# IUCLID
Data Set

Existing Chemical : ID: 25637-99-4 Memo : HBCD CAS No. : 25637-99-4 EINECS Name : hexabromocyclododecane EC No. : 247-l 48-4 TSCA Name : Cyclododecane, hexabromoMolecular Formula : C12H18Br6

Producer related part Company : ALBEMARLE CORPORATION Creation date : 30.11.2004 Substance related part Company : ALBEMARLE CORPORATION Creation date : 30.11.2004
status Memo Printing date : 31.03.2005 Revision date

. . Date of last update : 12.01.2005
Number of pages : 88 Chapter (profile) : Chapter: 1,2, 3, 4, 5, 6, 7, 8, 10 Reliability (profile) : Reliability: without reliability, 1,2, 3, 4 Flags (profile) : Flags: without flag, confidential, non confidential, WGK (DE), TA-Luft (DE), Material Safety Dataset, Risk Assessment, Directive 67/548/EEC, SIDS

1 I88

## ‘l. General Information Id Date 25637-99-4 31.03.2005

1.0.1 APPLICANT AND COMPANY INFORMATlON

Type Name Contact person Date Street Town Country Phone Telefax Telex Cedex Email Homepage : : : : : : : : : :

.

. :

. . manufacturer ALBEMARLE CORPORATION Dr. M. Hardy 30.11.2004 451 Florida Street 70601 Baton Rouge, Louisiana United States 225-388-7616 225-388-7046
marcia-hardy@albemarle.com

11.01.2005

1.0.2 LOCATlON OF PRODUCTlON SITE, IMPORTER OR FORMULATOR

3.0.3 IDENTRY OF RECIPIENTS

1.0.4 DETAlLS ON CATEGORYPMPLATE

l-t.0 SUBSTANCE iDENTlFlCATlON

IUPAC Name Smiles Code Molecular formula Molecular weight Petrol class : : : : : 30.11.2004

1.7.1 GENERAL SUBSTANCE

Purity type : Substance type : Physical status : Purity : Colour : Odour : Attached document : HEXABROMOCYCLODODECANE clcc(Br)c(Br)ccc(Br)c(Br)ccc(Br)c2(Br) C12H18Br6 641.7

INFORMATlON

typical for marketed organic solid >= 96

- 99.9 % w/w white odorless substance

COMPOSITION OF THE COMMERCIAL HEXABROMOCYCLODODECANE (HBCD) PRODUCT
The commercial hexabromocyclododecane (HBCD) product contains three stereoisomers. The three isomers and gamma based on their order column. The predominant isomer stereoisomer. Based on analyses 2188 are typically referred to as alpha, beta of elution from a reverse phase HPLC

inthe commercial product is the gamma performed insupport of toxicology

## 1. General Information Id 25637-99-4 Date 31.03.2005

studies (described below), the commercial product consists of 80-85% gamma, 8-9% alpha, and 6% beta. The major impurity is tetrabromocyclododecene.
One manufacturer also produces a product of approximately 90% gamma HBCD content. However, the majority of the global market consists ofthe 80-85% gamma product.
The American Chemistry Council’s Brominated flame Retardant Industry Panel (BFRIP) sponsored a number of tests investigated the potential effects of HBCD. These tests were performed over the course of eight years. The test article used inthese tests was acomposite composed of equal parts ofthe members’ commercial HBCD product. BFRIP members are Albemarle Corporation, Dead Sea Bromine Group and Great Lakes Chemical Corporation. Each test article was characterized inaccordance with Good Laboratory Practices gLidelines. The composition of the test article used inthe various tests ranged from approximately 90 to 100% HBCD (Table 1). The impurities, when present, were those typically observed, e.g. tetrabromocyclododecene, isobutanol, and other unidentified compounds.
Tests performed from 2001-2003 typically utilized atest article of 99.9% purity (Table 1). The stereoisomer content of this test article was approximately 85% gamma, 9% alpha and 6% beta. Test article used in earlier years was comparable with a slightly lower gamma content (80% gamma, 8% alpha and 6% beta) (Table 1).

Table 1. COMPOSITION OF HBCD USED AS TEST ARTICLE IN BFRIP-SPONSORED STUDIES (all studies performed using a composite of equal parts ofAlbemarle Corporation, Dead Sea Bromine Group and Great Lakes Chemical Corporation commercial product) Study Year Mean Isomer Content (Area %) Isomer Sum (Purity) Alpha Beta Gamma
Validation of Water Solubility Analytical Method 1997 8.5 6.0 79.1 93.6 Water Solubility 1997 8.5 6.0 79.1 93.6 Log Kow 1997 8.5 6.0 79.1 93.6 Vapor Pressure 1997 8.5 6.0 79.1 93.6 Fish LC50 1997 8.5 6.0 79.1 93.6 Daphnia EC50 1997 8.5 6.0 79.1 93.6 Freshwater Algae EC50 1997 8.5 6.0 79.1 93.6 Daphnia Chronic 1998 8.5 6.0 79.1 93.6 Fish Early Life Stage 2001 9.4 6.3 84.3 100 Fish Bioconcentration+ 2000 6.4 4.5 79.1 90 Rat 28 Day 1996 8.5 6.0 79.1 93.6 Rat 90 Day 2001 8.9 6.6 84.5 99.99 Rat Developmental+ 3188

## 1. General Information Id 25637-99-4 Date 31.03.2005

1999 6.4 4.5 79.1 90 Guinea Pig Maximization 1997 8.5 6.0 79.1 93.6 Mouse LLNA* 2003 8.68 6.12 85.19 99.99 Chromosome Aberration 1996 8.5 6.0 79.1 93.6 In vivo Micronucleus 2000 8.9 6.6 84.5 99.99 Earthworm Survival & Reproduction* 2002 8.68 6.12 85.19 99.99 Terrestrial Plant: Seedling Emergence and Growth* 2002 8.68 6.12 85.19 99.99 Sediment Organism* 2003 8.68 6.12 85.19 99.99 Ready Biodegradability 1996 8.5 6.0 79.1 93.6 Sludge Respiration Inhibition* 2003 8.68 6.12 85.19 99.99 Soil Microcosm Biodegradation* 2003 8.68 6.12 85.19 99.99 Sediment Microcosm Biodegradation* 2003 8.68 6.12 85.19 99.99 Water Solubility of alpha, beta, gamma isomers 2004 7.67 5.15 83.04 95.86 Marine Algae, EC50 2004 7.67 5.15 83.04 95.86 *Results from reanalysis using stablilized THF; Initial analysis indicated 5.8% alpha isomer, 19.3% beta isomer, and 74.9% gamma isomer. +Impurities specified as TetraBCDodecene 0.7%, lsobutanol 0.1%; Other Unknowns 9.2%
Flag

. - Critical study for SIDS endpoint 11.01.2005

1.12 SPECTRA

1.2 SYNONYMS AND TRADENAMES

cyclododecane, hexabromo30.11.2004 HBCD
28.12.2004

1.3 IMPURrnES

Purity : typical for marketed substance CAS-No : EC-No : EINECS-Name : tetrabromocyclododecene Molecular formula : Value

. -

4188

## 1. General Information Id 25637-99-4 Date 31.03.2005

30.11.2004

1.4 ADDDIVES

1.5 TOTAL QUANTlTY

1.61 LADELUNG

1.6.2 CLASSIFICATION

1.6.3 PACKAGING

1.7 USE PATTERN

Type of use : industrial Category : Polymers industry
30.11.2004

1.7.1 DETAfLED USE PATTERN

Industry category : 11 Polymers industry Use category : 22 Flame-retardants and fire preventing agents Extra details on use category : Polymer processing No extra details necessary Emlssion scenario document : not available Product type/subgroup

. . Tonnage for Application . Year Fraction of tonnage for application Fraction of chemical in formulation Production

. . . . Formulation : : Processing : : Private use

. . Recovery

. . Attached document : HBCD is used solely as aflame retardant. Its primary application is in extruded (XPS) and expanded (EPS) polystyrene foam that is used as thermal insulation inthe building industry. HBCD is highly efficient inthis application so that very low levels are required to reach the desired flame retardancy. Typical HBCD levels in EPS are 0.67% and inXPS 2.5%. At present, HBCD is the only suitable flame retardant for these applications. Any other flame retardant would likely need higher load levels inthe polystyrene foam.
A secondary, though important, application of HBCD is as a flame retardant for upholstery textiles. In this application, HBCD is applied to the back of the upholstery fabric encapsulated ina polymer. Typical HBCD levels in the polymer backcoat are 6-l 5%. The potential exposure and hazard to consumers associated with this use were reviewed recently by the U.S. 5188

## 1. General Information Id 25637-99-4 Date 31.03.2005

National Research Council (D. Gardner and B. Walker, Chair, Toxicological Risks of Selected Flame Retardants, 2000, National Academy Press, Washington, D.C.; http:/www.nap.edu). The review found that direct exposure to the consumer was likely to be minimal, that the hazard index was less than 1for all exposure routes (e.g. not likely to pose a health hazard), and that no further research was needed for assessing consumer health risks from HBCD.
A very minor application for HBCD is invideo or audio equipment housings where V-2 levels of flame retardancy are acceptable. HBCD is not used to flame retard electronic housings (e.g. television sets) that must meet the higher V-O flame retardancy standard. 28.12.2004

1.7.2 METHODS OF MANUFACTURE

Origin of substance : Synthesis

. Type . 30.11.2004

1.8 REGULATORY MEASURES

1.8.1 OCCUPATtONAL EXPOSURE LIMIT VALUES

1.8.2 ACCEPTABLE RESIDUES LEVELS

1.8.3 WATER POLLUTlON

1.8.4 MAJOR ACCIDENT HAZARDS

1.8.5 AIR POLLUTION

1.8.5 LISTINGS E.G. CHEMICAL INVENTORIES



### Type

. - EINECS Additional information : TSCA; Japan
28.12.2004

1.9.1 DEGRADAT~ON!TRANSFOR~I~AT~ON PRODUCTS

1.9.2 COMPONENTS

6188

## 1. General Information Id 25637-99-4 Date 31.03.2005

1.10 SOURCE OF EXPOSURE

1.11 ADDITIONAL REMARKS

1.12 LAST LITERATURE SEARCH

Type of search

- External Chapters covered I Date of search : 15.12.2004
Attached document : A search for alldocuments containing the word “hexabromocyclododecane” was performed using PubMed. The search produced 20 hits. 15.12.2004

I.13 REVIEWS

7188

## 2. Physico-Chemical

2.1 MELTlNG POINT

Decomposition Sublimation Method Year GLP Test substance
Attached document

28.12.2004

2.2 BOILING POINT

2.3 DENSITY

2.3.1 GRANULOMEIRY

Type of distribution Precentile
Attached document

27.12.2004

2.4 VAPOUR PRESSURE

Value Decomposition Method Year GLP Test substance
Method Remark

Reliability Flag 28.12.2004 

## Data Id 25637-99-4 Date 31.03.2005

: yes, at ca. 269

- 279 “C : :

. . : no : as prescribed by 1.1

- 1.4

: The commercial HBCD has a melt range of 160-165 degrees Centigrade.
TGA analysis ofthe commercial HBCD product indicates 50% weight loss at 269 degrees Centigrade and 90% weight loss at 274 degrees Centigrade. (1)

: other

. .

