. Environmental Science and Technology, 42,1967-1972. Peck, A. M., R. S. Pugh, A. Moors, M. B. Ellison, B. J. Porter, P. R. Becker, and J. R. Kucklick. 2008. Hexabromocyclododecane in White-Sided Dolphins: Temporal Trend and Stereoisomer Distribution in Tissues. Environmental Science &Technology, 42(7), 2650-2655. Page 54 of 97 ------- Pharmakoligisches Institute. 1978. Ames Test with Hexabromides with Cover Letter Dated 031290. Submitted under TSCA Section 8D; EPA Document No. 86-900000379; NTIS No. OTS0522942. Washington, DC. Porch, J. R., T. Z. Kendall, and H. O. Krueger. 2002. Hexabromocyclododecane (HBCD): A Toxicity Test to Determine the Effects of the Test Substance on Seedling Emergence of Six Species of Plants. 126 pp. Wildlife International Ltd, Easton, MD. (as cited in EC, 2008). Pratt, I., W. Anderson, and D. Crowley. 2013. Brominated and Fluorinated Organic Pollutants in the Breast Milk of First-Time Irish Mothers: Is There a Relationship to Levels in Food? Food Additives and Contaminants Part A - Chemistry Analysis Control Exposure and Risk Assessment, 30(10), 1788-1798. Rawn, D. F. K., J. J. Ryan, A. R. Sadler, and et al. 2014. Brominated Flame Retardant Concentrations in Sera from the Canadian Health Measures Survey (Chms)from 2007 to 2009. Environment International, 63(1), 26-34. Reistad, T., F. Fonnum, and E. Mariussen. 2006. NeurotoxicityofthePentabrominatedDiphenyl Ether Mixture, De-71, and HBCD (HBCD) in Rat Cerebellar Granule Cells in Vitro. . Arch. Toxicol., 80(11), 785-796. Roberts, C. A., and J. P. Swigert. 1997. Hexabromocyclododecane (HBCD): A 96-Hour Toxicity Test with the Freshwater Alga (Selenastrum Capricornutum). Report# 439A-103. TSCA Section 8D; OTS0573662, DCN 86970000799. Wildlife International Ltd, Easton, MD. Ronisz, D., E. F. Finne, H. Karlsson, and L. Forlin. 2004. Effects of the Brominated Flame Retardants Hexabromocyclododecane (HBCDD), and Tetrabromobisphenol a (TBBPA), on Hepatic Enzymes and Other Biomarkers in Juvenile Rainbow Trout and Feral Eelpout. Aquatic Toxicology, 69(1), 229-245. Saegusa, Y., H. Fujimoto, G. H. Woo, K. Inoue, M. Takahashi, K. Mitsumori, A. Nishikawa, and M. Shibatani. 2009. Developmental Toxicity of Brominated Flame Retardants, Tetrabromobisphenol a and 1,2,5,6,9,10-Hexabromocyclododecane, in Rat Offspring after Maternal Exposure from Mid-Gestation through Lactation. Reproductive Toxicology, 28, 456-467. Sahlstrom, L., U. Sellstrom, and C. A. de Wit. 2012. Clean-up Method for Determination of Established and Emerging Brominated Flame Retardants in Dust. Analytical and Bioanalytical Chemistry, 404(2), 459-466. Sanders, J. M., G. A. Knudsen, and L. S. Birnbaum. 2013. The Fate of B- Hexabromocyclododecane in Female C57bl/6 Mice. Toxicological Sciences, 134(2), 251- 257. Page 55 of 97 ------- SGP (Scientific Guidance Panel). 2014. Biomonitoring California Priority Chemicals. State of California Environmental Protection Agency, Office of Environtal Health Hazard Assessment, Sacramento, California. http://www.biomonitoring.ca.gov/chemicals/chemicals-biomonitored-california (accessed on January 23, 2015). Shi, Z., Y. Jiao, Y. Hu, Z. Sun, X. Zhou, J. Feng, J. Li, and Y. Wu. 2013. Levels of Tetrabromobisphenol a, Hexabromocyclododecanes and Polybrominated Diphenyl Ethers in Human Milk from the General Population in Beijing, China. Science of the Total Environment, 452-453,10-18. Shoeib, M., T. Harner, G. M. Webster, E. Sverko, and Y. Cheng. 2012. Legacy and Current-Use Flame Retardants in House Dust from Vancouver, Canada. Environmental Pollution (Series A). Ecological and Biological, 169,175-182. Siebel-Sauer, D. B., and D. Bias (BASF Corporation. Submitted to the U.S. Environmental Protection Agency under TSCA Section 8D). 1990. Algal Growth Inhibition Test with Cover Letter Dated 031290. OTS0522954. BASF Corporation, Wyandotte, Ml. Smolarz, K., and A. Berger. 2009. Long-Term Toxicity of Hexabromocyclododecane (HBCDD) to the Benthic Clam Macoma Balthica (L.) from the Baltic Sea. Aquatic Toxicology, 95(3), 239-247. SRI (SRI Research Institute). 1990. In Vitro Microbiological Mutagenicity Studies of Four Ciba- Geigy Corporation Compounds (Final Report). . 86-900000262; NTIS No. OTS0523254. With test data and cover letter. Submitted under TSCA Section 8D., Washington, DC. Stapleton, H. M., J. G. Allen, S. M. Kelly, A. Konstantinov, S. Klosterhaus, D. Watkins, M. D. McClean, and T. F. Webster. 2008a. Alternate and New Brominated Flame Retardants Detected in U.S. House Dust. Environmental Science & Technology, 42(18), 6910-6916. Stapleton, H. M., N. G. Dodder, J. R. Kucklick, C. M. Reddy, M. M. Schantz, P. R. Becker, F. Gulland, B. J. Porter, and S. A. Wise. 2006. Determination ofHBCD, PBDEs and Meo-Bdes in California Sea Lions (Zalophus Californianus) Stranded between 1993 and 2003. Marine Pollution Bulletin, 52, 522-531. Stapleton, H. M., S. Klosterhaus, S. Eagle, J. Fuh, J. D. Meeker, A. Blum, and T. F. Webster. 2009. Detection ofOrganophosphate Flame Retardants in Furniture Foam and U.S. House Dust. Environmental Science &Technology, 43(19), 7490-7495. Stapleton, H. M., J. Misenheimer, K. Hoffman, and T. F. Webster. 2014. Flame Retardant Associations between Children's Handwipes and House Dust. Chemosphere, 116, 54-60. Page 56 of 97 ------- Stockholm Convention (Stockholm Convention on Persistent Organic Pollutants). 2010. Draft Risk Profile on Hexabromocyclododecane Chatelaine, Switzerland. Stockholm Convention (Stockholm Convention on Persistent Organic Pollutants). 2013. Conference of the Parties Decision 6/13. Chatelaine, Switzerland. http://chm.pops.int/default.aspx. Suh, K. W. 2000. Foamed Plastics. In Seidel, A., and M. Bickford, Kirk-Othmer Encyclopedia of Chemical Technology (pp. 1-45). John Wiley & Sons, Inc., Online. http://dx.doi.org/10.1002/0471238961.06150113192108.a01. Szabo, D. T., J. J. Diliberto, H. Hakk, and et al. 2010. Toxicokinetics of the Flame Retardant HBCD Gamma: Effect of Dose, Timing, Route, Repeated Exposure, and Metabolism. Toxicological Sciences, 117(2), 282-293. Szabo, D. T., J. J. Diliberto, H. Hakk, J. K. Huwe, and L S. Birnbaum. 2011b. Toxicokinetics of the Flame Retardant Hexabromocyclododecane Alpha: Effect of Dose, Timing, Route, Repeated Exposure, and Metabolism. Toxicological Sciences, 121(2), 234-244. Takigami, H., G. Suzuki, Y. Hirai, and S. Sakai. 2008. Transfer of Brominated Flame Retardants from Components into Dust inside Television Cabinets. Chemosphere, 73,161-169. Takigami, H., G. Suzuki, Y. Hirai, and S. Sakai. 2009a. Brominated Flame Retardants and Other Polyhalogenated Compounds in Indoor Air and Dust from Two Houses in Japan. Chemosphere, 76, 270-277. TemaNord (Nordic Council of Ministers). 2008. Hexabromocyclododecane as a Possible Global POP. ISBN 978-92-893-1665-1. Copenhagen, Denmark. http://www.unece.org/env/documents/2008/EB/EB/Norway%20HBCDD%20dossier.pdf. Thomas, S., H. O. Kreuger, and T. Z. Kendall. 2003a. Hexabromocyclododecane (HBCD): A Prolonged Sediment Toxicity Test with Hyalella Azteca Using Spiked Sediment with 2% Total Organic Carbon. Reporttf 439A-119B, 103 pp. TSCA Section FYI-1103-01472; DCN 84040000010, pg 150. Wildlife International Ltd, Easton, MD. http://www.epa.gov/opptintr/tsca8e/pubs/8ehq/2003/nov03/fyi 1103 01472a.pdf. Thomas, S., H. O. Kreuger, and T. Z. Kendall. 2003b. Hexabromocyclododecane (HBCD): A Prolonged Sediment Toxicity Test with Hyalella Azteca Using Spiked Sediment with 5% Total Organic Carbon. Final Reporttf 439A-120, 103 pp. TSCA Section FYI-1103-01472; DCN 84040000010, pg 253. Wildlife International Ltd, Easton, MD. http://www.epa.gov/opptintr/tsca8e/pubs/8ehq/2003/nov03/fvi 1103 01472a.pdf. Thomsen, C, P. Molander, H. L. Daae, and et al. 2007. Occupational Exposure to HBCD at an Industrial Plant. Environmental Science &Technology, 41(15), 5210-5216. Page 57 of 97 ------- Thuresson, K., J. A. Bjorklund, and C. A. De Wit. 2012. Tri-Decabrominated Diphenyl Ethers and Hexabromocyclododecane in Indoor Air and Dust from Stockholm Microenvironments 1: Levels and Profiles. Science of the Total Environment, 414, 713-721. Tomy, G. T., K. Pleskach, S. H. Ferguson, J. Hare, G. Stern, G. Macinnis, C. H. Marvin, and L. Loseto. 2009. Trophodynamics of Some PFCs and BFRs in a Western Canadian Arctic Marine Food Web. Environmental Science &Technology, 42(11), 4076-4081. Tomy, G. T., K. Pleskach, T. Oswald, T. Halldorson, P. A. Helm, G. Macinnis, and C. H. Marvin. 2008. Enantioselective Bioaccumulation of Hexabromocyclododecane and Congener- Specific Accumulation ofBrominated Diphenyl Ethers in an Eastern Canadian Arctic Marine Food Web. Environmental Toxicology and Chemistry, 42(10), 3634-3639. Tue, N. M., S. Takahashi, G. Suzuki, T. Isobe, P. H. Viet, Y. Kobara, N. Seike, G. Zhang, A. Sudaryanto, and S.Tanabe. 2013. Contamination of Indoor Dust and Air by Polychlorinated Biphenyls and Brominated Flame Retardants and Relevance of Non- Dietary Exposure in Vietnamese Informal E-Waste Recycling Sites. Environment International, 51,160-167. Ueno, D., M. Alaee, C. Marvin, D. C. G. Muir, G. Macinnis, E. Reiner, P. Crozier, V. I. Furdui, A. Subramanian, G. Fillman, P. K. S. Lam, G. J. Zheng, M. Muchtar, H. Razak, M. Prudente, K. Chung, and S. Tanabe. 2006. Distribution and Transportability of Hexabromocyclododecane (HBCD) in the Asia-Pacific Region Using Skipjack Tuna as a Bioindicator. Environmental Pollution, 144(1), 238-247. UNEP (United Nations Environment Programme). 2010. Hexabromocyclododecane Draft Risk Profile. Europe. http://chm.pops.int/Convention/POPsReviewCommittee/hrPOPRCMeetings/POPRC5/P OPRC5Followupcommunications/HBCDInvitationforcommentsondraftRP/tabid/742/lang uage/en-US/Default.aspx.. USITC (US International Trade Commision). 2013. Interactive Tariff and Trade Dataweb. US Department of Commerce and US International Trade Commision, Washington, DC. http://dataweb.usitc.gov/ (accessed on January 14, 2014). van den Eede, N., A. C. Dirtu, N. AN, H. Neels, and A. Covaci. 2012. Multi-Residue Method for the Determination of Brominated and Organophosphate Flame Retardants in Indoor Dust. Talanta, 89, 292-300. van der Ven, L. T., T. van de Kuil, P. E. Leonards, and et al. 2009. Endocrine Effects of HBCD (HBCD) in a One-Generation Reproduction Study in Wistar Rats. Toxicology Letters, 185(1), 51-62. Page 58 of 97 ------- van der Ven, L. T., A. Verhoef, T. Van de Kuil, W. Slob, P. E. G. Leonards, T. J. Visser, T. Hamers, M. Merlin, H. Hakansson, H. Olausson, A. H. Piersma, and J. G. Vos. 2006. A 28-Day Oral Dose Toxicity Study Enhanced to Detect Endocrine Effects of Hexabromocyclododecane in WistarRats. Toxicological Sciences, 94(2), 281-292. Veith, G. D., D. L. Defoe, and B. V. Bergstedt. 1979. Measuring and Estimating the Bioconcentration Factor of Chemicals in Fish. Journal of The Fisheries Research Board of Canada, 36, 1040-1048. Venkatesan, A. K., and R. U. Halden. 2014. Wastewater Treatment Plants as Chemical Observatories to Forecast Ecological and Human Health Risks of Manmade Chemicals. Scientific Reports, 4(3731), 1-7. Walsh, G. E., M. J. Yoder, L. L. Mclaughlin, and E. M. Lores. 1987. Responses of Marine Unicellular Algae to Brominated Organic Compounds in Six Growth Media. Ecotoxicology and Environmental Safety, 14, 215-222. Wang, T., S. Han, T. Ruan, Y. Wang, J. Feng, and G. Jiang. 2013. Spatial Distribution and Inter- Year Variation of Hexabromocyclododecane (HBCD) and Tris-(2,3-Dibromopropyl) Isocyanurate (Tbc) in Farm Soils at a Peri-Urban Region. Chemosphere, 90,182-187. Weil, E. D., and S. V. Levchik. 