: HBCD used in extruded or expanded polystyrene (predominant application) is sold either as granules (> 1 mm) or large particles (mean diameter 110 microns).
For use intextile applications (lesser application), HBCD is ground to afine particle size inorder to ease its dispersion incoatings. The average particle size for textile-grade-HBCD is 1.98 microns.

: = .0000627

- hPa at 21 “C : no : OECD Guide-line 104 “Vapour Pressure Curve” : 1997 : yes : as prescribed by 1.1

- 1.4

. - US EPA OPPTS 830.7950. Determined using the spinning rotor gauge. : Sponsor: American Chemistry Council (ACC) Brominated Flame Retardant Industry Panel (BFRIP) Sponsor: American Chemistry Council (ACC) Brominated Flame Retardant Industry Panel (BFRIP) : (1) valid without restriction : Risk Assessment, Critical study for SIDS endpoint (2)

8188

## 2. Physico-Chemical Data Id 25637-99-4 Date 31.03.2005

2.5 PARTlTlON COEFFICIENT

Partition coefficient : octanol-water Log pow : = 5.625

- at 25 “C pH value : - Method : other (measured) Year : 1997 GLP : yes Test substance : as prescribed by 1.1

- 1.4
Method : US EPA OPPTS 830.7560. Gnenerator column method. Remark : Sponsor: American Chemistry Council Brominated Flame Retardant Industry Panel (BFRIP). Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 30.11.2004

2.6.1 SOLUBlLtTY IN DIFFERENT MEDIA
Solubility in . Water Value . = 3.4

- other: ug/L at 25 “C pH value . - concentration : at “C Temperature effects

. . Examine different pol. : PfQ

. . at 25 “C Description . . Stable : 

## yes Deg. product

. . Method : GCD Guide-line 105 Year

. . 1997 GLP : Yes Test substance

. . as prescribed by 1.l

- 1.4
Remark : Sponsor: ACC Brominated Flame Retardant Industry Panel Attached document . HBCD’s water solubility using the generator column/column elution method (EPA 796.1860)lOECD 105) was determined to be 3.4 ug/L at 25 degrees C. The analytical method, HPLC using a UV dectector, was validated prior to use (Kendall and Nixon. Analytical method verification for the determination of hexabromocyclododecane (HBCD) inwell water. Final Report. Wildlife International LTD. Project Number: 439C107. Wildlife International LTD, Easton, MD). This method, the most sensitive available at the time of study performance, was capable of quantitating the gamma isomer, only. UV-active substances, whose peaks could not be resolved, interfered with quantifying the alpha and beta content. The solubility of HBCD was therefore based on that of the gamma isomer, which comprised -80% of the test article.
The results of this water solubility study were used to set dose levels for aquatic studies performed between 1997 and 2001, and sponsored by ACC Brominated Flame Retardant Industry Panel. Reliability (1) valid without restriction Flag Critical study for SIDS endpoint 16.12.2004 (3) Solubility in Water Value ca. 2.08

- 48.8 other: ug/L at 20 “C pH value - concentration at “C 9188

## 2. Physico-Chemical Data Id 25637-99-4 Date 31.03.2005

Temperature effects .

. Examine different pol. : PKa : at 25 “C Description .

. Stable : yes Deg. product : no Method : OECD Guide-line 105 Year : 2004 GLP : yes Test substance : as prescribed by 1.1

- 1.4
Remark : Sponsor: ACC Brominated Flame Retardant Industry Panel Attached document : HBCD: WATER SOLUBILITY OF THE THREE DIASTEROMERS
A study, which was performed according to current OECD/EPA guidelines and Good Laboratory Practices (GLPs), detem-rined HBCD’s water solubility to be 3.4 ug/L using a generator column with HPLC/UV as the analytical method (Stenzel and Markley, 1997). The test article was a composite of the 3 manufacturers’ commercial product and its composition was 79.1% gamma, 6.0% alpha and 8.5% beta. The best analytical method available atthat time was used; e.g. HPLC with a UV detector. That method was able to quantify the gamma isomer. UV-active substances, whose peaks could not be resolved, interfered with quantifying the alpha and beta content. Thus, the solubility of the commercial product was based on that of the gamma isomer. This was a reasonable approach, given that the technical product was largely composed (-80%) of the gamma isomer.
In late 2003, further studies were undertaken to determine HBCD’s potential effects on the marine algae, Skeletonema, inorder to clarify results reported inWalsh et al., 1987. Preliminary work for the 2003 algae study (Desjardins et al. 2003) investigated whether HBCD exposures in marine media would be best conducted utilizing the water accommodated fraction (WAF) or eluates produced from a generator column. The analytical method utilized was HPLCYMS, an evolution of the original method that allowed quantification of all3 isomers. The WAF preparations had serious problems with HBCD particulates that could not be resolved in a consistent manner by filtering or centrifugation. Thus, itwas concluded the WAF method was not appropriate. The HBCD eluate from the generator column solved the particulate problem, but also revealed that the water solubility of the alpha and beta isomers was significantly different than that of the gamma isomer. The water solubility of the gamma isomer inthe marine media was nearly equivalent to that determined in 1997 with reagent water. The solubility of the alpha and beta isomers was approximately 47 and 16 ug/L, respectively. A standard GLP solubility study (MacGregor and Nixon. 2004) using reagent water confirmed these results. Thus, under the conditions of the generator column, HBCD’s total water solubility (expressed as the sum of the three isomers) was 65.6 ug/L, and that of the gamma, alpha and beta isomers was 2.08, 48.8 and 14.7 uglL, respectively. Desjardins D, MacGregor J and Krueger H. 2003. Draft Report: Hexabromocyclododecane (HBCD): A 72-Hour Toxicity Test with the Marine Diatom (Skeletomema costatum). Wildlife International Project Number: 439A-125. Wildlife International, Ltd. Easton, MD. MacGregor J and Nixon W. 2004. Final Report. Determination of Hexabromocyclododecane (HBCD) Diastereomers using a Generator Column Method. Wildlife International Project Number: 439G138. Wildlife International, Ltd. Easton, MD.
10188

## 2. Physico-Chemical Data Id 25637-99-4 Date 31.03.2005

Stenzel J. and Markley B. 1997. Final Report. Hexabromocyclododecane (HBCD): Determination of the Water Solubility. Wildlife International Project Number: 439C-105. Wildlife International, Ltd. Easton, MD. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 20.12.2004 (4) Deg. product

. Method : other Year : 2000 GLP : no data Test substance : as prescribed by 1.1

- 1.4
Attached document : Atechnical data sheet from Albemarfe Corooration (accessed 12/2002) reports the following solubilites for HBCD: ’ Solubility (weight % at 25°C) Water..

.................. c 0.01 Acetone

.................. 6.6 Methanol

.................. 0.15 Toluene..

.................. 6.40 n-Pentane..

................ 0.01 lsopentane

................ 0.01 Cyclopentane

............... 0.05 Styrene..

.................. 

### a.00 Chlorobenzene..

............ 2.60 Methylene dibromide..

...... 3.60 Dimethyl formamide

....... 33.90 27.12.2004

2.6.2 SUl?FACE.TENSION

2.7 FLASH POINT

2.8 AUTO FLAMMABILITY

2.9 FLAMMABILITY

2.10 EXPLOSIVE PROPERTIES

2.11 OXIDIZING PROPERTlES

2.12 DlSSOClATlON CONSTANT

2.13 VISCOSITY

2.14 ADDlTlONAL REMARKS



### iii88

## 3. Environmental Fate and Pathways Id 25637-99-4

3.1.1 PHOTODEGRADAT’lON

3.1.2 STABlLlTY IN WATER

3.1.3 STABILITY IN SOIL


### Type : Radiolabel : Concentration : Soil temperature : Soil humidity : Soil classification

- Year I Content of clay. : Content of silt

. . Content of sand

. . Organic carbon

. . PH : Cation exch. capacity

. . Microbial biomass : Dissipation time DT50

. . DT90

. . Dissipation

. . Deg. product : Method : Year : GLP : Test substance : Attached document : Date 31.03.2005

other: laboratory, aerobic no .025 mg/kg 20 “C 20 other: % USDA 2003 =16- % =20- % =64- % =1.8- %

= 6.4

- = 19.2 meq/lOO g soil dry weight

= 274 other: ug/g dry weight soil

= 63

- day(s)

- = 75

- % after 119 day(s)
other: OECD 307 2003 yes as prescribed by 1.l

- 1.4 HBCD Transformation inAerobic and Anaerobic Soil Microcosms (Sponsor: ACC Brominated Flame Retardant Industry Panel and Dow Chemical Company)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. Its composition was 8.68% alpha isomer, 6.12% beta isomer and 85.19% gamma isomer. This study was performed according to Good Laboratory Practices and OECD 307.
The transformation of HBCD was determined inaerobic and anaerobic soils based on the Organization for Economic Co-Operation and Development (OECD) Test Guideline 307 “Aerobic and Anaerobic Transformation in Soil.” Soil microcosms were prepared by adding asandy loam surface soil to serum bottles sealed with Teflon coated septa. Aerobic microcosms were prepared by adjusting the soil moisture to 20% (by weight) and periodically exchanging the headspace of the microcosms with ambient air to replenish oxygen. The microcosms were pre-incubated at 20 f 1“C for 35 days. Anaerobic microcosms were prepared inan anaerobic atmosphere (70% N2, 28% C02, and 2% H2) by flooding the soil with water and pre-incubating the microcosms at 23 f 1“C for 43 days to allow low redox (e.g., methanogenic) conditions to develop. HBCD was then added to microcosms at a nominal concentration of 25 rig/g (soil dry weight), together with activated sludge (5 mg/g, dry weight basis) from a municipal wastewater treatment plant to simulate sludge land treatment applications. Biologically inhibited (abiotic) controls were prepared by 12188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

steam sterilization prior to the addition of HBCD. Microcosms were incubated inthe dark at 20 f 1“C for 119 days. The concentration of HBCD inthe microcosms was determined at selected time intervals utilizing high performance liquid chromatography-mass spectrometry (LCMS). Aerobic microcosms were analyzed on days 0, 1,7,21,48, 65, and 119, while anaerobic microcosms were analyzed on days 0, 1,7,21, 56, 91, and 119. HBCD concentrations decreased over time in both the aerobic and anaerobic soils. HBCD concentrations decreased 75% over 119 days inthe viable aerobic soil microcosms, compared to a 3% decrease inthe abiotic controls, indicating that biological processes were responsible for most of the losses observed. Under anaerobic conditions, HBCD concentrations decreased 92% over 21 days inthe viable microcosms compared to a less than 1% decrease inthe abiotic controls. Pseudo-first order kinetics rate constants for the biotransformation of HBCD were determined by subtracting the abiotic rate constant from the viable rate constant. Biotransformation half-lives for HBCD were determined to be 63 and 6.9 days inthe aerobic and anaerobic soils, respectively. Brominated degradation products were not detected inthe soil or inthe headspace of the microcosms.
The purpose of this study was to determine the environmental lifetime of HBCD under realistic environmental conditions. Laboratory microcosms were used to evaluate the transformation of HBCD inaerobic and anaerobic soils. Soil degradation processes are expected to play a major role in determining the environmental lifetime of HBCD since, upon release into the environment, the majority of HBCD is likely to partition into the soil or sediment compartments. In this study, HBCD loss was observed in both viable and abiotic soil microcosms although the rates were appreciably faster inthe viable soils. Biologically mediated transformation processes (i.e., biotransformation) accelerated the rate of loss of HBCD when compared to the biologically inhibited (i.e., heat-treated) soils. No brominated degradation products were observed in either system. Limited information is available for the reactions of HBCD inthe environment. However, the aerobic degradation and mineralization of a similar type of cyclic, aliphatic halogenated fire retardant, FR-651A (mixture of pentabromochlorocyclohexane, tetrabromo-dichlorocyclohexane, and tribromotrichlorocyclohexane) was observed. A soil half-life of -11 days based upon disappearance of 14 C-FR-651A from soil was reported. Complete degradation of 14C-FR-651 A was also observed with mineralization half-life on the order of 93 days.
An examination of the reactions of brominated aliphatic compounds that contain structural features similar to HBCD can also provide insight into the possible reaction pathways available for HBCD. Brominated aliphatic compounds are known to be susceptible to both hydrolytic and nuclelophilic attack. The reactivity of halogenated aliphatic compounds increases inthe order of F< Cl < Br. For example, the hydrolysis half-lives at pH 7and 25 “C for methyl fluoride, methyl chloride, and methyl bromide are 1.l x 104, 340, and 20 days, respectively. At environmental pH’.s, neutral and base catalyzed hydrolysis reactions are most important, and depend on the degree and patterns of halogen substitution. Sulfur based nucleophiles can react rapidly with halogenated aliphatic compounds, thereby reducing their lifetimes inthe environment. The halflife of 1-bromohexane inwater at 25 “C was reduced from 20 days to approximately one day in5 mM HS -or 0.07 mM polysulfide (Sx 2- ). Similarly, the half-life of 1,2-dibromoethane in water at 25 “C was reduced from 1,000 days to 4 days inthe presence of 5 mM HS-. Sulfide and polysulfides would be expected to be present insoils 13188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

at low redox potentials. Thus, reaction of HBCD with these sulfur-based nucleophiles may partly explain the loss of HBCD inthe soil studies.
Since HBCD contains three pairs of vicinal bromine atoms, the transformation of simple aliphatic compounds containing vicinal bromine atoms may provide insight into possible reaction pathways for HBCD. For example, 1,Bdibromoethane reacts with nucleophiles via both substitution and elimination reactions. Reaction with HS- via an “SN2” substitution reaction results inthe formation of HS-CH2-CH2-SH, while an elimination reaction under alkaline conditions results inthe formation of H2C=CHBr. A combination of elimination and substitution reactions can result inthe formation of a mixture of HO-H2-CH2-OH and H2C=CHBr. Similar mechanisms may be responsible for the loss of HBCD observed inthis study. Halogenated aliphatic compounds are susceptible to reductive dehalogenation reactions in natural reducing environments. The rates of reactions vary depending on the strength and electron affinity of the carbon-halogen bond and the stability of the carbon-radical species resulting from the initial electron transfer that occurs. Vicinal dehalogenation reactions are particularly important, and are dependent on the halogen atom. Half-lives for vicinal dehalqenation of 1,2-diiodoethane, 1,2-dibromoethane, and 1,2-dichloroethane are 0.6, 55, and 1.8 x 104 hours, respectively. The rapid disappearance of HBCD inthe anaerobic soil microcosms may be partly explained by reductive dehalogenation reactions.
Davis J, Gonsior S and Marty G. 2003. Evaluation Of Aerobic And Anaerobic Transformation Of Hexabromocyclododecane In Soil. Study Number 021082. Environmental Chemistry Research Laboratory. Toxicology & Environmental Research and Consulting. The Dow Chemical Company. Midland, Ml. Reliability (1) valid without restriction Flag Critical study for SIDS endpoint 12.01.2005 (5) Type : other: laboratory, anaerobic Radiolabel : no Concentration

. . .025 mg/kg Soil temperature

. . 20 “C Soil humidity : 20 other: % Soil classification : USDA Year

. . 2003 Content of clay : =16- % Content of silt : =20- % Content of sand : =64- % Organic carbon . = 1.8- % PH . = 6.4

- Cation exch. capacity

. . = 19.2 meq/lOO gsoil dry weight Microbial biomass : = 274 other: ug/g dry weight soil Dissipation time DT50 = 6.9

- day(s) DT90 Dissipation = 92

- % after 21 day(s) Deg. product Method other: OECD 307 Year 2003 GLP yes Test substance as prescribed by 1.1

- 1.4 Attached document Transformation inAerobic and Anaerobic Soil Microcosms (BFRIP and Dow Chemical Company) 14188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. Its composition was 8.68% alpha isomer, 6.12% beta isomer, and 85.19% gamma isomer. This study was performed according to Good Laboratory Practices and OECD 307.
The transformation of HBCD was determined inaerobic and anaerobic soils based on the Organization for Economic Co-Operation and Development (OECD) Test Guideline 307 “Aerobic and Anaerobic Transformation in Soil.” Soil microcosms were prepared by adding asandy loam surface soil to serum bottles sealed with Teflon C coated septa. Aerobic microcosms were prepared by adjusting the soil moisture to 20% (by weight) and periodically exchanging the headspace of the microcosms with ambient air to replenish oxygen. The microcosms were pm-incubated at 20 f 1“C for 35 days. Anaerobic microcosms were prepared inan anaerobic atmosphere (70% N2,28% C02, and 2% H2) by flooding the soil with water and pre-incubating the microcosms at 23 f 1“C for 43 days to allow low redox (e.g., methanogenic) conditions to develop. HBCD was then added to microcosms ata nominal concentration of25 rig/g (soil dry weight), together with activated sludge (5 mg/g, dry weight basis) from a municipal wastewater treatment plant to simulate sludge land treatment applications. Biologically inhibited (abiotic) controls were prepared by steam sterilization prior to the addition of HBCD. Microcosms were incubated inthe dark at 20 f 1“C for 119 days. The concentration of HBCD inthe microcosms was determined at selected time intervals utilizing high performance liquid chromatography-mass spectrometry (LCMS). Aerobic microcosms were analyzed on days 0, 1,7,21,48, 65, and 119, while anaerobic microcosms were analyzed on days 0, 1,7,21,56, 91, and 119. HBCD concentrations decreased over time in both the aerobic and anaerobic soils. HBCD concentrations decreased 75% over 119 days inthe viable aerobic soil microcosms, compared to a3% decrease inthe abiotic controls, indicating that biological processes were responsible for most of the losses observed. Under anaerobic conditions, HBCD concentrations decreased 92% over 21 days inthe viable microcosms compared to a less than 1% decrease inthe abiotic controls. Pseudo-first order kinetics rate constants for the biotransformation of HBCD were determined by subtracting the abiotic rate constant from the viable rate constant. Biotransformation half-lives for HBCD were determined to be 63 and 6.9 days inthe aerobic and anaerobic soils, respectively. Brominated degradation products were not detected inthe soil or inthe headspace of the microcosms.
The purpose of this study was to determine the environmental lifetime of HBCD under realistic environmental conditions. Laboratory microcosms were used to evaluate the transformation of HBCD inaerobic and anaerobic soils. Soil degradation processes are expected to play a major role indetermining the environmental lifetime of HBCD since, upon release into the environment, the majority of HBCD is likely to partition into the soil or sediment compartments. In this study, HBCD loss was observed in both viable and abiotic soil microcosms although the rates were appreciably faster inthe viable soils. Biologically mediated transformation processes (i.e., bfotransformation) accelerated the rate of loss of HBCD when compared to the biologically inhibited (i.e., heat-treated) soils. No brominated degradation products were observed in either system. Limited information is available for the reactions of HBCD inthe environment. However, the aerobic degradation and mineralization of a similar type of cyclic, aliphatic halogenated fire retardant, FR-651 A (mixture of pentabromochlorocyclohexane, tetrabromo-dichlorocyclohexane, and 15188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