2009. Flame Retardants for Plastics and Textiles: Practical Applications. Cincinnati, OH: Hanser Publications. http://www.hanser- elibrary.com/isbn/9783446416529. Weiss, J., E. Walling, A. Axmon, and et al. 2006. Hydroxy-Pcbs, PBDEs, and Hbcdds in Serum from an Elderly Population of Swedish Fishermen's Wives and Associations with Bone Density. Environmental Science &Technology, 40(20), 6282-6289. WSDE (Washington State Department of Ecology). 2013. Chemicals of High Concern to Children. Olympia, Washington, http://www.ecy.wa.gov/programs/swfa/cspa/chcc.html (accessed on January 23, 2015). WWF (World Wildlife Fund). 2004. Chemical Check Up: An Analysis of Chemicals in the Blood of Members of the European Parliament. http://www.panda.org/downloads/europe/checkupmain.pdf. Xu, J., Y. Zhang, C. CGuo, Y. He, L. Li, and W. Meng. 2013. Levels and Distribution of Tetrabromobisphenol a and Hexabromocyclododecane in Taihu Lake, China. Environmental Toxicology and Chemistry, 32(10), 2249-2255. Yu, C. C., and Y. H. Atallah. 1980. Pharmacokinetics of HBCD in Rats Vesicol Chemical Corporation, Rosemont, II. Page 59 of 97 ------- Zeiger, E., B. Anderson, S. Hawarth, T. Lawlor, K. Mortelmans, and W. Speck. 1987. Salmonella Mutagenicity Tests: III. Results from the Testing of 255 Chemicals. Environmental Mutagenesis, 9(Suppl. 9), 1-110. Zeller, H., and P. Kirsch. 1969. Hexabromocyclododecane: 28-Day Feeding Trials with Rats with Cover Letter Dated 031290. OTS0522939. BASF Corporation, Parsippany, NJ. Zeller, H., and P. Kirsch. 1970. Hexabromocyclododecane: 90-Day Feeding Trials with Rats. BASF Unpublished Report, Parsippany, NJ. (as cited in BFRIP, 2001). Zhang, H., Y.-Y. Kuo, A. C. Gerecke, and J. Wang. 2012. Co-Release of Hexabromocyclododecane (HBCD) and Nano- and Microparticlesfrom Thermal Cutting of Polystyrene Foams. Environmental Science & Technology, 46(20), 10990-10996. Zhang, X., F. Yang, Y. Xu, T. Lisao, S. Song, and J. Wang. 2008. Induction of Hepatic Enzymes and Oxidcative Stress in Chinese Rare Minnow (Gobiocypris Rams) Exposed to Waterborne Hexabromocyclododecane (HBCD). Aquatic Toxicology, 86, 4-11. Zhang, Y., Y. Ruan, H. Sun, L. Zhao, and Z. Can. 2013. Hexabromocyclododecanes in Surface Sediments and a Sediment Core from Rivers and Harbor in the Northern Chinese City of Tianjin. Chemosphere, 90,1610-1616. Zhou, D., Y. Wu, X. Feng, Y. Chen, Z. Wang, T. Tao, and D. Wei. 2014. Photodegradation of Hexabromocyclododecane (HBCD) by Fe(lll) Complexes/^O 2 under Simulated Sunlight. Environmental Science Pollution Research, 21(21), 6228-6233. Zhou, D. N., L. Chen, F. Wu, J. Wang, and F. Yang. 2012. Debromination of Hexabromocyclododecane in Aqueous Solutions by UV-C Irradiation. Fresenius Environmental Bulletin, 21(1), 107-111. Page 60 of 97 ------- APPENDICES Appendix A Regulatory and Assessment History Table_Apx A-l: Regulatory and Assessment History of HBCD8 COUNTRY/ORGANIZATION UNITED STATES ASSESSMENT Environmental Protection Agency • Office of Environmental Information - Proposed HBCD for listing to the Toxic Release Inventory (TRI) Program (2014). For current list of chemicals see: http://www2.epa.gov/toxics-release-inventorv-tri-program/tri-listed-chemicals • Office of Research and Development - Draft Toxicological Review Scoping Document to support an IRIS assessment (http://www.ecv.wa.gov/programs/wq/swqs/IRISAgendaChemicals.pdf) • Office of Pollution Prevention and Toxics (OPPT) - Flame retardant alternatives to hexabromocyclododecane (2014) (http://www.epa.gov/dfe/pubs/proiects/hbcd/hbcd-full-report-508.pdf) • OPPT - Proposed SNUR (Mar 2012) to designate manufacture or processing of HBCD for use as a flame retardant in consumer textiles as a significant new use • OPPT Action Plan for HBCD (2010) (all congeners; http://www.epa.gov/oppt/existingchemicals/pubs/actionplans/RIN2070- AZ10 HBCD%20action%20plan Final 2010-08-09.pdf) • OPPT Risk Based Prioritization (RBP) including Hazard Characterization (2008) (http://www.epa.gov/chemrtk/hpvis/rbp/HBCD.3194556.Web.RBP.31308.pdf) for CASRNs 3194-55-6 and 25637-99-4 • OPPT - CASRN 3194-55-6 (2001) High Production Volume Challenge Program test plan and robust summaries submission (http://www.epa.gov/chemrtk/pubs/summaries/cyclodod/cl3459cv.pdf) Occupational Safety and Health Administration (OSHA) • No occupational exposure limits (OSHA PEL, NIOSH REL, ACGIH TLV) are established (www.osha.gov) Consumer Product Safety Commission (CPSC) • CPSC staff exposure and risk assessment of flame retardant chemicals in residential upholstered furniture (2001) (http://www.cpsc.gov/en/) • CPSC staff preliminary risk assessment of flame retardant (FR) chemicals in upholstered furniture foam (2006) (http://www.cpsc.gov/en/) • Quantitative assessment of potential health effects from the use of fire retardant (FR) chemicals in mattresses (2006) (http://www.cpsc.qov/en/) State -California • HBCD is listed as an informational initial candidate chemical under California's Safer Consumer Products regulations. (DTSC, 2010) (http://www.dtsc.ca.gov/SCP/ChemList.cfm) • HBCD is listed on the state's Proposition 65 list because it is known to cause cancer or birth defects or other reproductive harm (OEHHA, 2007, 2014). (http://oehha.ca.qov/) (http://oehha.ca.qov/prop65/prop65 Nst/files/P65sinqle050214.pdf) 8 The risk assessment conclusions summarized in the table are selected conclusions from the reports that address the pertinent US exposure scenarios and should not be construed as EPA's conclusions. EPA refers the reader to the full reports for detailed explanations of the context of the conclusions reached in all risk assessments. ------- COUNTRY/ORGANIZATION CANADA EUROPEAN UNION AUSTRALIA ASSESSMENT • California lists HBCD as a designated and priority chemical for biomonitoring. However, California has not yet started biomonitoring HBCD (SGP, 2014). (httD://www.biomonitorina.ca.aov/chemicals/chemicals-biomonitored-california) State - Maine • Maine classifies HBCD as a chemical of high concern (DEP, 2013). (httD://www.maine.aov/deD/safechem/hiahconcern/) State - Minnesota • Minnesota classifies HBCD as a chemical of high concern (MDH, 2013). (httD://www.health. state. mn.us/divs/eh/hazardous/toDics/toxfreekids/chclist/mdhchc2013.Ddf) State - Oregon • The Oregon Department of Environmental Quality lists HBCD as a priority persistent pollutant (DEQ, 2010a, 2011). (http://www.deq.state.or.us/wq/SB737/docs/LeaRpAtt20100601.pdf) (http://www.deq.state.or.us/wq/SB737/) • Oregon posts use, exposure pathways and release data for HBCD under this program (DEQ, 2010b). (httD://www.dea.state.or.us/wa/SB737/docs/LeaRDAtt420100601.Ddf) State - Washington • Washington classifies HBCD as a chemical of high concern to children (WSDE, 2013). (http://www.ecv.wa.qov/proqrams/swfa/cspa/chcc.html) • HBCD meets Canada's regulatory criteria for persistence and bioaccumulation potential. (http://www.ec.gc.ca/ese-ees/default. asp?lang=En&n=7882C148-l#a4) • On November 1, 2012, Canada added HBCD to Schedule 1 of CEPA 1999 (Virtual Elimination List). Proposed risk reduction measures would prohibit the manufacture, import, use, sale, and offer for sale of HBCD and products containing HBCD. (http://www.ec.gc.ca/lcpe-cepa/eng/regulations/detailReg.cfm?intReg=82) • Health Canada (Health CA) and Environment Canada (ENVCA) have published a screening- level assessment, (http://www.ec.gc.ca/ese-ees/default. asp?lang=En&n=7882C148-l#a4) • HBCD is listed as a substance of very high concern (SVHC) and it is also listed under Annex XIV (Authorisation list) of European Union's REACH. After August 21, 2015, only persons with approved authorization applications may continue to use the chemical. (http://echa.europa.eu/candidate-list-table) (http://echa.europa.eu/documents/10162/13640/prioritisation hbccd en.pdf) • The European Chemicals Agency (ECHA) is currently considering two applications to authorize the use of HBCD: (i) formulation of flame retarded expanded polystyrene (EPS) to solid unexpanded pellets using HBCD as the flame retardant additive (for onward use in building applications); and (i) manufacture of flame retarded expanded polystyrene (EPS) articles for use in building applications. (http://echa.europa.eu/documents/10162/18584504/afa opinion hbcdd use 2 en.pdf) • HBCD is recommended to be reviewed for the ED Directive's list of banned substances under the restriction of hazardous substances (RoHS) in electrical and electronic equipment. (http://ec.europa.eu/environment/waste/weee/pdf/hazardous substances report.pdf) • The WEEE (Waste Electrical and Electronic Equipment) directive in the European Union requires the separation of plastics containing brominated flame retardants prior to recycling, (http://ec.europa.eu/environment/waste/weee/index en. htm) • The European Commission published a European Union Risk Assessment Report (EC, 2008) on HBCD. (http://ec.europa.eu/health/ph risk/com mittees/04 scher/docs/scher o 093.pdf) • HBCD is listed as a Priority Existing Chemical, (http://www.nicnas.gov.au/chemical- information/pec-assessments) Page 62 of 97 ------- COUNTRY/ORGANIZATION JAPAN STOCKHOLM CONVENTION ON PERSISTENT ORGANIC POLLUTANTS (POPs) ORGANISATION FOR ECONOMIC CO- OPERATION AND DEVELOPMENT (OECD) ASSESSMENT • Australia (National Industrial Chemicals Notification and Assessment Scheme) has prepared a risk assessment for HBCD. (http://www.nicnas.aov.au/Publications/CAR/PEC/PEC34/HBCD Report June 2012 PDF.pdf) • HBCD is subject to mandatory reporting requirements in Japan under the Chemical Substances Control Law (CSCL), specifically Japan requires type III monitoring for all substances that may interfere with the survival and/or growth of flora and fauna. (http://www.meti.go.ip/policy/chemical management/english/cscl/files/about/150408Progres.pdf) • In May 2013, HBCD was added to the United Nation's Stockholm Convention list of Persistent Organic Pollutants. The chemical is scheduled to be eliminated by November 2014 with specific exemptions for production and uses in expanded or extruded polystyrene building insulation. As required by the convention, parties that use these exemptions must register with the secretariat and the exemptions will expire in November 2019. (http://chm.pops.int/default.aspx) • Published SIDS Initial Assessment Profile (SIAP) and SIAR in 2007 for CASRNs 3194-55-6 and 25637-99-4. (http://webnet.oecd.org/HPV/UI/SIDS Details.aspx?key=39a31bc9-3719- 4c55-a7d4-a8bbdd9afe04&idx=0) (http://webnet.oecd. org/HPV/UI/handler.axd?id=ea58acll-e090-4b24-b281- 200ae351686c) Page 63 of 97 ------- Appendix B Uses Supplement Tables Table_Apx B-l: 2012 CDR Production Data (Data Reported for 2011) CASRN 25637-99-4 3194-55-6 Manufacturing Site CBI CBI BASF Corporation 100 Campus Drive Florham Park, NJ 07932 Albemarle Corporation 1550 Hwy. 371 W. Magnolia, AR 71753 The Dow Chemical Company 2020 Dow Center Midland, Ml 48674 Domestic Manufacturing CBI CBI ND CBI ND Imported ND ND CBI ND CBI Volume Exported (Ibs) CBI 0 CBI CBI 0 Volume Used on the Site (Ibs)1 0 0 CBI CBI N/A 2010 Past Production Volume (import+ manufacture) CBI CBI CBI CBI CBI 2011 National Production Volume (Ibs/yr) Withheld Withheld ^he total volume (domestically manufactured and imported) of the chemical used at the reporting site. This number represents the volume of the chemical that did not leave the manufacturing site. CBI = Confidential Business Information ND = No Data; the company did not provide the requested information. N/A = Not Applicable; the imported