tribromotrichlorocyclohexane) wasobserved.A soilhalf-life of -11 days basedupondisappearanceof 14C-FR-651A fromsoilwas reported. Completedegradation of 14C-FR-651Awas alsoobservedwith mineralization half-life onthe orderof93 days.
An examination ofthe reactionsof brominatedaliphatic compoundsthat contain structural featuressimilarto HBCDcan alsoprovide insightinto the possiblereaction pathways availablefor HBCD. Brominatedaliphatic compoundsare knownto besusceptibleto both hydrolytic and nuclelophilic attack. The reactivity of halogenatedaliphatic compoundsincreasesinthe orderof F cCl c Br. Forexample,the hydrolysis half-lives at pH 7and25 “Cfor methylfluoride, methylchloride, and methyl bromideare 1.l x 104, 340, and20 days, respectively. At environmental pH’s,neutral andbase catalyzed hydrolysis reactionsare mostimportant, anddependonthe degree andpatterns of halogensubstitution. Sulfur basednucleophilescan react rapidlywith halogenatedaliphatic compounds,thereby reducingtheir lifetimesinthe environment. The halflifeof 1 -bromohexane inwaterat25 “Cwas reducedfrom20daysto approximately one day in5 mM HS- or0.07 mM polysulfide (Sx 2- ). Similarly,the half-life of 1,Pdibromoethane in waterat25 “C wasreducedfrom 1,000daysto4 days inthe presenceof5 mM HS-. Sulfide and polysulfideswould beexpected to bepresentinsoils at low redox potentials.Thus, reactionof HBCDwiththesesulfur-based nucleophilesmaypartly explainthe lossof HBCD inthe soilstudies. Since HBCD containsthree pairsofvicinal bromineatoms,the transformation ofsimplealiphaticcompoundscontainingvicinal bromine atomsmay provide insightinto possiblereaction pathwaysfor HBCD. For example, 1,Bdibromoethane reactswith nucleophilesvia both substitution andeliminationreactions. Reactionwith HS-via an ‘SN2” substitution reaction resultsintheformation of HS-CHBCHFSH, whileanelimination reaction underalkalineconditions resultsintheformation of H2C=CHBr.A combinationofeliminationandsubstitution reactionscan resultinthe formation ofa mixture of HO-H2-CH2-OH and H2C=CHBr. Similar mechanismsmaybe responsibleforthe lossof HBCD observed inthis study. Halogenatedaliphaticcompoundsaresusceptibleto reductive dehalogenationreactions innatural reducingenvironments.The ratesof reactionsvary dependingonthestrength and electron affinity ofthe carbon-halogen bondandthestabilityofthe carbon-radicalspecies resultingfromthe initialelectrontransferthat occurs.Vicinal dehalogenation reactionsare particularly important, andare dependenton the halogenatom. Half-livesforvicinal dehalogenationof 1,2-diiodoethane, 1,2-dibromoethane, and 1,2-dichloroethaneare 0.6,55, and 1.8x 104 hours, respectively. The rapiddisappearanceof HBCDinthe anaerobicsoil microcosmsmaybe partly explainedby reductive dehalogenation reactions.
DavisJ, Gonsior SandMarty G. 2003. Evaluation Of Aerobic And AnaerobicTransformation of Hexabromocyclododecane In Soil. Study Number021082. Environmental Chemistry ResearchLaboratory. Toxicology & Environmental Researchand Consulting.The DowChemical Company. Midland, Ml. Reliability : (1)valid without restriction Flag : Criticalstudy for SIDS endpoint 12.01.2005 (5)

06.12.2004

1618806.12.2004 

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

3.2.1 MONITORING DATA

Type of measurement : background concentration Media : other: biota, sewage treatment plant, sediment, landfill leachate Concentration -
. - Method .

. Attached document : HEXABROMOCYCLODODECANE ENVIRONMENTAL MONITORING: RESULTS OF ISOMER-SPECIFIC ANALYSIS AVAILABLE AS OF MAY 2004 Hexabromocyclododecane (HBCD) is used to flame retard extruded and expanded polystyrene that is then used as thermal insulation in buildings. A minor use is in upholstery textiles where it is applied as a backcoat on the fabric. The commercial HBCD product is composed ofthree stereoisomers that are designated alpha, beta and gamma indeference to their respective elution from a reverse phase column, The isomer content of the commercial product is typically 80-85% gamma, 8-9% alpha and 6% beta. The major impurity is tetrabromocyclododecene.
A laboratory fish bioconcentration study showed that the three stereoisomers were present in rainbow trout in rough approximation to that ofthe commercial product used as test article. A rat 90-day subchronic study on the commercial product showed that the alpha isomer predominated inadipose tissue with lesser quantities of the gamma and beta isomers. Given these apparent differences between species and isomers, the literature and unpublished data were reviewed for environmental monitoring studies that included HBCD (see Table below). Only those studies that utilized analytical methods allowing detection of the three HBCD isomers were included. Studies reporting only ‘total HBCD’ content were discarded. Results were separated by matrix: sewage treatment plants, sediment, landfill leachate, whole organisms, and specific tissues. The resulting dataset both as awhole and for any specific matrix/organism is small. Thus, caution should be used to guard against over interpretation of the results. Sewage Treatment Plants (STP) STPs appear to do a good job removing HBCD from the influent. Effluent from plants with detectable amounts of HBCD inthe influent typically had no detectable HBCD. HBCD appears to settle insludge; the gamma isomer typically represented some 50% ofthe total. Sludge from the UK and Ireland typically contained allthree isomers, but only one STP inthe Netherlands had detectable levels ofthe beta isomer insludge. For the 5 UK plants with measured influent, 1 had no detectable HBCD inthe influent, 3 had gamma but no alpha or beta inthe influent, and 1 had all3 isomers with beta predominating. Inthe Netherlands, for the 5 plants measured, 3 had alpha inthe influent but not beta or gamma, one had predominantly gamma with lesser alpha, and one had no detectable HBCD. Conclusion: STPS perform their function in removing HBCD from the influent. Effluents from plants receiving HBCD typically contained no detectable HBCD. Gamma typically made up approximately 50% ofthe total insludge; e.g. the gamma predominated. The predominant isomer in the influent appears to vary with country (gamma in UK, alpha inthe Netherlands)/ however, the dataset is extremely small and variable so this should be viewed with caution.

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## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

Sediment In those sediments with detectable HBCD, the gamma isomer was detected with greatest frequency (9 out of 27 sediment samples from the Scheldt and Dutch Rivers had gamma as the predominant isomer and many of these had detectable levels of gamma only). Two sediments out of 27 had roughly equal levels of gamma and beta, one had roughly equal levels of alpha and gamma, and one had alpha only. Conclusion

- the HBCD isomer most often detected insediment was gamma. Landfill Leachate Samples from the UK and Ireland had no detectable HBCD. Eleven samples of landfill leachate originating inthe Netherlands were analyzed. Four contained no detectable HBCD. In the seven where HBCD was detected, the gamma isomer was the predominant and most frequently detected isomer. The beta isomer was detected inonly one of the samples from the Netherlands.
Whole Organisms Both alpha and gamma were detected inall Lake Ontario trout analyzed (5/5). Beta was not detected inany. Composites of 3different feeder fish from Lake Ontario also had detectable levels of both alpha and gamma, but no beta. Composites of 3 invertebrates, analyzed separately by species, from Lake Ontario also had detectable levels of both alpha and gamma, but no beta. Alpha was present inthe highest quantities inall Lake Ontario organisms. North Sea whelk, sea star and hermit crabs had no detectable levels of HBCD. Conclusion -The alpha and gamma isomers were detected with the same frequency infreshwater species of Lake Ontario. The alpha isomer typically represented approximately 80% of the total. Analysis of whole organism, as opposed to individual tissues (see below), appears to generate the most consistent data. Specific Tissues Liver, bird. The alpha isomer was most frequently detected (and the predominant isomer) in cormorant liver (n=5). Liver, marine mammal. One marine mammal liver contained alpha but not beta or gamma, and one contained no detectable HBCD (n=2). Thus, no conclusion can be suggested. Muscle, fish. The dataset for Whiting is too small and variable to draw any conclusion. Whitefish (Switzertand) contained alpha, but no beta or gamma. Muscle, bird. The alpha, beta and gamma isomers were detected with roughly equal frequency. Egg, bird. The alpha isomer was most frequently detected. The numbers of eggs with detectable quantities of beta and gamma were roughly equal. Blubber. The alpha isomer was the most frequently detected and typically inthe highest quantity. Only one sample out of 11 had detectable amounts of all3 isomers. Eel. The alpha isomer was the most frequently detected and typically made up the largest proportion of the total. However, a reasonable number of samples also contained gamma, and infew cases, the alpha and gamma levels were approximately equal. Conclusion. This dataset is extremely small and highly variable. Continued analysis of liver, muscle and eel does not appear justified. Egg and blubber may be good indicators for birds and marine mammals, and thus continued analysis of these tissues may prove useful.
Textile Plants Located in Europe The gamma isomer was the predominant HBCD isomer detected inwaste water from 4 of 5textile plants. In 1ofthese 5 plants, alpha and gamma were present inapproximately equal quantities. The gamma isomer 18188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

predominatedinsoilsurroundingthe 5 plants.
SUMMARY The publishedliterature and unpublisheddatawere reviewedfor environmental monitoringresultspertainingto HBCD. Onlythosestudies that includedspecificanalysisfor the alpha, beta andgammaHBCD stereoisomerswere included. Environmental matrixes reporting resultsfor allthree stereoisomersincludedsewagetreatment plants,sediment,whole organisms,specifictissues,andtextile plantwastewater, andtextile plant soil. The gammaisomerpredominatedinsewagetreatment plant sludge, sediment, landfillleachate,textile plantwastewater andtextile plantsoil. The alphaisomerwasthe predominant isomerdetected onthe analysisof whole organisms(laketrout, feederfish, 3freshwater invertebrates). Analysis of individualtissuesproducedvarying results. Ofthe individual tissuesanalyzed, avian egg andmarinemammalblubberappearedto producethe mostconsistent results. The alphaisomerwasthe most frequently detected isomerinboth. Continued monitoringofeggand blubbermaybeofvalue, but othertissuetypes are not recommended. The mostconsistentdata isapparently generated onthe basisofwhole organismanalysis. Sewagetreatment plantsareeffective inremoving HBCDfromthe influent- effluentsfrom plantsreceiving HBCDtypically contained nodetectable HBCD.

TABLE: HBCD ISOMER SPECIFIC ANALYSIS
A. InVariousTissues(Datafrom draft EURA 2003, Gerecke2003 andde Boer’sbirdstudy2004)
Liver (All ppbww) alpha beta gamma Cormarant (@kg ww) 21 3 2 % Lipidnotstated 6 1 2 33 1 1 7 0.3 0 2 0 0 H. Porpoise(rig/gww) 2400 0 4 36%lipid,n=l H. Seal0 b”“‘” 0 2% lipid, n=2or3
Muscle OR Fillet (All ppbww) Fish Whiting, UK(@kg ww) 506 247 283 % Lipidnotstaed 81 61 149 0 0 0 Whiting, Netherlands(rig/gww) 0 0 0 0.6% Lipid, n=3 Whitefish, Switzerland, (rig/g Iw) 210 0 0 2.6%., poolof 10 100 0 0 3.8%, poolof10 66 0 0 1.5%, poolof 10 54 0 0 7.2%, poolof 10 Avian Sparrow Hawk (ug/kgww) 61 0 0 2.6% Lipid 0 8.1 19 1.3 0 9.4 11 2.2 2.6 0 0 3.1 13 26 150 1

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## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

0 7.1 0 0.8 4.8 0 0 1 2.6 0 0 1.1 0 9.2 0 1 0 0 0 0 - 8.5% LIPID, n=60 Egg, Avian (@kg ww) Falcon 38 0 0 7 72 0 0 11.1 22 0 0 7.7 0 9.2 0 0.8 13 0 0 7.3 5.1 0 0 7.2 0 3.4 4.6 6.4 28 0 0 5.5 30 13 12 7.1 0 8.9 0 6.3 15 0 5.5 6.3 27 0 0 4.2 20 0 0 5.0 0 0 0 0.8-l 7.1; n=40
Blubber (All ppb ww) Porpoise (@kg ww) 298 302 317 % Lipid not stated, n=5 f 315 2 5 53 1 0 89 0.4 0 0 0 0 H. Porpoise (rig/g ww) 3500 0 0 90% Lipid, n=4 410 0 0 6400 0 0 800 0 0 H. Seal Ws ww) 2000 0 0 70% Lipid, n=2 0 0 0
Eel (whole body?) (ug/kg ww) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 25 5 11 25 0 8 5 0 0 0 0 0 22 21 1 2 0 0 5 0 0 3 0 0 0 0 0 0 0 0 52 0 12 36 0 8 28 0 7

20188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

a 0 0 14 0 0.4 17 0 15 2 0 0 4 0 0 a 0 IO

B. WHOLE ORGANISMS (Data from Tomy et al2004 and de Boer re North Sea) Alpha Beta Gamma %Lipid, n Lake Trout, Lake Ontario, whole fish (nglg ww), n=5 1 0 0.3 11 % lipid 0.4 0 0.07 14% lipid 1.2 0 0.26 9 % lipid 0.6 0 0.1 15 % lipid 3.8 0 0.7 14% lipid Rainbow Smelt, Lake Ontario, composites of whole fish (rig/g ww) 0.3 0 0.04 1.4% lipid, n=5 fish 0.2 0 0.04 1.6% lipid, n=5 fish 0.2 0 0.03 1% lipid, n=20 fish Slimy sculpin, Lake Ontario, composites of whole fish (rig/g ww) 0.1 0 0.02 0.9, r-r=15 0.4 0 0.1 3%, n=lO 0.4 0 0.1 2%, n=lO Alewife, Lake Ontario, composites of whole fish (rig/g ww) 0.15 0 0.02 3.5% lipid, n=5 0.12 0 0.01 5.5% lipid, n=5 0.08 0 0.01 1.5% lipid, n=5 Mysis, Lake Ontario, composite of whole organisms (rig/g ww) 0.07 0 0.02 3.1 % lipid, n>lOO 0.04 0 0.01 3.8% lipid, n>lOO Diporeia, Lake Ontario, composite ofwhole organisms (q/g ww) 0.06 0 0.03 1.7%lipid, n>lOO 0.05 0 0.02 l%lipid, n>lOO Plankton, Lake Ontario, composite of whole organisms (rig/g ww)
0.04 0 0.03 0.6% lipid 0.02 0 0 0.2% lipid Whelk, North Sea (rig/g ww) 0 0 0 2.4% 0 0 0 1.8 0 0 0 1.5 Sea Star, North Sea (q/g ww) 0 0 0 3.5% 0 0 0 7.5 0 0 0 7.6 Hermit Crab, North Sea (nglg ww) 0 0 0 7.4% 0 0 0 6.7 0 0 0 a.5 0 0 0 10.6 0 0 0 9 0 0 0 7.3 0 0 0 9.5 0 0 0 14.1 0 0 0 9.8 C. SEWAGE TREATMENT PLANTS (Data from de Boer et al., 2002; as reported indraft EU Risk Assessment 2003) Alpha Beta Gamma Burnham, UK lnfluent dissolved phase 21 t 88

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

7.9 12.5 3.2 lnfluent particulate phase 0 29.4 Sludge 132 Effluent dissolvedphase 0 0 Effluent particulate phase 0 0 Latchingdon, UK lnfluent dissolvedphase 0 0 9.1 lnfluent particulate phase 0 0 Sludge 205 Effluent dissolvedphase 0 0 Effluent particulate phase 0 0 Wickford, UK

lnfluent dissolvedphase 0 0 lnfluent particulate phase 0 0 Sludge 89.6 Effluentdissolved phase 0 0 Effluent particulate phase 0 0 S.Woodham Ferrers, UK lnfluent dissolvedphase 0 0 0 lnfluent particulate phase 0 0 Sludge 233 Effluentdissolved phase 0 0 Effluent particulate phase 0 0 Chelmsford,UK lnfluent dissolvedphase 0 0 lnfluent particulate phase 0 0 Sludge 541 Effluent dissolvedphase 0 0 Effluent particulate phase 0 0 STPl , Netherlands lnfluent 670 0 Sludge 0 Effluent 0 0 STP2, Netherlands lnfluent 3800 0 Sludge 0 Effluent 9 0 STPS, Netherlands lnfluent 40 0 Sludge 15 Effluent 0 0 5.1 STP4, Netherlands lnfluent 75 0 22/ 88 

## rig/L

0 458
0
0

2.3 321
0
0

4.6
0 112
0
0

0 547
0
0

4.3
0 897
0
0
0 0 0 0 0 8.7
0 0

180 @kg dw 666 uglkg dw
W- @kg dw

432

1245



## rig/L 0 @kg dw WL

48

5.4

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

Sludge 440 120 760 Effluent 0 0 0 STP5, Netherlands lnfluent 0 0 0 Sludge Effluent 1.4 0 0 STPG, Netherlands Sludge 7.6 0 20 STP7, Netherlands Sludge 3.5 0 3.6 STP8, Netherlands Sludge 7.4 0 13 STPS, Netherlands Sludge 0 0 0 STP1 0, Netherlands Sludge 0 0 0 Portlaoise, Ireland Sludge 280 349 604 Sludge 372 523 750 Clonmel, Ireland Sludge 3.9 0 149 Sludge 10.1 0.13 258 Cork, Ireland Sludge 2300 1800 3410 Population equivalents:

STP l-4,7 and 10 high treatment capacity, 200 000

- 750 000 STP5,6and8100000-150000 UK varying from 4 750, Latchingdon, to 143 000, Chelmsford.
D. SEDIMENT (uglkg ww) Scheldt Basin (Data from (de Boer et al., 2002) as reported indraft EU W AB G Warmebeek Achel-kluis 0 0 0 Moervaart Daknam 0 0 0 Benede Nete Duffel 0 0 0.5 Grote Beverdijk Lo-R. 0 0 0 ljzer Nieuwpoort 0 0 0.9 Durne Lokeren 00 0 Leie Wervik 00 0 Leie Wevelgem 00 0 Leie Oselgem 0 0 0 Leie St Martens 7 0 31 Scheldt Doel 00 0 Scheldt Grens 0 0 82 Scheldt Oudenaarde 180 60 710 Antwerp Kruisschansbr. 0 0 0 Scheldt Kastelt 0 0 5 Scheldt Kennedyt. 0.3 0 0 Dender Appels 0.3 0 0 Dender Ninovet 0 0 0.2 Dutch Rivers (Data from (de Boer et al., 2002) as reported indraft EU RA)
Waal Tiel 00 0 Rhine Lobith 04 4 Hollans Diep 00 10 Haringvliet West 0 0 6 Haringvliet East 0 0 0 Nieuwe Mervede 20 1 Meuse Eijsden 00 0 23188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

Meuse Keizersveer 3 0 0 Roer Vlodrop 0 0 0

UK 42 16-980 12-550 ppb dw Ireland 0.30 O-30 Dublin Bay, Ire 0 o-1 o-11 0 0 0 Norway o-9o-4 O-79 nglkg ww

E. Landfill LEACHATE (Data from draft EU Risk Assessment) UK 0 0 0 rig/L (dissolved) UK 0 0 0 rig/L (particulate) Netherlands O-7000 o-13 o-36000 ug/kg dw Ireland 0 0 0 rig/L (dissolved) Ireland 0 0 0 rig/L (particulate)

28.12.2004

28.12.2004

3.2.2 flELD STUDIES

3.3.1 TRANSPORT BETWEEN ENVIRONMENTAL COMPARTMENTS 

### Type : fugacity model level Ill Media : other: 1000 kg/hr to air, water, and soil Air

. . % (Fugacity Model Level I) : % (Fugacity Model Level I) Soil : % (Fugacity Model Level I) Biota

. . % (Fugacity Model Level ll/lll) Soil

. . % (Fugacity Model Level ll/lll) Method : other: EPlwin, Level Ill Fugacity Model Year : 2004
Attached document : HBCD’s partitioning inthe environment was estimated using the Level Ill Fugacity Model of EPlwin, V3.04. At default emissions of 1000 kg/hr to each of air, water and soil, HBCD was predicted to dsitribute primarily to soil (40.1%) and sediment (57.9%). Only 2.06% was predicated to partition to water with neglibilbe levels to air (0.000685%). The soil Koc was calculated by the model to be 2.25 x 10+7. The percent reacted inair, water, soil and sediment was estimated as 0.304, 3.26, 63.3 and 22.9%, respectively. The percent advected inair, water, soil and sediment was estimated as 0.0225, 6.77, 0, 3.8%, repectively. The overall half-lives (hr) estimated (based on Biowin (Ultimate) and Aopwin) inair, water, soil and sediment were 51.19, 1440, 1440 and 5760, respectively.
The model was also run with emissions solely to either air, water or soil. If released solely to air, HBCD was predicted to partition to soil (77.7%) and sediment (21.S%). Itwas expected to be 97.1% reacted insoil and sediment with only 2.91% undergoing advection. If released soley to
24188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

water, HBCD was predicted to partition to sediment (96.6% with 3.44% to water. The percent reacted would be 71.2% adn 26.8% advected. If released solely to soil, HBCD was predicted to partition 99.9% to soil with 100% reaction. 27.12.2004

3.3.2 DlSTRlBUTlON

27.12.2004

3.4 MODE OF DEGRADATlON IN ACTUAL USE

3.5 BlODEGRADATlON


### Type : aerobic lnoculum : activated sludge Concentration : 15 mg/l related to Test substance related to Contact time : 3 hour(s) Degradation