chemical was never physically at the site. "Withheld" in the CDR public database indicates that the national production volume of a chemical was unable to be aggregated in order to protect CBI claims. ------- Table_Apx B-2: Historic IUR and CDR Production Volumes CASRN 25637-99-4 3194-55-6 Year 1986 10K-500K XLM-10M 1990 No Reports >1M-10M 1994 No Reports >10M - 50M 1998 10K - 500K >10M - 50M 2002 10K - 500K >10M - 50M 2006 No Reports 10 to < 50M 2010 CBI CBI 2011 Withheld Withheld ^The total volume (domestically manufactured and imported) of the chemical used at the reporting site. This number represents the volume of the chemical that did not leave the manufacturing site. CBI = Confidential Business Information "Withheld" in the CDR public database indicates that the national production volume of a chemical was unable to be aggregated in order to protect the CBI claims. Table_Apx B-3: Summary of 2011 CDR Production Volume and Use Information CAS Number 25637-99-4 Industrial Sector Reported in CDR Plastics Material and Resin Manufacturing Construction Description of Industrial Use Flame retardant in electrical and electronic equipment Flame retardant in insulation boards Commercial or Consumer Product Category Plastic and Rubber Products not Covered Elsewhere (Commercial and Consumer use) Building/Construction Materials Not Covered Elsewhere (Commercial and Consumer use) Building/Construction Materials Not Covered Elsewhere (Commercial use) Potential End Product • Electric housings for VCR • Electrical and electronic equipment (e.g., distribution boxes for electrical lines) • Video cassette housings • Construction, insulation boards (packaging material) • Insulation boards (against cold or warm) of transport vehicles (e.g., lorries and caravans) • Insulation boards in building constructions, e.g. houses' walls, cellars and indoor ceilings and "inverted roofs" (outdoor) • Insulation boards against frost heaves of road and railway embankments 2011 PV CBI CBI CBI Approximate % of 2011 National PV CBI CBI 100 Page 65 of 97 ------- CAS Number 3194-55-6 Industrial Sector Reported in CDR Utilities Description of Industrial Use Flame retardant in insulation boards Commercial or Consumer Product Category Building/Construction Materials Not Covered Elsewhere (Consumer use) Building/Construction Materials Not Covered Elsewhere (Commercial use) Potential End Product • Construction, insulation boards (packaging material) • Insulation boards in building constructions, e.g., houses' walls, cellars and indoor ceilings and "inverted roofs" (outdoor) • Construction, insulation boards,(packaging material) • Insulation boards (against cold or warm) of transport vehicles (e.g., lorries and caravans) • Insulation boards in building constructions e.g. houses' walls, cellars and indoor ceilings and "inverted roofs" (outdoor) • Insulation boards against frost heaves of road and railway embankments 2011 PV CBI CBI Approximate % of 2011 National PV 100 50 Note: 1) Plastic and rubber products with consumer/commercial categories in the CDR data include: • Food packaging • Toys, playground, and sporting equipment 2) Building and construction materials with consumer/commercial categories in the CDR data include: • Building/construction materials - wood and engineered wood products CBI = Confidential Business Information PV= Production Volume Page 66 of 97 ------- Table_Apx B-4: Uses of HBCD as Listed in the 2008 EU Risk Assessment Material1 Use/ Function1 Percent of HBCD Production Volume2 End Products1 Ongoing Use in the United States (US)? EPS Insulation 45% Construction, insulation boards (packaging material) Packaging material (minor use and not in food packaging) Insulation boards (against cold or warm) of transport vehicles (e.g., lorries and caravans) Insulation boards in building constructions e.g. houses' walls, cellars and indoor ceilings and "inverted roofs" (outdoor) Insulation boards against frost heaves of road and railway embankments Yes; based on use in construction insulation boards in 2012 CDR data2 Unknown; it is unclear if this is an ongoing use in the US, and uses of polystyrene foam in consumer products generally do not require the use of a flame retardant3 Possibly; based on use in insulation boards in 2012 CDR data2 Yes; based on use in construction insulation boards in 2012 CDR data2 Possibly; based on use in insulation boards in 2012 CDR data2 XPS Insulation 51% Construction, insulation boards Insulation boards (against cold or warm) of transport vehicles (e.g. lorries and caravans) Insulation boards in building constructions e.g. houses' walls, cellars and indoor ceilings and "inverted roofs" (outdoor) Insulation boards against frost heaves of road and railway embankments Yes; based on use in construction insulation boards in 2012 CDR data2 Possibly; based on use in insulation boards in 2012 CDR data2 Yes; based on use in construction insulation boards in 2012 CDR data2 Possibly; based on use in insulation boards in 2012 CDR data2 HIPS Electrical and electronic parts 2% Electric housings for VCR Electrical and electronic equipment, e.g., distribution boxes for electrical lines Video cassette housings Possibly, based on use in electronic plastics in 2012 CDR2 Possibly, based on use in electronic plastics in 2012 CDR2 Possibly, based on use in electronic plastics in 2012 CDR2 Page 67 of 97 ------- Material1 Use/ Function1 Percent of HBCD Production Volume2 End Products1 Ongoing Use in the United States (US)? Polymer dispersion on cotton or cotton/ synthetic blends Textile coating agent 2% Upholstery fabric Bed mattress ticking Flat and pile upholstered furniture (residential and commercial furniture) Upholstery seating in transportation Draperies, and wall coverings Interior textiles e.g. roller blinds Automobile interior textiles Historic Use in the US4 Historic Use in the US4 Historic Use in the US4 Possibly, based industry response to SNUR2 Historic Use in the US4 Historic Use in the US4 Possibly, based industry response to SNUR2 Note: The uses in this table describe recorded HBCD applications for both the US and other countries. Given that HBCD will be phased out internationally under REACH and the Stockholm Convention, it is unclear to what extent HBCD is currently used in these applications outside of the US. Sources: 1) EC, 2008 2) EPA, 2012a 3) EPA, 2014 4) EPA, 2012e Page 68 of 97 ------- Appendix C Exposure Supplement Tables Table_Apx C-l: 2012 CDR Data (Data Reported for 2011) for Release Assessment Table Line Number 1 2 3 4 5 6 7 Manufacture / Import Site Identity CBI BASF, Florham Park, NJ Albemarle Corporation, Magnolia, AR The Dow Chemical Company, Midland, Ml Max Concentration and Physical Form 90%+; Dry Powder or Other Solid lto<30%; Pellets / Large Crystals 90%+; Dry Powder 60 to < 90%; Pellets/ Large Crystals Industrial Processing/ Use Processing Use Processing-incorporation into formulation, mixture, or reaction product Processing-incorporation into article Processing-incorporation into article Processing-incorporation into article Processing-incorporation into article Processing-incorporation into formulation, mixture, or reaction product Processing Sector Plastics Material and Resin Manufacturing Construction Construction Plastics Material and Resin Manufacturing Utilities Utilities Function Flame retardants Flame retardants Other Other Flame retardants Flame retardants %PV 50 50 CBI CBI NKRA NKRA Number of Sites <10 <10 10 to 24 25 to 99 10 to 24 10 to 24 Commercial/ Consumer Use Use Product Category Building/ Construction Materials Not Covered Elsewhere Building/ Construction Materials not covered elsewhere Plastic and Rubber Products not covered elsewhere Building/ Construction Materials not covered elsewhere Not Reported CBI = Confidential Business Information NKRA = Not Known or Reasonably Ascertainable PV = Production Volume ------- Table_Apx C-2: Compilation of Release Factors and Other Release-Related Information in Various Risk Assessments Operation (Manufacture or Processing Step) Manufacture of EPS resin beads that contain HBCD (also referred to as "formulation of EPS compound" (EC, 2008) or "compounding raw HBCD into resins" (NICNAS, 2012)) Manufacture of EPS that contains HBCD (also referred to as "industrial use of EPS compound" (EC, 2008) or "conversion or processing of polymeric resin into EPS foam products" (NICNAS, 2012)) Compounding of Polystyrene Resin to Produce XPS Masterbatch containing HBCD (also referred to as "formulation of XPS compound for the manufacture of Reference EC (2008) NICNAS (2012) EC (2008) NICNAS (2012) EC (2008) Release Media water air water air water air water air water air Loss Factor (Emission Factor) Value 7.6E-06 7.3E-06 6.8E-03 2.5E-04 3.0E-05 3.0E-05 1.6E-03 1.6E-03 7.4E-06 7.3E-06 Description This is the emission factor for a generic site and was determined based on the 90th percentile of measurements of concentration in the effluent from the on-site sewer treatment plants of 9 of the total of 13 sites. This is the emission factor for a generic site and is equal to the maximum emission factor calculated from site-specific data that pertain to three processing steps including formulation of EPS and XPS compounds. These values are reasonable worst case values that are the sums of emission factors for handling and compounding solid flame retardants with particle sizes <40 urn that are also of relatively high volatility as reported in OECD (2004b). These are the emission factors for the generic site and are reported in OECD (2004b) as the emission factors for organic flame-retardants of relatively low volatility in partially open plastics conversion processes. These emission factors are weighted averages of emission factors for organic flame of relatively high volatility in closed plastics conversion processes that are reported in OECD (2004b). The emission factors that were averaged include values for high process temperature and low plastics production volume. This is the emission factor for a generic site and is equal to the maximum of emission factors calculated from site- specific data for three sites. This is the emission factor for a generic site and is equal to the maximum emission factor calculated from site-specific data that pertain to three processing steps including formulation of EPS and XPS compounds. Number of Release Days Value 300 150 300 200 300 Description This is the value is for the generic site and was determined in accordance with the general risk assessment guidance of the EU. Site-specific values are in the range of 61 to 350 with the exception of one value that is equal to 1. This is a site-specific value. This is the value for the generic site and was determined in accordance with the general risk assessment guidance of the EU. This is an assumed value. This is the value for the generic site and was determined in accordance with the general risk assessment guidance of the EU. Page 70 of 97 ------- Operation (Manufacture or Processing Step) flame retarded XPS" (EC, 2008) Manufacture of XPS using XPS Masterbatch containing HBCD Reference Environment CAand Health CA (2011) EC (2008) Release Media water water air Loss Factor (Emission Factor) Value 6.6E-03 2.6E-05 5.8E-05 Description This is the emission factor for a generic site and is the sum of emission factors for handling and compounding organic solid flame retardants with particle sizes <40 urn that are of relatively medium volatility as reported in OECD (2004b). These are the emission factors for the generic