. . - (*) % after Result . Deg. product

. . Method : other: OECD 209 Year : 2003 GLP : yes Test substance : as prescribed by 1.1

- 1.4
Attached document : Activated Sludge Respiration Inhibition Test (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD (209) guidelines and Good Laboratory Practices.
HBCD’s effect on the respiration of activated sludge microorganisms was assessed using control, reference and treatment groups. The control group was used to determine the background respiration rate ofthe sludge and was not dosed with the test or reference substance. The reference group was dosed with 3,5-dichlorophenol, a known inhibitor of respiration, at nominal concentrations of 3, 15 and 50 mg/L. The test substance was dosed at a limit concentration of 15 mg/L. After an exposure period of

- three hours, the respiration rates of the test solutions were measured using a dissolved oxygen meter. The individual respiration rates ofthe two controls were 60.5 and 55.5 mg 02/Uhr. The difference between the two control respiration rates was 9.0% and was within the 15% difference limit established for the test. The validity of the test was further supported by the results from the 3,5-dichlorophenol reference group, which resulted inan EC50 of 5.2 mg/L and was within the 5 to 30 mg/L range considered acceptable for the test. An average of29.1 percent inhibition was observed inthe treatment group. Schaefer Eand Siddiqui A. 2003. Hexabromocylcododecane (HBCD): An Activated Sludge, Respiration Inhibition Test. Project Number: 439E-108A. Wildlife International, Ltd. Easton, MD. Reliability : (1) valid without restriction 25188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

Flag 29.12.2004 Type lnoculum Concentration
Contact time Degradation Result Deg. product Method Year GLP Test substance
Attached document

Reliability Flag 29.12.2004 Type lnoculum Contact time Degradation Result Deg. product Method Year GLP Test substance
Attached document : Critical study for SIDS endpoint

: aerobic : activated sludge, domestic : 7.7 mg/l related to Test substance related to

. 28 day(s) : = 0 - (k) % after 28 day(s) : under test conditions no biodegradation observed

. . no

.

. EPA OTS 796.3200

.

. 1996 : yes : as prescribed by 1.l

- 1.4

. . Closed Bottle Test For Biodegradability (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to OECD 301D, OPPTS 3200, and Good Laboratory Practices.
HBCD was tested for ready biodegradation ina28 day closed bottle test at a concentration of 7.7 mg/L by measuring dissolved oxygen uptake and expressing itas a percentage of the theoretical oxygen demand or chemical oxygen demand. No biodegradation was observed; the percent biodegradation was 0. Schaefer. Eand Haberlein. D.. 1996. Hexabromocyclododecane (HBCD): Closed Bottle Test. Project No.: 439E-102. Wildlife International‘Ltd. Easton, MD. : (1) valid without restriction

.

. Critical study for SIDS endpoint (7)

: aerobic : other: aerobic water/sediment microcosms : 119 day(s) : >= 90

- (k) % after 21 day(s)

.

. other: half-lives in2 river sediments = 11 and 32 days inaerobic microcosms. : not measured

.

. other: OECD 308

. 2003

.

.

. yes

.

. other TS

: Transformation inAerobic and Anaerobic Water/Sediment Microcosms (Sponsor: ACC Brominated Flame Retardant Industry Panel and Dow Chemical Company) The test article was a composite of 3 lots of commercial HBCD produced by Albemarfe Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. Its composition was 8.68% alpha, 6.12% beta, and 85.19% gamma. This study was performed according to Good Laboratory Practices and OECD Guideline 308.
The transformation of hexabromocyclododecane (HBCD) was determined inaerobic and anaerobic water/sediment microcosms based on the Organization for Economic Co-Operation and Development (OECD) Test 26 I 88

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

Guideline 308 “Aerobic and Anaerobic Transformation inAquatic Sediment Systems.” Laboratory batch microcosms were prepared with authentic water and sediment collected from two rivers inthe eastern United States. Aerobic microcosms were pre-incubated at20 f 1“C for 49 days and maintained by periodically exchanging the headspace ofthe microcosms with ambient air to replenish oxygen. Anaerobic microcosms were prepared in an anoxic atmosphere (70% N2,28% C02, and 2% H2). The microcosms were pre-incubated at 23 f 1“C for 43 to 44 days to allow the microcosms to stabilize. HBCD was then added to the microcosms at nominal concentrations ranging from 34 to 89 rig/g (sediment dry weight). Biologically inhibited (i.e., abiotic) controls were prepared by steam sterilization of the sediment/water mixture prior to the addition of HBCD. Microcosms were incubated inthe dark at 20 f 1“C for 119 days. The concentration of HBCD inthe microcosms was determined at selected time intervals inthe water and sediment phases utilizing high performance liquid chromatography-mass spectrometty (LC-MS). Aerobic microcosms were analyzed on days 0, 1,7,21,64,91, and 119, while anaerobic microcosms were analyzed on days 0, 1,7, 14,61 (or 62), 91, and 119. HBCD concentrations decreased over time in both the aerobic and anaerobic microcosms. HBCD concentrations inthe viable aerobic microcosms from both river systems decreased at least 90% within 21 days, while the corresponding decreases inthe abiotic controls ranged from 7to 62%. Disappearance of HBCD was observed in both the viable and abiotic anaerobic microcosms with the rate of loss more rapid inthe viable microcosms, with HBCD reaching non-detected levels within 7 days. In contrast, HBCD concentrations inthe abiotic controls decreased from 48 to 62% after 14 days. Pseudo-first order kinetics rate constants for the biotransformation of HBCD were determined by subtracting the abiotic rate constant from the viable rate constant. Biotransformation half-lives for HBCD inthe two river systems were determined to be 11and 32 days inthe aerobic microcosms and 1.l and 1.5 days inthe anaerobic microcosms. Brominated degradation products were not detected inthe sediment and water layers or inthe headspace of the microcosms.
The purpose ofthis study was to determine the environmental lifetime of HBCD under realistic environmental conditions. Laboratory microcosms were used to evaluate the transformation of HBCD inaerobic and anaerobic water/sediment microcosms. Sediment degradation processes are expected to play a major role indetermining the environmental lifetime of HBCD since, upon release into the environment, the majority of HBCD is likely to partition into the soil or sediment compartments. In this study, HBCD loss was observed in both viable and abiotic sediments although the rates were appreciable faster inthe viable sediments. Biologically mediated transformation processes (i.e., biotransformation) accelerated the rate of loss of HBCD when compared to the biologically inhibited (i.e., heat-treated) microcosms. Brominated degradation products were not detected inany ofthe sediment microcosms.
Over the last several years a number of international protocols (EC 1996, UNECE 1998, UNEP 2000) have been put forth for the classification of chemicals as persistent (P), bioaccumulative (B), and toxic (T). The criteria for persistence inthese initiatives includes half-lives insoil and sediments ranging from 120 to 180 days. In this investigation the resulting biotransformation half-lives for HBCD inthe two river systems were determined to be 11 and 32 days inthe aerobic sediments and 1.l and 1.5 days inthe anaerobic sediments, respectively.
Limited information is available for the reactions of HBCD inthe 27188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

environment. However, the aerobic degradation and mineralization of a similar type of cyclic aliphatic halogenated fire retardant, FR-651A (mixture of pentabromo-chlorocyclohexane, tetrabromo-dichlorocyclohexane, and tribromotrichlorocyclohexane) has been observed. A soil half-life of -11 days, based upon disappearance of 14C-FR-651A from soil, was reported. Complete degradation of 14C-FR-651A was also observed with a mineralization half-life on the order of 93 days.
An examination of the reactions of brominated aliphatic compounds that contain structural features similar to HBCD can also provide insight into the possible reaction pathways available for HBCD. Brominated aliphatic compounds are known to be susceptible to both hydrolytic and nuclelophilic attack. The reactivity of halogenated aliphatic compounds depends of the strength ofthe carbon-halogen bond, and increases inthe order of F < Cl < Br. For example, the hydrolysis half-lives at pH 7 and 25 “C for methyl fluoride, methyl chloride, and methyl bromide are 1.l x 104, 340, and 20 days, respectively. At environmental pH’s neutral and base catalyzed hydrolysis reactions are most important, and depend on the degree and patterns of halogen substitution. Sulfur based nucleophiles can react rapidly with halogenated aliphatic compounds, thereby reducing their lifetimes inthe environment. The half-life of 1-bromohexane inwater at 25 “C was reduced from 20 days to approximately one day in5 mM HS- or 0.07 mM polysulfide (S&Z-). Similarly, the half-life of 1,2-dibromoethane in water at 25°C was reduced from 1,000 days to 4days inthe presence of5 mM HS-. Sulfide and polysulfides would be expected to be present in sediments at low redox potentials. Thus, reaction of HBCD with these sulfur-based nucleophiles may partly explain the loss of HBCD inthe microcosm studies. Halogenated aliphatic compounds are susceptible to reductive dehalogenation reactions in natural reducing environments. The rates of reactions vary depending on the strength and electron affinity of the carbon-halogen bond, and the stability ofthe carbon-radical species resulting from the initial electron transfer that occurs. Vicinal dehalogenation reactions are particularly important, and are dependent on the halogen atom. Half-lives for vicinal dehalogenation of 1,2-diiodoethane, 1,Bdibromoethane, and 1,2-dichloroethane are 0.6, 55, and 1.8 x 104 hours, respectively. The rapid disappearance of HBCD inthe anaerobic sediment microcosms may be partly explained by reductive dehalogenation reactions. In addition, the disappearance of HBCD inthe aerobic sediment microcosms may also be at least partly explained by reductive dehalogenation reactions. Anaerobic gradients often occur below the surface of sediments that are exposed to an aerobic water column. Such gradients would be expected to form inthe static microcosms used inthis study.
Based upon these results, HBCD is not persistent. Its half-lives in sediment are below the criteria specified by the various international protocols and specifically below the 120 days value specified inthe European Commission’s Technical Guidance Document on Risk Assessment (EC 1996).