site and are equal to the maxima of the emission factors that were derived from site-specific data but were not explicitly reported. EPA calculated these factors, each of which is equal to the ratio of total annual releases from 13 sites for which site-specific data was not reported and the total annual processing rate for these sites. Number of Release Days Value 60 or 200 1 300 Description These values are for generic sites, are functions of the processing rate, and were determined in accordance with the general risk assessment guidance of the EU. This value was assumed as a reasonable worst case for the number of release days from a generic site. Site-specific values are in the range of 1 to 15. The number of release days for the generic site is not reported. Site- specific values are in the range of 15 to 300. Page 71 of 97 ------- Table_Apx C-3: Preliminary Values of Input Variables for Calculation of the Range of Release Rates from Manufacturing Sites or the Processing Sites of Each Processing Step Operation (Manufacture or Processing Step) manufacture of HBCD manufacture of EPS resin beads manufacture of EPS manufacture of XPS using HBCD powder compounding of polystyrene resin to produce XPS masterbatch containing HBCD manufacture of XPS using XPS masterbatch containing HBCD Processing Rates as Fractions of the HBCD Manufacturing Rate1 Not Applicable 0.45 0.45 0.51 0.51 0.51 Manufacturing or Processing Rate (kg/yr)2 lower limit of range 4.540.E+06 2.043.E+06 2.043.E+06 2.316.E+06 2.316.E+06 2.316.E+06 upper limit of range 2.290.E+07 1.031. E+07 1.031. E+07 1.168.E+07 1.168.E+07 1.168.E+07 Number of Sites3 minimum maximum 3 1 5 6 10 10 4 15 18 29 24 Number of Release Days4 250 250 250 1 (water) 250 (air) 250 1 (water) 250 (air) The Minimum and Maximum Values of the Compiled5 and Calculated6 Loss Factors for Each Medium of Release (kg of HBCD released per kg of HBCD manufactured or processed) water min 1.2E-07 7.6E-06 3.0E-05 l.OE-05 7.4E-06 max 4.0E-04 6.8E-03 1.6E-03 l.OE-05 6.6E-03 2.6E-05 air min max 3.3E-07 6.8E-04 7.3E-06 2.5E-04 3.0E-05 1.6E-03 7.3E-06 7.3E-06 5.8E-05 Note: 1) The share of production volume that is processed to manufacture of EPS or XPS is reported in Table_Apx B-4. The share of production volume that is processed to manufacture of XPS using HBCD powder or HBCD masterbatch is unknown and therefore EPA will conservatively assume this share is equal to the total share that is processed to manufacture XPS. Page 72 of 97 ------- 2) EPA estimated the lower-end of the current range of production volumes by summing the lower-ends of the ranges of production volumes for CAS number 25637-99-4 reported in 2002 and CAS number 3194-55-6 reported in 2006 that are given in Table_Apx B-2. EPA estimated the upper-end of the current range of production volume by summing the upper- ends of these two ranges. 3) The number of sites was estimated from data reported in EPA (2014b) and Table_Apx C-l. 4) EPA estimated the number of release days to be equal to 250 assuming each facility has an operation schedule of five days per week and 50 weeks per year, allowing for two-week annual downtime for maintenance. The values of the number of release days given Table_Apx C-2 are of similar magnitude. The exceptions are the values for releases to water from the manufacture of XPS using HBCD powder and XPS using XPS masterbatch; EPA assessed each of these parameters to be 1 day per year in consideration of the data presented in Table_Apx C-2. 5) The compilation of non-site specific release factors is given in Table_Apx C-2. 6) EPA calculated release factors for HBCD manufacturing and the manufacture of XPS using HBCD powder from data reported in EC (2008) because non-site specific release factors are not reported for these two operations. Page 73 of 97 ------- Appendix D Environmental Hazard Study Summaries D-l Persistence in Environmental Media D-l-1 Air and Water HBCD does not absorb light in the UV/Visible frequencies so is not expected to undergo direct photolysis in air or water (Zhou et al., 2014). Kajiwara et al (Kajiwara and Takigami, 2013) studied photolysis of HBCD on textiles (4% by wt.) and reported "no substantial loss of any of the HBCD diastereomers during the entire exposure period (371 days)" confirming that photolytic degradation did not occur.Indirect photolysis with methane as a co-solvent has been observed to result in debromination (Zhou et al., 2012) and indirect photolysis with Fe(lll) and H2O2 also can occur (Zhou et al., 2014). Although HBCD is expected to exist primarily in the particulate phase in the atmosphere, a small percentage is expected to exist in the vapor phase based on its vapor pressure (Bidleman, 1988; CMA, 1997; Covaci et al., 2006). HBCD in the vapor phase can be degraded by reaction with hydroxyl radicals in the atmosphere with an estimated rate constant of 5.01xlO"12 cm3/iTiolecules-sec at 25 °C corresponding to an atmospheric half-life of 2.1 days (EPA, 1993, 2011a). HBCD associated with particulates is expected to be removed from the atmosphere through wet or dry deposition. Removal rates are unknown and the widespread detection of HBCD in air samples and biota from remote locations far from release locations indicate that long-range atmospheric transport occurs (Covaci et al., 2006; EPA, 2010a; Ueno et al., 2006). D-l-2 Soil, Sediment and Sludge Based on an estimated Koc value of 7.6xl04 HBCD is expected to be fairly immobile in soil and to bind strongly to soil, sediment, and suspended organic matter. It may undergo abiotic and microbial degradation while associated with solids. The limited data available show that biodegradation is slow and that anaerobic processes may be faster than aerobic. A soil simulation test was conducted according to OECD TG 307 for commercial HBCD (BFRIP, 2001; EC, 2008). Activated sludge was inoculated with soil and HBCD at a nominal concentration of 34-89 u.g/kg dry weight for 120 days. The disappearance half-life was 63 days in aerobic soil and >120 days in abiotic aerobic controls. In the anaerobic soil, the half-life was 7 days compared to 82 days in abiotic anaerobic controls. A closed bottle screening-level test for ready biodegradability (OECD TG 301D) was performed using an initial HBCD concentration of 7.7 mg/L and an activated domestic sludge inoculum (BFRIP, 2001; EC, 2008). No biodegradation was observed (0% of the theoretical oxygen demand) over the test period of 28 days. Gerecke et al (Gerecke et al., 2006) studied ------- degradation of BFRs, including HBCD, under anaerobic conditions in digested sewage sludge. They reported a half-live for a technical HBCD mixture of 0.66 days. They found no statistically significant enantioselective degradation of alpha-, beta-, or gamma-HBCD. The reported reduction in concentration the HBCD mixture decreased in sterile controls at a rate 50 times slower in incubations under non-sterile conditions. A water-sediment OECD TG 308 study for commercial HBCD with a nominal concentration of 34-89 u.g/kg dry weight found that aerobic half-lives were 11-32 days in aerobic sediments and 30-190 days in abiotic aerobic controls (BFRIP, 2001; EC, 2008). In the anaerobic sediments, disappearance half-lives were 1.1-1.5 days compared to 9.9-10 days in abiotic anaerobic controls. Davis et al (2006) studied degradation of 14C radiolabeled HBCD with sludge, sediment, and soil simulation tests with initial concentrations of HBCD of 3.6-4.2 mg/L in the sludge systems and 3.0-4.7 mg/kg dry weight in the sediment and soil systems. As shown in Table_Apx D-l below, they observed decrease in total initial radioactivity in the viable systems and abiotic controls. Table_Apx D-l: Percent Decrease in Total Initial Radioactivity in Viable Systems and Abiotic Controls During Sludge, Sediment, and Soil Simulation Tests Using 14C-labeled HBCD Compartment Anaerobic digester sludge Aerobic activated sludge Anaerobic freshwater sediment Aerobic freshwater sediment Aerobic soil Viable System 87% 21% 61% 44% 10% Abiotic Control 84% 15% 33% 15% 6% Time Period 60 days 65 days 112-113 days 112 days 112 days Source: (BFRIP, 2001; Davis et al., 2006; EC, 2008) HBCD degradation observed in these tests was attributed to abiotic reductive dehalogenation. Degradation proceeded through a stepwise process to form TETRABROMOCYCLODODECENE, DIBROMOCYCLODODECADIENE, AND 1,5,9-CYCLODODECATRIENE (See Figure_Apx 2.6.4-1). Further degradation of 1,5,9-CYCLODODECATRIENE was not observed. General trends observed were increased HBCD degradation under anaerobic conditions compared to aerobic conditions and slower degradation of a-HBCD compared to the (3- and v-stereoisomers. Page 75 of 97 ------- Br' Br HBCD tetrabromo- cyclododecene dibromo- cyclododecadiene 1,5,9- cyclododecatriene Source: Davis et al., 2006; ECB, 2008 Figure_Apx 2.6.4-1: Stepwise Reductive Dehalogenation of HBCD D-l-3 Water and Wastewater HBCD is not expected to undergo hydrolysis in environmental waters because of its low solubility and lack of hydrolyzable functional groups. Based on the studies described above it is not expected to be readily degradable in WWTPs and is expected to be persistent in surface and groundwater. It may undergo indirect photolysis or abiotic degradation catalyzed by iron or organic matter and can be biodegraded slowly under aerobic and anaerobic conditions. Degradation rates in the environment are expected to be slower than those in lab studies. This is consistent with environmental observations. For example the detection of HBCD at concentrations ranging from 112 to 70,085 u.g/kg dry weight in sediment at locations near a production site in Aycliffe, UK collected two years after the facility was closed down demonstrates the persistence of this substance in the environment (EC, 2008). D-l-4 Bioaccumulation High bioconcentration of HBCD in aquatic organisms has been observed in fish. Veith et al. (Veith et al., 1979) measured a BCF of 18,100 for HBCD in fathead minnows. In a flow-through bioconcentration test, BCF values of 8974 and 13,085 were determined for HBCD in rainbow trout (EC, 2008). These BCF values indicate that HBCD has a very high potential to bioconcentrate in aquatic organisms. The widespread detection of this substance in aquatic organisms also suggests that HBCD bioconcentrates in the environment (Covaci et al., 2006; EC, 2008). Biomagnification of HBCD in the aquatic food web has also been reported based on measurements in invertebrates, fish, birds, and marine mammals, with the highest levels of HBCD measured in seals and porpoises (Covaci et al., 2006; EC, 2008; EPA, 2010a). Higher concentrations of HBCD were measured in eggs from wild peregrine falcons feeding on birds from terrestrial food webs in southwestern Sweden than in the eggs of captive falcons feeding on domestic chickens, indicating that HBCD bioaccumulation in terrestrial food chains may also be important (EPA, 2010a; Lindberg et al., 2004). Bioaccumulation may be isomer specific with Y-HBCD is the dominant stereoisomer present in commercial HBCD formulations (>75%) Page 76 of 97 ------- (Becher, 2005). Similar proportionality is observed with HBCD stereoisomers detected in environmental media (Covaci et al., 2006). However, in living organisms, a-HBCD is the predominant stereoisomer, accounting for 70-90% of all HBCD detected (Covaci et al., 2006). The reason for the difference in HBCD composition found in the environment compared to that found in living organisms is unknown. Possible explanations