Davis J, Gonsior S and Marty G. Evaluation Of Aerobic And Anaerobic Transformation Of Hexabromocyclododecane InAquatic Sediment Systems. Study Number 021081. Environmental Chemistry Research Laboratory, Toxicology & Environmental Research and Consulting. The Dow Chemical Company Midland, Michigan. 2003. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 12.01.2005 (8)
28188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005


### Type lnoculum Contact time Degradation Result Deg. product Method Year GLP Test substance
Attached document anaerobic other: water/sediment microcosms 119 day(s)

= 100

- (k) % after 7 

### day(s) other: half-lives in2 river sediments = 1.l and 1.5 days inthe anaerobic microcosms. not measured other: OECD 308 2003 yes other TS

: Transformation inAerobic 

### andAnaerobic Water/Sediment Microcosms (Sponsor: ACC Brominated Flame Retardant Industry Panel and Dow Chemical Company)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. Its composition was 8.68% alpha isomer, 6.12% beta isomer and 85.19% gamma isomer. This study was performed according to Good Laboratory Practices and OECD Guideline 308.
The transformation of hexabromocyclododecane (HBCD) was determined inaerobic and anaerobic water/sediment microcosms based on the Organization for Economic Co-Operation and Development (OECD) Test Guideline 308 “Aerobic and Anaerobic Transformation inAquatic Sediment Systems.” Laboratory batch microcosms were prepared with authentic water and sediment collected from two rivers inthe eastern United States. Aerobic microcosms were pre-incubated at 20 f 1“C for 49 days and maintained by periodically exchanging the headspace of the microcosms with ambient air to replenish oxygen. Anaerobic microcosms were prepared inan anoxic atmosphere (70% N2,28% C02, and 2% H2). The microcosms were preincubated at23 f 1“C for 43 to 44 days to allow the microcosms to stabilize. HBCD was then added to the microcosms at nominal concentrations ranging from 34 to 89 rig/g (sediment dry weight). Biologically inhibited (i.e., abiotic) controls were prepared by steam sterilization ofthe sediment/water mixture prior to the addition of HBCD. Microcosms were incubated inthe dark at 20 f 1“C for 119 days. The concentration of HBCD inthe microcosms was determined at selected time intervals inthe water and sediment phases utilizing high performance liquid chromatography-mass spectrometry (LC-MS). Aerobic microcosms were analyzed on days 0, 1,7,21,64,91, and 119, while anaerobic microcosms were analyzed on days 0, 1,7, 14, 61(or 62), 91, and 119. HBCD concentrations decreased over time in both the aerobic and anaerobic microcosms. HBCD concentrations inthe viable aerobic microcosms from both river systems decreased at least 90% within 21 days, while the corresponding decreases inthe abiotic controls ranged from 7to 62%. Disappearance of HBCD was observed in both the viable and abiotic anaerobic microcosms with the rate of loss more rapid inthe viable microcosms, with HBCD reaching non-detected levels within 7 days. In contrast, HBCD concentrations inthe abiotic controls decreased from 48 to 62% after 14 days. Pseudo-first order kinetics rate constants for the biotransformation of HBCD were determined by subtracting the abiotic rate constant from the viable rate constant. Biotransformation half-lives for HBCD inthe two river systems were determined to be 11 and 32 days inthe aerobic microcosms and 1.l and 1.5 days inthe anaerobic microcosms. Brominated degradation products were not detected inthe sediment and water layers 29188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

or inthe headspaceofthe microcosms.
The purposeofthisstudywasto determinethe environmental lifetimeof HBCD underrealisticenvironmental conditions. Laboratory microcosms were usedto evaluatethetransformation of HBCD inaerobicand anaerobicwater/sediment microcosms.Sediment degradation processesareexpected to play amajorroleindeterminingthe environmental lifetimeof HBCDsince,upon releaseintothe environment, the majorityof HBCD islikelyto partition intothe soilorsediment compartments. Inthisstudy, HBCD losswasobserved inbothviable and abioticsedimentsalthoughthe rateswere appreciablefaster intheviable sediments.Biologically mediatedtransformation processes(i.e., biotransformation)acceleratedthe rate of lossof HBCDwhen comparedto the biologicallyinhibited(i.e., heat-treated) microcosms.Brominated degradationproductswere notdetected inanyofthesediment microcosms. Overthe lastseveralyears anumberof internationalprotocols(EC 1996, UNECE 1998,UNEP2000)have been putforth forthe classificationof chemicalsaspersistent(P), bioaccumulative (B), andtoxic (T).The criteria for persistenceintheseinitiatives includeshalf-lives insoilandsediments rangingfrom 120to 180days. Inthisinvestigationthe resulting biotransformationhalf-livesfor HBCD inthetwo river systemswere determinedto be 11and32days inthe aerobicsedimentsand 1.l and 1.5 days inthe anaerobicsediments,respectively.
Limitedinformationisavailablefor the reactionsof HBCDinthe environment. However, the aerobicdegradation andmineralizationof a similartype ofcyclic aliphatichalogenatedfire retardant, FR-651A(mixture of pentabromo-chlorocyclohexane, tetrabromo-dichlorocyclohexane, and tribromotrichlorocyclohexane) has beenobserved. Asoilhalf-lifeof-11 days, basedupondisappearanceof 14C-FR-651 Afromsoil,was reported. Complete degradationof 14C-FR-651A wasalsoobservedwith a mineralizationhalf-life onthe orderof 93days.
An examinationofthe reactionsof brominatedaliphaticcompoundsthat contain structural featuressimilarto HBCD can alsoprovide insightintothe possiblereactionpathways availablefor HBCD. Brominatedaliphatic compoundsare knownto besusceptibleto both hydrolytic and nuclelophilic attack. The reactivity of halogenatedaliphaticcompoundsdependsofthe strength ofthe carbon-halogen bond,and increasesinthe orderof F< Clc Br. Forexample,the hydrolysis half-lives at pH 7and25 “Cfor methyl fluoride, methyl chloride, andmethyl bromideare 1.l x 104,340,and20 days, respectively. At environmental pH’sneutral and basecatalyzed hydrolysis reactionsare mostimportant, anddependonthe degree and patterns of halogensubstitution. Sulfur basednucleophilescan react rapidlywith halogenatedaliphaticcompounds,thereby reducingtheir lifetimesinthe environment. The half-lifeof 1-bromohexaneinwater at25 “Cwas reducedfrom20daysto approximately onedayin5 mM HS- or 0.07 mM polysulfide(Sx2-). Similarly,the half-life of 1,Bdibromoethane in water at25°Cwas reducedfrom 1,000daysto4 days inthe presenceof5 mM HS-. Sulfideand polysulfideswould be expected to bepresent in sedimentsat low redox potentials.Thus, reactionof HBCDwiththese sulfur-basednucleophilesmaypartly explainthe lossof HBCDinthe microcosmstudies. Halogenatedaliphaticcompoundsaresusceptibleto reductive dehalogenation reactionsinnatural reducingenvironments.The ratesof reactionsvary dependingonthestrength andelectron affinity ofthe carbon-halogen bond, andthe stability ofthe carbon-radicalspecies resultingfromthe initialelectrontransfer thatoccurs. Vicinal dehalogenation reactionsare particularly important, andare dependenton 30188

## 3. Environmental

Reliability Flag 12.01.2005 Type lnoculum Contact time Degradation Result Deg. product Method Year GLP Test substance
Attached document 

## Fate and Pathways Id 25637-99-4 Date 31.03.2005

the halogen atom. Half-lives for vicinal dehalogenation of 1,2-diiodoethane, 1,Bdibromoethane, and 1,2-dichloroethane are 0.6, 55, and 1.8 x 104 hours, respectively. The rapid disappearance of HBCD inthe anaerobic sediment microcosms may be partly explained by reductive dehalogenation reactions. Inaddition, the disappearance of HBCD inthe aerobic sediment microcosms may also be at least partly explained by reductive dehalogenation reactions. Anaerobic gradients often occur below the surface of sediments that are exposed to an aerobic water column. Such gradients would be expected to form inthe static microcosms used inthis study.
Based upon these results, HBCD is not persistent. Its half-lives in sediment are below the criteria specified by the various international protocols and specifically below the 120 days value specified inthe European Commission’s Technical Guidance Document on Risk Assessment (EC 1996).

Davis J, Gonsior S and Marty G. Evaluation Of Aerobic And Anaerobic Transformation Of Hexabromocyclododecane In Aquatic Sediment Systems. Study Number 021081. Environmental Chemistry Research Laboratory, Toxicology & Environmental Research and Consulting. The Dow Chemical Company Midland, Michigan. 2003. : (1) valid without restriction

.

. Critical study for SIDS endpoint (8)

.

. aerobic

. . other: soil microcosms

.

. 119 day(s) : 

### = 75

- (3 % after 119 day(s)

.

. other: half-life

- 63 days

.

. not measured

. . other: OECD 308 : 

### 2003 : 

### yes : as prescribed by 1.l

- 1.4

: Transformation inAerobic and Anaerobic Soil Microcosms (BFRIP and Dow Chemical Company)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This 

### study was performed according to Good Laboratory Practices.
The transformation of HBCD was determined inaerobic and anaerobic soils based on the Organization for Economic Co-Operation and Development (OECD) Test Guideline 307 ‘Aerobic and Anaerobic Transformation in Soil.” Soil microcosms were prepared by adding asandy loam surface soil to serum bottles sealed with Teflon C coated septa. Aerobic microcosms were prepared by adjusting the soil moisture to 20% (by weight) and periodically exchanging the headspace of the microcosms with ambient air to replenish oxygen. The microcosms were pm-incubated at 20 f 1“C for 35 days. Anaer0bi.c microcosms were prepared inan anaerobic atmosphere (70% N2,28% C02, and 2% H2) by flooding the soil with water and pre-incubating the microcosms at 23 f 1“C for 43 days to allow low redox (e.g., methanogenic) conditions to develop. HBCD was then added to microcosms at a nominal concentration of 25 rig/g (soil dry weight), together with activated sludge (5 mg/g, dry weight basis) from a municipal wastewater treatment plant to simulate sludge land treatment applications. Biologically inhibited (abiotic) controls were prepared by 31 I88