include different rates of bioconcentration for the a- and y- stereoisomers or different rates of metabolism within organisms (Covaci et al., 2006; EC, 2008). D-2 Toxicity to Aquatic Organisms The toxicity to aquatic organisms has been summarized in several publications (EC, 2008; Environment CA and Health CA, 2011; EPA, 2008a, 2014b; NICNAS, 2012; OECD, 2007). Guideline studies in freshwater fish, daphnia, and green algae mostly reported no lethal or sublethal effects at concentrations approaching saturation (Calmbacher, 1978; EPA, 2014b; Graves and Swigert, 1997a, 1997b; Roberts and Swigert, 1997; Siebel-Sauer and Bias, 1990). D-2-1 Aquatic Plant Toxicity The toxicity to aquatic plants is summarized below and presented in Table_Apx D-2. The study denoted with an asterisk is proposed for use by EPA/OPPT for risk assessment. Those studies not considered adequate for risk assessment by EPA/OPPT are shaded in the table. Green algae (Selanastrum capricornutum) were exposed to nominal test concentrations of HBCD ranging from 1.5 to 6.8 u.g HBCD/L (Roberts and Swigert, 1997). The mean measured concentration of HBCDs (3.7 u.g HBCD/L) at the maximum nominal test concentration of 6.8 u.g HBCD/L (with solvent) was similar to the independently measured water solubility limit (8.6 u.g/L). No effects were observed at any test concentration. Another freshwater aquatic plant study reported that green algae (S. subspicatus) had no effects at nominal test concentrations ranging from 7.8 to 500 mg HBCD/L (Siebel-Sauer and Bias, 1990). However, the actual concentrations of HBCD in test solution are unknown. No effects were observed at the highest tested concentration. Adverse effects observed following exposure were found in studies with the estuarine/marine algae species Skeletonema costatum (Desjardins et al., 2004, 2005; Walsh et al., 1987). Walsh et al. (1987) reported measured 72-hour ECso values in S. costatum ranging from 0.009 to 0.012 mg HBCD/L based on reduced growth rate in five different types of saltwater media. The study also tested two species, Chlorella sp. and Thalassiosira pseudonana, that were found to be less sensitive. Desjardins et al. (2005) further substantiated observed toxicity in S. costatum when a single saturated solution of 0.0545 mg HBCD/L (without a solvent) resulted in 51% growth inhibition after 72 hours of exposure. Desjardins et al. (2004) also reported 19, 21, and 7.3% inhibition of cell density, biomass, and growth rate, respectively, following exposure of S. costatum to 0.041 mg HBCD/L (with a solvent), the only concentration tested, for 72 hours. Page 77 of 97 ------- Table_Apx D-2: Toxicity of HBCD to Aquatic Plants Test Species Green algae (Selanastrum capricornutum) Green algae (S. subspicatus) Algae (S. costatum) Algae (S. costatum) Algae (S. costatum) *Algae (S. costatum) Algae (Thalassiosira pseudonana) Algae (Chlorella Sp.) Fresh/ Salt Water Fresh Fresh Marine Marine Marine Marine Marine Marine Duration 96-hour 96-hour 72-hour 72-hour 72-hour 72-hour 72-hour 72-hour End- point ECso ECso ECso ECso ECso ECso ECso ECso Cone. (mg/L) >0.0037 >500 >0.041 >0.01 0.052 0.009- 0.012 0.05-0.37 >1.5 Test Analysis Static, Measured Static, Nominal Static, Measured Static, Measured Static, Measured Static, Measured Static, Measured Static, Measured Effect Biomass/Growth rate Growth inhibition Growth rate References Roberts and Swigert, 1997 Siebel-Sauer and Bias, 1987 Desjardins et al., 2004 Desjardins et al., 2005 Walsh etal., 1987 * The study denoted with an asterisk is proposed for use by EPA/OPPTfor risk assessment. Shaded studies will not be used to evaluate the risk of HCBD because inadequate test methods or incomplete information were provided for the study. D-2-2 Aquatic Invertebrate Toxicity The toxicity to aquatic invertebrates is summarized below and presented in Table_Apx D-3. The study denoted with an asterisk is proposed for use by EPA/OPPT for risk assessment. Those studies not considered adequate for risk assessment by EPA/OPPT are shaded in the table. Water fleas, (Daphnia magna; 10 animals per replicate) were exposed to nominal concentrations of 0.0015, 0.0022, 0.0032, 0.0046 and 0.0068 mg/L HBCD under flow through conditions for 48 hours (Graves and Swigert, 1997a). On day 0, mean measured exposure concentrations were 0.0021, 0.0018, 0.0018, 0.0026 and 0.0031mg/L and on day two, the mean measured exposure concentrations were 0.0024, 0.0017, 0.0023, 0.0015, and 0.0034 mg/L. The reported 48-hr EC5o was >0.0032 mg/L. Jatzek (Jatzek, 1990) reported a lowest-observed-effect-concentration (LOEC) of 10 mg HBCD/L and an ECso of 146.34 mg HBCD/L for daphnia immobility based on nominal test concentrations that greatly exceeded measured water solubility values for HBCD (0.0488, 0.0147, and 0.0021 u.g/L for a-, (3-, and v-HBCD); however, because the appearance of the test solutions was not reported, it is possible that reduction in daphnid swimming was related to the presence of insoluble material at higher concentrations. For chronic toxicity, hazard was determined based on reduced size (length) of surviving young daphnids, resulting in a measured MATC of 0.0042 mg HBCD/L (Drottar and Krueger, 1998). Page 78 of 97 ------- This study used a flow-through test system and reported additional effects, including decreased reproductive rate and decreased mean weight of surviving young at 0.011 mg HBCD/L. Mortality of adult daphnids in HBCD treatment groups was not significantly different from control mortality. Sublethal effects to invertebrates following chronic exposure were found in supporting studies that assessed endpoints beyond those evaluated in guideline studies. In invertebrates, degenerative changes in the gills of clams (Macoma balthica), manifested by the increased frequency of nuclear and nucleolar abnormalities and the occurrence of dead cells, were observed at nominal concentrations of 0.1 mg HBCD/L (50-day LOEC) (Smolarz and Berger, 2009). Table_Apx D-3: Toxicity of HBCD to Aquatic Invertebrates Test Species Fresh/ Salt Water Duration End- point Cone. (mg/L) Test Analysis Effect References Aquatic Invertebrates - Acute Toxicity *Waterflea (Daphnia magna) Water flea (D. magna Straus) Fresh Fresh 48-hour 48-hour ECso ECso >0.0032 146.34 Exceed WS Flow- through, Measured Static, Nominal Immobilization Immobilization Graves and Swigert, 1997a Jatzek, 1990 Aquatic Invertebrates - Chronic Toxicity Clam (Macoma balthica) *Waterflea (Daphnia magna) Marine Fresh 50-day 21-day LOEC LOEC NOEC 0.1 0.0056 mg/L 0.0031 mg/L Static, Nominal Flow- through, Measured degenerative changes in the gills manifested by the increased frequency of nuclear and nucleolar abnormalities and the occurrence of dead cells reduced length of surviving young Smolarz and Berger, 2009 Drottar and Krueger, 1998 * The study denoted with an asterisk is proposed for use by EPA/OPPTfor risk assessment. Shaded studies will not be used to evaluate the risk of HCBD because inadequate test methods or incomplete information were provided for the study. Toxicity studies for sediment organisms are summarized in Table_Apx D-4. The study denoted with an asterisk is proposed for use by EPA/OPPT for risk assessment. Those studies not considered adequate for risk assessment by EPA/OPPT are shaded in the table. Page 79 of 97 ------- In two prolonged sediment toxicity tests (Thomas et al., 2003a, 2003b) with Hyalella azteca exposed to spiked sediment in the presence of 2 or 5% TOC, no effects were seen at the highest concentration tested (1000 mg HBCD/kg dry weight sediment). Results of a non-GLP range- finding test were submitted in conjunction with the definitive tests that showed reduced survival of H. azteca at 500 mg HBCD/kg dry weight sediment in the presence of 2 or 5% TOC. In another study, Lumbriculus variegatus were tested at nominal test concentrations of 0.05, 0.5, 5, 50, and 500 mg HBCD/kg dry weight sediment. Corresponding measured concentrations were ND, 0.2, 3.1, 28.7, 303.2 mg HBCD/kg dry weight. No HBCD was detected in the overlaying water or in the pore water. Study details were excerpted from a secondary source (Oetken et al., 2001). Table_Apx D-4: Toxicity of HBCD to Sediment Organisms Test Species *Amphipod (Hyalella azteca) Amphipod (H. azteca) *Amphipod (H. azteca) Chironomid (Chironomus riparius) Lumbriculus variegatus Fresh/ Salt Water Fresh Fresh Fresh Fresh Fresh Duration 28-day 28-day 28-day 28-day 28-day End- point LOEC NOEC NOEC NOEC No Dose LOEC NOEC Cone. 500 mg/kg dwt sediment 100 mg/kg dwt sediment 1,000 mg/kg dwt sediment 1,000 mg/kg dwt sediment Not dose- responsive 28.7 mg/kg dwt sediment 3.1 mg/kg dwt sediment Test Analysis Flow- through, Measured Flow- through, Measured Flow- through, Measured N/A Static, Measured Effect reduced survivability Unspecified Unspecified Not dose- responsive reduction in worm number References Thomas et al., 2003a, b Thomas et al., 2003b Thomas et al., 2003a Thomas et al., 2003a Oetken et al., 2001 * The study denoted with an asterisk is proposed for use by EPA/OPPTfor risk assessment; N/A = not applicable Shaded studies will not be used to evaluate the risk of HCBD because inadequate test methods or incomplete information were provided for the study. D-2-3 Fish Toxicity The toxicity to fish is summarized below and presented in Table_Apx D-5. The study denoted with an asterisk is proposed for use by EPA/OPPT for risk assessment. Those studies not considered adequate for risk assessment by EPA/OPPT are shaded in the table. Page 80 of 97 ------- Acute Effects Rainbow trout (Oncorhynchus mykiss) were exposed to mean measured concentration of HBCDs of 0.00075, 0.0015, 0.0023, 00.23 and 0025 mg/L (with solvent) under flow through conditions for 96 hours. No effects were observed at any test concentration (Graves and Swigert, 1997b). Bluegill sunfish (Lepomis macrochirus) were exposed to nominal concentrations ranged from 10 to 100 mg/L under static conditions for 96 hours. These test concentrations exceeded the test substance water solubility. A white flocculate was formed on the surface of the water in all test solutions. No effects were seen at the highest test concentration of 100 mg/L (Calmbacher, 1978)(Calmbacher, 1978). Acute effects observed following acute exposure were found in a 96-hour test with zebrafish embryos (Deng et al., 2009). In a 96-hour toxicity study with zebrafish embryos, increased malformation rate was observed at a nominal concentration of 0.1 mg HBCD/L and decreased survival and increased heart rate were observed at a nominal concentration of 0.05 mg HBCD/L (lowest concentration tested); an ECso or LCso value was not reported (Deng et al., 2009). Chronic Effects A chronic study in rainbow trout conducted following EPA recommended guidelines, found no effects at concentrations approaching saturation (Drottar et al., 2001). No effects were observed in European flounder (Platichthys flesus) following 78 days of diet or sediment exposure to maximum concentrations of 3000 u.g HBCD/g lipid in food and 8000 u.g HBCD/g total organic carbon (TOC), respectively (Kuiper et al., 2007). Sublethal effects to fish following chronic exposure were found in supporting studies that assessed endpoints beyond those evaluated in guideline studies. Effects observed in fish include increased formation of reactive oxygen species (ROS) resulting in oxidative damage to lipids, proteins, and DNA, decreased antioxidant capacities in fish tissue (e.g., brains, hepatocytes, or erythrocytes), and increasing levels of ethyoxyresorufin-O-deethylase (EROD, detoxification enzyme) and pentoxyresorufin-O-deethylase (PROD, detoxification enzyme) levels in hepatocytes of fish exposed to the nominal concentration of >0.1 mg HBCD/L (corresponds to ~0.2 mg HBCD/g whole fish [wet weight]) for 42 days (Zhang et al., 2008). Indications of endocrine disruption were reported following dietary exposure to HBCD that impacted the thyroid system of juvenile rainbow trout (Oncorhynchus mykiss). Each of the diastereoisomers of HBCD (administered separately via diet at concentrations of 5 ng/g of a-, P-, or y-HBCD) disrupted thyroid homeostasis, as indicated by lower free circulating triiodothyronine (T3) and thyroxine (T4) levels (Palace et al., 2010; Palace et al., 2008). Page 81 of 97 ------- Table_Apx D-5: Toxicity of HBCD to Fish Test Species Fresh/ Salt Water Duration End- point Cone. (mg/L) Test Analysis Effect References Fish - Acute Toxicity Bluegill sunfish (Lepomis macrochirus) *Rainbow trout (Oncorhynchus mykiss) Zebra fish embryos (Danio rand] Fresh Fresh Fresh 96-hour 96-hour 96-hour LCso LCso LCso >100 >0.0025 0.05 Static, Nominal Flow- through, Measured Static, Nominal Mortality Mortality decreased survival, reduced heart rate Calmbacher, 1978 Graves and Swigert, 1997b Denget al., 2009 Fish - Chronic Toxicity Rainbow trout (Oncorhynchus mykiss) Rainbow trout (Oncorhynchus mykiss) Chinese rare minnow (Gobiocypris rarus) European flounder (Platichthysflesus) *Rainbow trout (O. mykiss) Fresh Fresh Fresh Fresh 88-day 28-day 42-day 78-day 32-day NOEC LOEC MATC NOEC LOEC 0.0037 0.1 0.032 8,000 ng/g TOC 3000 ng/g lipid in muscle 5 ng/g of a-, P-, or v- HBCD Flow- through, Measured Intraperi- toneal injection using 50 and<500 mg/kg-bw Static, Nominal Sediment exposed Diet exposed Diet exposed No effects Significant catalase activity at 50 and<500 mg/kg-bw; EROD activity; LSI increase; No effects on blood plasma, vitellogenin levels or DNA adducts formation. DNA damage in erythrocytes; induction of EROD and PROD in hepatocytes; ROS formation in brain tissue No effects on behavior, survival, growth rate, relative liver and gonad weights thyroid effects Drottar et al., 2001 (Ronisz et al., 2004) Zhang et al., 2008 Kuiper et al., 2007 Palace et al., 2010 * The study denoted with an asterisk is proposed for use by EPA/OPPTfor risk assessment. Shaded studies will not be used to evaluate the risk of HCBD because inadequate test methods or incomplete information were provided for the study. Page 82 of 97 ------- D-3 Toxicity to Terrestrial Organisms D-3-1 Terrestrial Plant Toxicity Available toxicity studies for terrestrial plants are summarized in Table_Apx D-6. Studies using monocot and dicot plant species exhibited no toxicity up to the maximum test concentration of 5,000 mg HBCD/kg soil (Porch et al., 2002). Table_Apx D-6: Toxicity of HBCD to Terrestrial Plants Test Species Corn (Zea mays) Cucumber (Cucumis sativa) Onion (Allium cepa) Ryegrass (Lolium perenne) Duration 21-day End-point NOEC Cone. >5,000 mg HBCD/kg soil Test Analysis Nominal Effect No treatment- related effects on emergence, survival or growth References Porch et al., 2002 D-3-2 Soil Invertebrate Toxicity The available toxicity study in soil organisms summarized below and presented in Table_Apx D-7 is adequate for risk assessment*. Aufderheide et al., (Aufderheide et al., 2003) reported an ECioand no-observed-effect- concentration (NOEC) values of 21.6 and 128 mg HBCD/kg dry soil, respectively, based on reproductive effects in earthworms following 56 days of exposure. High variability in the data at the lower test concentrations (as indicated in ECB, 2008) resulted in wide confidence limits for the ECio, differences from the control that were not significant, and a NOEC that was greater than the ECio. Two worms from the lowest test concentrations were lost during the study and treated as dead (as reported in ECB, 2008), which may have contributed to the high variability by reducing the sample size at the lowest test concentration; however, study details, data tables, and statistical methodology were not available. Table_Apx D-7: Toxicity of HBCD to Terrestrial Invertebrates Test Species *Earthworm (Eisenia fetida) Duration 56-day End-point ECio NOEC Cone. 21.6 mg HBCD/kg dry soil 128 mg HBCD/kg dry soil Test Analysis Nominal, Static, Soil Effect reproduction References Aufderheide et al., 2003 Page 83 of 97 ------- D-3-3 Avian Toxicity Available toxicity studies to avian species are summarized in Table_Apx D-8. Japanese quail eggs exposed for 6 weeks to an isomeric mixture of HBCD in the diet experienced a reduction in hatchability at all tested concentrations (12-1000 ppm) (MOEJ, 2009). Additional effects included a significant reduction in egg shell thickness starting at 125 ppm, decreases in egg weights and egg production rates starting at 500 ppm, increases in cracked eggs starting at 500 ppm, and adult mortality at 1000 ppm. A subsequent test, conducted at lower dietary concentrations, determined a lowest-observed-adverse-effect-level (LOAEL) and no-observed-adverse-effect-level (NOAEL) values of 15 and 5 ppm, respectively, based on significant reduction of survival of chicks hatched from eggs of HBCD-fed quails (MOEJ, 2009). In another study, a number of effects were reported in American Kestrels exposed in ovo to 164.13 ng HBCD/g wet weight (Kobiliris, 2010). Table_Apx D-8: Toxicity of HBCD to Avian Species Test Species Japanese quail (Coturnix coturnix japonica) American kestrel (Falco sparverius) Duration 6-week End-point LOAEL LOAEL Cone. 125 ppb 15 ppm (2.1 mg/kg body wt/day 5 ppm 164.3 ng/g wet weight of egg Test Analysis Diet exposed In ovo exposed Effect reduction in hatchability reduced chick survival reduced corticosterone response in male nestling kestrels, reduced flying activities in juvenile males, delayed response time to predator avoidance in juvenile females References MOEJ, 2009 Kobiliris, 2010 Page 84 of 97 ------- Appendix E Human Health Hazard Study Summaries E-l Toxicokinetics For humans, there is a potential for oral, inhalation and dermal exposure. Available toxicokinetics data in rodents indicate that HBCD is moderately absorbed via the gastrointestinal tract, metabolized, and distributed to a number of nonfat tissues including blood, muscle, and the liver, where it accumulates unchanged (Arita et al., 1983; Brandsma et al., 2009; Hakk et al., 2012; Reistad et al., 2006; Sanders et al., 2013; Szabo et al., 2010; Szabo et al., 2011b; van der Ven et al., 2009; van der Ven et al., 2006; Yu and Atallah, 1980). Elimination of HBCD is predominantly via feces (as the parent compound), but it is also eliminated in urine (as secondary metabolites) (Arita et al., 1983; Yu and Atallah, 1980; Szabo et al., 2010). In humans, HBCD has been detected in breast milk, adipose tissue, blood, and both maternal and umbilical serum (Abdallah and Harrad, 2011; Antignac et al., 2008; Covaci et al., 2006; Fangstrom et al., 2008; Fangstrom et al., 2005; Johnson-Restrepo et al., 2008; Kakimoto et al., 2008; Meijer et al., 2008; Rawn et al., 2014; Thomsen et al., 2007; Weiss et al., 2006) E-2 Acute Toxicity Studies Several acute toxicity studies in rats and rabbits by the oral, dermal, and inhalation routes with HBCD are available (BASF, 1990; Gulf South Research Institute, 1988; IRDC, 1977,1978a, 1978b; Lewis and Palanker, 1978; Momma et al., 1993). The acute toxicity of HBCD is low via the oral route in rats and low via the dermal route in rabbits (Lewis and Palanker, 1978), with LDso values >680 mg/kg-bw. Acute inhalation exposure to HBCD resulted in some minor symptoms (such as eye squint, slight dyspnea, salivation, lacrimation, and nasal discharge), but no LCso has been identified. The acute toxicity of HBCD is summarized in Table_Apx E-l (oral) and Table_Apx E-2 (inhalation). Table_Apx E-l: Acute Oral Toxicity of HBCD Species/strain/ test Rat/Charles River/LDso Rat/Charles River CD/ LD5o test of HBCD residue Exposure Single gavage (corn oil) Single gavage (corn oil) Result LD5o > 10,000 mg/kg LD5o = 1258 mg/kg (male); 680 mg/kg (female) Notes No mortality; transient hypoactivity and diarrhea; corneal opacity and ptosis in 3/5 males, which did not resolve by end of 14-day observation. Tested Firemaster 100; increased activity, eye squint, dyspnea, lacrimation, and nasal discharge. Reference IRDC, 1977 IRDC, 1978a Page 85 of 97 ------- Species/strain/ test Mouse/NR/LDso Rat/Sprague- Dawley/limit test of HBCD bottoms Rat/NR/LDsotest Exposure Single gavage (30% aqueous tragacanth) Single gavage (corn oil) Single gavage (corn oil) Result LD5o >6,400 mg/kg; data not shown LD5o >5,000 mg/kg; clinical signs LD5o > 10,000 mg/kg Notes Weight loss in all animals; recovery noted by conclusion of study. 7-day observation period; increasing apathy, trembling and late mortalities; peritonitis at necropsy. This study tested HBCD bottoms described as black solids, lacrimation, and facial swelling resolved by post-exposure day 4; no gross lesions were observed. No significant changes observed. Reference BASF, 1990 Pharmakon Research International Inc., 1990 Gulf South Research Institute, 1988 Abbreviations: NR, not reported Table_Apx E-2: Acute Inhalation Toxicity of HBCD Species/strain/test Rat/Charles River/LD5o Rat/Wistar/limittest Rat/Charles River/LC5o test of Firemaster 100 residue (liquid) Rat/NR/LCsotest Exposure 4-hr exposure to Firemaster 100 dust: 202. 14 mg/L 1-hr exposure to 200 mg/L GLS-S6- 41A (highest possible concentration) 4-hr exposure to Firemaster 100 residue: 22.9 mg/L 1 hr. whole-body inhalation exposure to GLS-S6-41A Result No mortality; slight dyspnea at the end of exposure only No mortality; no clinical signs Notes All rats gained weight over the observation period, but no control data were reported. All rats gained weight over the observation period, but no control data were reported. One death (female, Day 3 post-exposure); dyspnea, salivation, lacrimation, and nasal discharge during exposure, resolved by Day 3; significant body weight loss through Day 3, but beginning to recover by Day 14 post exposure; signs of nasal and lung irritation at gross necropsy of rat that died. LC5o >200 mg/L No significant effects observed. Reference IRDC, 1977 Lewis and Palanker, 1978 IRDC, 1978b Gulf South Research Institute, 1988 Abbreviations: NR, not reported E-3 Repeated-Dose Toxicity Studies Short-term and subchronic toxicity studies on HBCD are available and summarized Table_Apx E-3. In these studies, HBCD demonstrated effects on the thyroid and liver (Chengelis, 2002; Page 86 of 97 ------- Chengelis, 1997, 2001; van der Ven et al., 2006). However, most of these effects were not significant after a recovery period of 14 days and 28 days, respectively (Chengelis, 2002; Chengelis, 1997, 2001). Older short-term and chronic toxicity studies using an HBCD product that is no longer manufactured concluded that observed liver findings were adaptive rather than adverse (Zeller and Kirsch, 1969,1970). Developmental behavioral defects were observed in 3-month old mice after a single oral exposure on postnatal day (PND) 10 (Eriksson et al., 2006). This following summary is extracted from the hazard characterization found in the supporting documents for Initial Risk-Based Prioritization of High Production Volume Chemicals (EPA, 2008a): The potential toxicity from repeated oral exposure to HBCD was assessed in a variety of studies in laboratory animals. Liver effects were observed in several studies but based on the inconsistency of effects between studies and sexes, and lack of dose-response, it is not clear if the observed effects are treatment-related. Effects on the thyroid (one or both sexes) were observed at moderate to high doses in some repeated-dose studies but not others, but could be due to the fact that the thyroid system was not thoroughly studied in the early studies. More recent studies showed increased thyroid weights in females only. One study indicates decreased serum T4 and increased serum TSH in both sexes, whereas another study only shows effects in females. Taken together, however, the data are suggestive of possible treatment- related thyroid effects in adult animals. Several recent in vivo and in vitro studies have been conducted to try and elucidate the possible mechanisms for both the observed liver and thyroid effects, but with no clear conclusions. Functional observation battery and motor activity evaluations in adult animals showed no evidence of neurotoxicity. Table_Apx E-3: Repeated-Dose Toxicity of HBCD Species/strain Sprague-Dawley rats Sprague-Dawley rats Exposure 28-days 90-days Result Liver weight increase from the lowest dose (940 mg/kg-day) in both sexes. Thyroid hyperplasia from the lowest dose (940 mg/kg-day) in both sexes. Liver weight increases from the lowest dose (120 mg/kg-day) in both sexes. No histopathological effects in thyroid were reported. Notes The authors attributed the increased liver weight to hyperactivity as a result of increased thyroid activity and concluded the increased liver weights were not pathologic. NOAEL = 940 mg/kg-day. Demonstrates a low order of toxicity and may reflect a reversible adaptive change. Data supports that the liver and thyroid glands are targets. Reference Zeller and Kirsch, 1969 Zeller and Kirsch, 1970 Page 87 of 97 ------- Species/strain Sprague-Dawley rats Sprague-Dawley rats Wistar rats Exposure 28-days 90 days 28 days Result Liver weight increase in females from the lowest dose (125 mg/kg- day) and in males from the mid dose (350 mg/kg-day). No histological effects observed in the thyroid in either sex. Liver weight increase from the lowest dose (100 mg/kg-day) in both sexes. Liver weight increases in females at 23 mg/kg-day Notes The effects on the liver especially in female rats indicate a LOAELof 125 mg/kg-day. Thyroid weight was increased from the mid- dose in females (300 mg/kg-day), but not in males. Serum T4 was decreased and TSH increased in all dose groups of both sexes. LOAEL of 100 mg/kg-day based on increases in liver weights and changes in thyroid serum concentrations. Most sensitive endpoint was a 10% increase in thyroid weight in females at 3 mg/kg-day. Reference Chengelis, 1997 Chengelis, 2001 Van der Ven et al., 2006 E-4 Reproductive and Developmental Toxicity Studies The available toxicity data for HBCD also includes evidence of reproductive, developmental, and neurological toxicity in rats and mice which are summarized in Table_Apx E-4 and Table_Apx E-5. An exposure-response array for reproductive and developmental toxicity is presented in Figure_Apx 2.6.4-2 and for neurological effects is presented in Figure_Apx 2.6.4-3. Information on the developmental and reproductive toxicity of HBCD comes from a single- generation reproductive toxicity study (van der Ven et al., 2009), a study incorporating gestational and post-natal exposure (Saegusa et al., 2009), a two-generation reproduction toxicity study (Ema et al., 2008), a gestational exposure study (Murai et al., 1985), and a neurotoxicity study of adult mice neonatally exposed to HBCD (Eriksson et al., 2006). Page 88 of 97 ------- Table_Apx E-4: Reproductive and Developmental Toxicity of HBCD* Species/strain Wistar rats CrhCD(SD) rats Wistar rats Crj:CD(SD)IGS rats Wistar rats Exposure One full spermatogenic or two full estrous cycles (males: 70 d prior to mating; females: 14 d prior to mating) and continued during pregnancy and lactation for a total of 11 wks post weaning 10 wks prior to mating and through gestation, lactation, and for two generations (multi- generation reproductive toxicity study) GD 0-20 GD 10-PND 20 (weaning) See Van der Ven et al., 2009 Result No exposure related changes in reproductive parameters, including mating success, time to gestation, gestation duration, number of implantation sites, litter size, and sex ratio. A significant decrease in the number of primoridal follicles in the ovary; decreased size of the thyroid follicles in both sexes and an increase in pup mortality during lactation No significant changes in the number of implants, resorptions, live or dead fetuses or external, visceral or skeletal anomalies No exposure-related changes in reproductive parameters; however, increased thyroid weight and decreased serum Ta in male offspring at ~146 mg/kg-day Effect on Brainstem Auditory Evoked Potentials (BAEPs) observed in the low frequency range and only in male off- spring (see text) Notes NOAEL ~100 mg/kg-day (highest dose tested) LOAEL- 101 mg/kg-day NOAEL- 10 mg/kg-day NOAEL- 750 mg/kg-day (highest dose tested) NOAEL -1505 mg/kg-day (highest dose tested) Neurobehavioral assessment of the rats in the Van der Ven et al., 2009 study. Reference Van der Ven et al., 2009 Ema et al., 2008 Murai et al., 1985 Saegusa et al., 2009 Lilienthal et al., 2009 "Only studies considered adequate for risk assessment are presented in the table. Page 89 of 97 ------- Signs of developmental toxic effects in Wistar rats included immune system effects, indications of liver toxicity, and decreases in bone mineral density at very low doses, i.e., <1.3 mg/kg-day (van der Ven et al., 2009); however, the authors noted that the vehicle used (corn oil) may have affected observations at higher doses, including: increased mortality during lactation, decreased liver weight in males, decreased adrenal weight in females, decreased plasma cholesterol in females, and other immunological markers of toxicity. Saegusa et al. (2009) observed an increased relative thyroid weight and decreased triiodothyronine (T3) levels in Fl male Sprague-Dawley rats at postnatal week (PNW) 11 following dietary exposure to 1,000 ppm (approximately 146.3 mg/kg-day) HBCD. Saegusa et al. (2009) also reported a significant reduction in the number of CNPase-positive oligodendrocytes at 10,000 ppm (approximately 1,504.8 mg/kg-day). Developmental toxicity was not observed in Fl Wistar rats following dietary exposure to HBCD during gestation (Murai et al., 1985). Ema et al. (2008) reported a reduced viability index on Day 4 and Day 21 of lactation among second generation (F2) offspring at 15,000 ppm (approximately 1,363 mg/kg-day). Ema et al. (2008) observed additional developmental effects at doses as low as 1,500 ppm (approximately 115 and 138 mg/kg-day for Fl males and females, respectively), including: an increase in dihydrotestosterone (DHT) in Fl males and an increased incidence of animals with decreased thyroid follicle size in both sexes and generations. These authors reported no effects on sexual development indicated by anogenital distance, vaginal opening, or preputial separation among Fl or F2 generations. Reproductive toxic effects have also been observed in Ema et al. (2008). A decrease in the number of primordial follicles in first generation (Fl) female Crl:CD(SD) rats at 1,500 ppm (approximately 138 mg/kg-day) and a significant increase in the number of litters lost in the Fl generation at 15,000 ppm (approximately 1,363 mg/kg-day) were reported. No other treatment-related adverse effects were reported in any generation for indicators of reproductive health, including: estrous cyclicity, sperm count and morphology, copulation index, fertility index, gestation index, delivery index, gestation length, number of pups delivered, number of litters, or sex ratios. Neither van der Ven et al. (2009) nor Saegusa et al. (2009) observed reproductive effects at the doses tested (up to 100 mg/kg-day in Wistar rats and approximately 1,504.8 mg/kg-day in Sprague-Dawley rats, respectively). No standard neurotoxicity or developmental neurotoxicity studies on HBCD are available. Information on neurotoxicity was obtained from Functional Observational Battery, locomotor activity evaluations, neurobehavioral testing, surface righting reflex, negative geotaxis reflex, mid-air righting reflex, and brainstem auditory evoked potentials (BAEPs) in several repeated- dose and reproductive toxicity studies. For example, in a subacute toxicity study, HBCD was administered orally by gavage in corn oil to Sprague-Dawley CrhCD BR rats for 28 days at doses of 0, 125, 350, or 1,000 mg/kg-day (6 Page 90 of 97 ------- rats/sex/dose in 125 and 350 mg/kg-day groups and 12 rats/sex/dose in the control and 1,000 mg/kg-day groups) (Chengelis, 1997). At the end of 28 days, 6 rats/sex/dose were necropsied, while the remaining rats in the control and 1,000 mg/kg-day groups were untreated for a 14- day recovery period prior to necropsy. Functional Observational Battery and motor activity evaluations were carried out prior to study initiation, during the last week of HBCD administration (Week 3), and during the recovery period (Week 5). No changes in the Functional Observational Battery and motor activity tests were reported. In another subchronic toxicity study, (Chengelis, 2002; Chengelis, 2001) administered HBCD by oral gavage in corn oil daily to Crl:CD(SD)IGS BR rats (15/sex/dose) at dose levels of 0,100, 300, or 1,000 mg/kg-day for 90 days. At the end of 90 days, 10 rats/sex/dose were necropsied, while the remaining rats were untreated for a 28-day recovery period prior to necropsy. Functional Observational Battery and locomotor activity evaluations were carried out on 5 a ni ma Is/sex/dose prior to study initiation, during the last week of HBCD administration (Week 13), and during the recovery period. There were no treatment-related effects on Functional Observational Battery and locomotor activity observed. In a one-generation study that included additional immunological, endocrine and neurodevelopmental endpoints, van der Ven (2009) exposed Wistar rats (10/sex/dose) to a composite mixture of technical-grade HBCD in the diet at concentrations resulting in doses of 0.1, 0.3, 1.0, 3.0, 10, 30, or 100 mg/kg-day. This study followed OECD Guidelines 415 and 407 (OECD, 1983,1995). Just prior to the end of the study, at approximately 8 weeks of age, the remaining Fl pups were assigned to separate groups for necropsy (5/sex/dose), immunological testing (4 males/dose), or neurobehavioral testing (6/sex/dose plus additional males and females from selected dose groups). Lilienthal et al. (Lilienthal et al., 2009) reported the results of the neurobehavioral assessment from the one-generation study conducted by van der Ven et al. (2009). Lilienthal et al. (2009) examined 110 day-old Fl rats for haloperidol-induced catalepsy to determine possible effects of HBCD on the dopaminergic system. One month after the catalepsy measurements, brainstem auditory evoked potentials (BAEPs) to broadband click and frequency-specific tone stimuli were recorded in males and females to examine potential effects on auditory functions. The study authors concluded that the observed dose-dependent decreases in latencies may be due to HBCD-related effects on dopaminergic activity or to HBCD- related induction of metabolizing liver enzymes resulting in enhanced metabolism of haloperidol. HBCD-related effects on BAEP were observed in the low frequency range and only in male offspring. The study authors hypothesized that HBCD exerts a cochlear effect on males based on the results of the BAEPs. The authors noted that an alternative explanation could be HBCD-induced changes in retinoids, which may be involved in development of the inner ear. Too little information is available in this study to determine the significance of its findings. Finally, in a two-generation reproductive toxicity study, Ema et al. (2008) administered HBCD to Crl:CD(SD) rats conducted according to OECD Guideline 416 (OECD, 2001), US EPA Guidelines (EPA, 1991,1996), and good laboratory practices (GLP). Groups of male and female rats Page 91 of 97 ------- (24/sex/dose) were fed HBCD (as a mixture of a-HBCD, (3 -HBCD, and v-HBCD with proportions of 8.5, 7.9, and 83.7%, respectively) in the diet at concentrations of 0,150,1,500, or 15,000 ppm from 10 weeks prior to mating through mating, gestation, and lactation. Reproductive and developmental milestones were monitored, including: surface righting reflex, negative geotaxis reflex, mid-air righting reflex, and anogenital distance. F2 females exposed to 15,000 ppm HBCD completed the mid-air righting reflex (76.9%) than control F2 females (100%). These findings were not consistent over generations or sexes and were not considered treatment related. No other effects of HBCD exposure on the development of reflexes were observed in either Fl or F2 progeny. An additional study on the neurotoxicity of HBCD is available. Eriksson et al. (2006) observed effects on spontaneous motor behavior, learning, and memory in adult NMRI mice following exposure to HBCD on postnatal day (PND) 10. At 0.9 mg/kg, the authors reported significantly reduced mean locomotor activity. However, this study was not conducted according to current guidelines (EPA, 1998b) and GLP, therefore EPA reserves judgment on the significance of these findings. The authors used too few dose groups and the behavioral alterations were induced at doses that did not produce clinical signs or affect weight gain. Additionally, effects due to litter size were not considered. However, this study did demonstrate good repeatability for control values and for relevant active substances tested several times. an en _ Art _ n , t 138 i ^ 146.3 .. T21.3 14.3 "114 (Male) | Dose Range ALnAEL • NOAEL • BMDL A 13.5 I 0.9 _ o.056 Decreased number Increased pup Increasedthvroid Effects not observed Decreased Decreased bone of primordial follicles mortality in F2 weightand -rat [Muraietal., habituation, mineral density in in Fl females-rat generation-rat(Ema decreasedTSinFl 1985) locomotion, and females-ratfvan der [Emaetsl., 2008) etal.. 2008) males - rat [Saegusa rearing-mouse Venetal., 2009) etal., 2009) (Eriksson etal., 2006) Figure_Apx 2.6.4-2: Exposure-Response Array for Developmental and Reproductive Toxicity Studies of HBCD Page 92 of 97 ------- Table_Apx E-5: Summary of Effects in Parental and Fl Rats After Dietary, Gestational, Lactational and Postnatal Exposure to HBCD Generation HBCD (ppm in diet) Males' 0 100 1,000 10,000 Females0 0 100 1,000 10,000 FO Thyroid Relative weight (mg/100 gBW) Histopathology: diffuse follicular cell hypertrophy (±/+/++/+++)a ND ND ND ND ND ND ND ND 5.73 3/10 (0/3/0/0)b 6.75 5/10 (2/3/0/0) 6.30 6/10 (1/3/2/0) 7.47* 9/10# (0/3/4/2)§§ Fl Relative organ weights, PND 20 Liver (g/100 g BW) 3.68 3.82 3.98 4.66* 3.77 3.83 4.01 4.83* Relative organ weights, PNW 11 Liver (g/100 g BW) Thyroid (mg/100 g BW) Epididymides (mg/100 g BW) 3.45 4.85 0.23 3.81" 5.66 0.21* 3.58 5.78* 0.22 3.53 6.20" 0.21 3.35 8.20 NA 3.59 6.84 NA 3.44 7.35 NA 3.30 7.72 NA Thyroid-related hormones PND 20 T3 (ng/ml) T4(ug/dl) TSH (ng/ml) 1.09 4.39 5.40 1.13 4.20 6.66 1.06 4.78 6.07 0.93" 4.20 7.00* ND ND ND ND ND ND ND ND ND ND ND ND PNW 11 T3 (ng/ml) T4(ug/dl) TSH (ng/ml) 0.96 4.77 4.74 0.93 4.84 5.81 0.88* 5.21 5.36 0.89" 5.20 4.96 ND ND ND ND ND ND ND ND ND ND ND ND Histopathology PND 20 Liver: Vacuolar degeneration, liver cells, diffuse (+/++)a 0/10 0/10 0/10 6/10* (6/0) 0/10 0/10 0/10 6/10* (0/6)§§ PNW 11 Adrenal: Vacuolar degeneration, diffuse, cortical cells (+/++)a 0/10 0/10 0/10 4/10* (2/2)§ ND ND ND ND Abbreviations: BW, body weight; PND, postnatal day; ppm, parts per million; PNW, postnatal week; ND, no data; NA, not applicable; aGrade of change: (±) minimal; (+) slight; (++) moderate; (+++) severe; bNumber of animals with each grade; cn=10 rats/sex/group; * Significantly different from the controls by Dunnett's test or Dunnett-type rank-sum test (p<0.05) " Significantly different from the controls by Dunnett's test or Dunnett-type rank-sum test (p<0.01) "Significantly different from the controls by Fisher's exact probability test (p<0.05) § Significantly different from the controls by Mann-Whitney's U-test (p<0.05) §§ Significantly different from the controls by Mann-Whitney's U-test (p<0.01) Source: (Saegusa et al., 2009) Page 93 of 97 ------- •? 1 ! * » 0 o (1 , , ,, o Ik Q— — f— — i— — i— — i— — «s— _| * O TO ^^r >• x^x^x — '> 5=-^ 5^-^ ^r^ Q C LO g 1 1 g | 1 i | SI | | | | | I | g | § j j 1 — r « ° ^ "s ^ "- ^ ^ 1 ° 1 °- — ^ S •- | 1 || Jl | | 3,1 SE .is II" ^ i W1 Ncurobchavior 1 1 c c . Sec physio Cha ) 3 o i 5 tro- ogical TgCS FOB evaluations consists of open field, home cage, sensory, neut omuscular and physiological observations Figure_Apx 2.6.4-3: Exposure-Response Array for Neurological Effects of HBCD Source: (EPA, 2014d) E-5 Skin Irritation and Sensitization Studies The available literature indicates that HBCD is not a dermal irritant in guinea pigs at concentrations up to 0.5 ml, but one study found HBCD to be a mild skin allergen (Momma et al., 1993). Acute eye irritation studies in rabbits showed HBCD to be a mild transient ocular irritant (Gulf South Research Institute, 1988). E-6 Genotoxicity and Cancer Studies A limited number of studies investigated the genotoxicity of HBCD. These studies, summarized in Table_Apx E-6, indicate that HBCD is not likely to be genotoxic. The majority of these studies were standard Ames tests for detecting mutagenic potential in Salmonella typhimurium (S. typhimurium). Most Ames tests conducted with HBCD yielded negative results (Huntington Research Center, 1978; IBT, 1990; Litton Bionetics Inc, 1990; Pharmakoligisches Institute, 1978; SRI, 1990; Zeiger et al., 1987). Two Ames tests showed positive, dose-dependent results for strain TA1535; one for TA1535 only using a liquid residue of HBCD in DMSO (IBT, 1990); and one for strains TA1535 and TA100 using an unidentified Page 94 of 97 ------- mixture characterized only as HBCD bottoms in acetone (Ethyl Corporation, 1985a). Both of these strains detect reversions by base pair substitution. However, the Ames tests in the strains that were positive in these two studies (TA1535 and TA100) were negative in the other studies cited above. In mammalian systems, a reverse mutation assay with Chinese hamster ovary (CHO) Sp5 and SPD8 cell lines exposed to HBCD yielded positive results (Helleday et al., 1990). A test of unscheduled DNA synthesis with rat hepatocytes exposed to HBCD bottoms was also positive with a dose-response relationship (Ethyl Corporation, 1985b). The reverse mutation assay in CHO cells (Helleday et al., 1999) and the unscheduled DNA synthesis assay in F344 hepatocytes (Ethyl Corporation, 1985b) were not repeated by any other group. Several assays performed to determine the genotoxicity of HBCD were negative even when testing at cytotoxic concentrations, including: one in yeast (Litton Bionetics Inc., 1990), one detecting chromosomal aberrations in human peripheral lymphocytes in vitro (Microbiological Associates Inc, 1996), and one in vivo mouse micronucleus test following intraperitoneal injection of HBCD (BASF, 2000). Several previous assessments have concluded that based on the lack of mutagenicity in vitro and in vivo, HBCD does not have genotoxic potential in vitro or in vivo (EC, 2008; Environment CA and Health CA, 2011; NICNAS, 2012; OECD, 2007). EPA agrees with this conclusion. Existing assessments have also concluded, based on genotoxicity information and one limited lifetime study, that HBCD is not carcinogenic (NICNAS, 2012; TemaNord, 2008) or that further study of carcinogenicity is not warranted (EC, 2008; OECD, 2007). However, the only available dietary study evaluating the carcinogenic potential of HBCD in mice is not considered adequate to draw conclusions regarding carcinogenicity (EC, 2008; Environment CA and Health CA, 2011; EPA, 2014b; OECD, 2007). Given this data gap, EPA's HBCD assessment will not include carcinogenicity assessment. Page 95 of 97 ------- Table_Apx E-6: Genotoxicity of HBCD Test/species/strain/ route Test doses (per plate)3 Results'3 -S9 +S9 Notes Reference Eukaryotic systems, in vitro S. typhimurium TA98, TA100, TA1537 S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 S. typhimurium TA98, TA100, TA1535 S. typhimurium TA98, TA100, TA1535, TA1537 S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 S. typhimurium TA98, TA100, TA1535, TA1537 S. typhimurium TA98, TA100, TA1535, TA1537, TA1538 3,000 ng in DMSO 250 ug (Firemaster, FM- 100, Lot 53, white powder) in DMSO 1,000 ug (FM- 100, Lot 3322, liquid residue) in DMSO 10,000 ug in DMSO 10,000 ug in DMSO 5,000 ug in DMSO 50 Ug (HBCD bottoms ) in acetone 50 ug - - - - - - + (TA1535 and 100 only) - - - + (TA1535 only) - - - + (TA100 only) - Doses >1,000 u.g were partially insoluble. Doses >250 u.g were insoluble. Positive in TA1535 at highest dose only; lower doses showed positive trend with dose. Insoluble at 10,000 ug. No cytotoxicity observed. Dose- response only in TA1535 (-S9) >100 ng/plate. TA100 positive at highest dose only (5,000 ug/plate). All doses had a black precipitate thought to be carbon. Pharmako- logisches Institute, 1978 IBT, 1990 Huntingdon Research Center, 1978 Zieger et al., 1987 SRI, 1990 Ethyl Corporation, 1985a Litton Bionetics, 1990 Prokaryotic non-mammalian systems, in vitro Saccharomyces cerevisiae D4 50 ug - - Litton Bionetics, 1990 Mammalian systems, in vitro Chromosomal aberration test In human peripheral blood lymphocytes 750 ng/mL (-S9) and 250 ng/mL (+S9) in DMSO -m -m Doses 750 - 2,500 were partially insoluble, and fully insoluble >2,500 ng/mL Repeated test for two harvest time points: 20 hr (- S9) or 4 hr (+S9) incubations, and 20 or 44 hr incubations (-S9 and +S9). Microbiological Associates, 1996 Page 96 of 97 ------- Test/species/strain/ route Unscheduled DNA Synthesis rat/F344 male/primary hepatocytes Reversion assay CHO/V79/Sp5 and SPD8 Intragenic recombination at hprt locus in Sp5 (non- homologous recombination) and SPD8 (homologous recombination) duplication cell lines Test doses (per plate)3 5-1,000 ug/well in acetone (HBCD bottoms) 3-20 ng/mL in DMSO Results'3 -S9 + + +S9 NA NA Notes Five highest doses (from 5 Hg/well) showed an increased response with dose over solvent control, but only four highest were statistically significant (x2). Highest dose (1,000 ng/well) was cytotoxic. A statistically significant increase in reversion frequency was observed in both assays in the highest dose group as determined by linear regression analysis. Reference Ethyl Corporation, 1985b Helleday et al., 1999 Mammalian systems, in vivo Micronucleus test mouse/NMRI/intraper itoneal injection 2,000 mg/kg in DMSO -m NA BASF, 2000 aLowest effective dose for positive results; highest dose tested b+ = positive, ± = equivocal or weakly positive, - = negative, T = for negative results. cytotoxicity, NA = not applicable, ND = no data. Page 97 of 97 ------- |