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

steam sterilization prior to the addition of HBCD. Microcosms were incubated inthe dark at 20 f 1“C for 119 days. The concentration of HBCD inthe microcosms was determined at selected time intervals utilizing high performance liquid chromatography-mass spectrometry (LCMS). Aerobic microcosms were analyzed on days 0, 1,7,21,48,65, and 119, while anaerobic microcosms were analyzed on days 0, 1,7, 21, 56, 91, and 119. HBCD concentrations decreased over time in both the aerobic and anaerobic soils. HBCD concentrations decreased 75% over 119 days inthe viable aerobic soil microcosms, compared to a 3% decrease inthe abiotic controls, indicating that biological processes were responsible for most of the losses observed. Under anaerobic conditions, HBCD concentrations decreased 92% over 21 days inthe viable microcosms compared to a less than 1% decrease inthe abiotic controls. Pseudo-first order kinetics rate constants for the biotransformation of HBCD were determined by subtracting the abiotic rate constant from the viable rate constant. Biotransformation half-lives for HBCD were determined to be 63 and 6.9 days inthe aerobic and anaerobic soils, respectively. Brominated degradation products were not detected inthe soil or inthe headspace of the microcosms.
The purpose of this study was to determine the environmental lifetime of HBCD under realistic environmental conditions. Laboratory microcosms were used to evaluate the transformation of HBCD inaerobic and anaerobic soils. Soil degradation processes are expected to play a major role indetermining the environmental lifetime of HBCD since, upon release into the environment, the majority of HBCD is likely to partition into the soil or sediment compartments. Inthis study, HBCD loss was observed in both viable and abiotic soil microcosms although the rates were appreciably faster inthe viable soils. Biologically mediated transformation processes (i.e. biotransformation) accelerated the reate of loss of HBCD when compared to the biologically inhibited (ie. heat-treated) soils. No bromianted degradaiton products were observed in either system. Over the last several years a number of international protocols (EC 1996, UNECE 1998, UNEP 2000) have been put forth for the classification of chemicals as persistent (P), bioaccumulative (B), and toxic (T). The criteria for persistence inthese initiatives includes half-lives insoil and sediments ranging from 120 to 180 days. Inthis investigation the resulting biotransformation half-lives for HBCD were determined to be 63 and 6.9 days inthe aerobic and anaerobic soils, respectively. Based upon these results HBCD is not persistent. The half-lives insoil are below the criteria specified by the various international protocols.
Limited information is available for the reactions of HBCD inthe environment. However, the aerobic degradation and mineralization ofa similar type of cyclic, aliphatic halogenated fire retardant, FR-651 A (mixture of pentabromochlorocyclohexane; tetrabromo-dichlorocyclohexane, and tribromotrichlorocyclohexane) was observed. A soil half-life of

- 11days based upon disappearance of 14 C-FR-651 A from soil was reported. Complete degradation of 14C-FR-651 A was also observed with mineralization half-life on the order of 93 days.
An examination of the reactions of brominated aliphatic compounds that contain structural features similar to HBCD can also provide insight into the possible reaction pathways available for HBCD. Brominated aliphatic compounds are known to besusceptible to both hydrolytic and nuclelophilic attack. The reactivity of halogenated aliphatic compounds increases inthe order of F < Cl < Br. For example, the hydrolysis half-lives at pH 7and 25 “C for methyl fluoride, methyl chloride, and methyl bromide are 1.l x 104, 340, and 20 days, respectively. At environmental pH’s, neutral and base 32188

## 3. Environmental Fate and Pathways Id Date 25637-99-4 31.03.2005

- catalyzed hydrolysis degree and patterns reactions are most important, of halogen substitution. and depend on the

Sulfur based nucleophiles can react rapidly with halogenated aliphatic compounds, thereby reducing their lifetimes inthe environment. The halflifeof 1-bromohexane inwater at 25 “C was reduced from 20 days to approximately one day in5 mM HS

- or 0.07 mM polysulfide (Sx 2- ). Similarly, the half-life of 1,2-dibromoethane in water at 25 “C was reduced from 1,000 days to 4days inthe presence of 5 mM HS-. Sulfide and polysulfides would be expected to be present insoils at low redox potentials. Thus, reaction of HBCD with these sulfur-based nucleophiles may partly explain the loss of HBCD inthe soil studies. Since HBCD contains three pairs of vicinal bromine atoms, the transformation of simple aliphatic compounds containing vicinal bromine atoms may provide insight into possible reaction pathways for HBCD. For example, 1,2-dibromoethane reacts with nucleophiles via both substitution and elimination reactions. Reaction with HS- via an “SN2” substitution reaction reaction results inthe formation of HS-CHBCH2SH, while an elimination under alkaline conditions results inthe formation of H2C=CHBr. A combination of elimination and substitution reactions can result inthe formation of a mixture of HO-H2-CH2-OH and H2C=CHBr. Similar mechanisms study. may be responsible for the loss of HBCD observed inthis
Halogenated aliphatic compounds are susceptible to reductive dehalogenation reactions in natural reducing environments. The rates of reactions vary depending on the strength and electron affinity of the carbon-halogen bond and the stability of the carbon-radical species resulting from the initial electron transfer that occurs. Vicinal dehalogenation reactions are particularly important, and are dependent on the halogen atom. Half-lives for vicinal dehalogenation of 1,2-diiodoethane, 1,2-dibromoethane, and 1,Bdichloroethane are 0.6, 55, and 1.8 x 104 hours, respectively. The rapid disappearance of HBCD inthe anaerobic soil microcosms may be partly explained by reductive dehalogenation reactions.
Based upon these results, HBCD is not persistent. clearly below the criteria for persistence specified protocols (UNECE 1966, UNEP 2001). Its half-lives insoil are invarious international

Reliability Flag 12.01.2005 : : Davis J, Gonsior S and Marty G. 2003. Evaluation Of Aerobic And Anaerobic Transformation Of Hexabromocyclododecane In Soil. Study Number 021082. Environmental Chemistry Research Laboratory. Toxicology & Environmental Research and Consulting. The Dow Chemical Company. Midland, Ml. (1) valid without restriction Critical study for SIDS endpoint (5) 

## Type lnoculum . . : anaerobic other: soil microcosms Contact time : 119 day(s) Degradation : = 92

- (3 % after 21 day(s) Result : other: half-life -6.9 days Deg. product : not measured Methbd : other: OECD 307 Year : 2003 GLP Test substance : : yes as prescribed by 1.1

- 1.4 Attached document : Transformation inAerobic and Anaerobic Soil Microcosms (Sponosr: ACC 33188

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

BFRIP andDowChemicalCompany)
Thetest articlewasa compositeof3lotsofcommercialHBCD produced byAlbemarleCorporation, Great LakesChemicalCorporation, and Ameribrom, Inc. Thisstudywasperformedaccordingto Good Laboratory Practices.
Thetransformation of HBCDwas determinedinaerobicandanaerobic soilsbasedonthe Organization for EconomicCo-Operation and Development (OECD)Test Guideline307“Aerobic andAnaerobic Transformation in Soil.” Soil microcosmswere preparedby addingasandy loamsurface soiltoserumbottlessealedwithTeflon C coated septa. Aerobic microcosmswere prepared byadjustingthe soilmoistureto20% (byweight) and periodicallyexchangingthe headspaceofthe microcosms with ambientairto replenishoxygen. The microcosmswere preincubated at20f 1“Cfor 35 days.Anaerobic microcosmswere prepared inan anaerobicatmosphere(70% N2,28%C02, and2% H2)byfloodingthe soilwithwaterand pre-incubatingthe microcosmsat23f 1“Cfor43 days to allowlow redox (e.g., methanogenic)conditionsto develop. HBCDwas then addedto microcosmsata nominalconcentration of25 rig/g (soildry weight),together with activated sludge(5 mg/g,dryweight basis)from a municipalwastewater treatment planttosimulatesludgelandtreatment applications. Biologicallyinhibited(abiotic) controlswere prepared by steamsterilization priorto the additionof HBCD. Microcosmswere incubated inthe darkat20f 1“Cfor 119days.The concentration of HBCDinthe microcosmswasdeterminedatselectedtimeintervals utilizing highperformance liquidchromatography-massspectrometry (LCMS). Aerobic microcosmswere analyzed on days 0, 1,7,21,48, 65, and 119,whileanaerobic microcosmswere analyzed ondays 0, 1,7,21, 56, 91, and 119. HBCDconcentrations decreased.over time inboththe aerobicand anaerobicsoils.HBCD concentrations decreased75%over 119days inthe viable aerobicsoilmicrocosms,comparedto a3% decreaseinthe abiotic controls, indicatingthat biologicalprocesseswere responsiblefor mostof the lossesobserved. Underanaerobicconditions, HBCDconcentrations decreased92% over 21days intheviable microcosmscomparedto aless than 1%decreaseinthe abioticcontrols. Pseudo-firstorder kineticsrate constantsfor the biotransformationof HBCDwere determined by subtractingthe abiotic rateconstantfromtheviable rate constant. Biotransformationhalf-livesfor HBCDwere determinedto be63 and6.9 daysinthe aerobicandanaerobicsoils,respectively. Brominated degradation productswere notdetected inthesoilor inthe headspaceof the microcosms.
The purposeofthisstudywasto determinethe environmental lifetimeof HBCDunder realisticenvironmental conditions. Laboratory microcosms were usedto evaluate thetransformation of HBCD inaerobicand anaerobicsoils.Soildegradation processesare expected to playamajor roleindeterminingthe environmental lifetimeof HBCDsince, uponrelease intothe environment, the majorityof HBCDislikelyto partition intothesoil orsedimentcompartments. Inthisstudy, HBCDlosswasobserved inboth viable and abioticsoilmicrocosmsalthoughthe rateswere appreciably faster intheviable soils. Biologicallymediatedtransformation processes (i.e., biotransformation)acceleratedthe rate of lossof HBCDwhen comparedtothe biologically inhibited(i.e., heat-treated) soils.No brominateddegradation productswere observed ineithersystem. Over the lastseveralyears anumberof international protocols (EC 1996, UNECE 1998,UNEP2000) have been putforth forthe classificationof chemicalsaspersistent (P), bioaccumulative (B), andtoxic (T).The criteria for persistenceinthese initiatives includeshalf-lives insoilandsediments

. 

## was

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

ranging from 120 to 180 days. Inthis investigation the resulting biotransformation half-lives for HBCD were determined to be 63 and 6.9 days inthe aerobic and anaerobic soils, respectively. Based upon these results HBCD is not persistent. The half-lives insoil are below the criteria specified by the various international protocols.
Limited information is available for the reactions of HBCD inthe environment. However, the aerobic degradation and mineralization of a similar type of cyclic, aliphatic halogenated fire retardant, FR-651A (mixture of pentabromochlorocyclohexane, tetrabromo-dichlorocyclohexane, and tribromotrichlorocyclohexane) was observed. A soil half-life of -11 days based upon disappearance of 14 C-FR-651A from soil was reported. Complete degradation of 14C-FR-651 Awas also observed with mineralization half-life on the order of 93 days.
An examination of the reactions of brominated aliphatic compounds that contain structural features similar to HBCD can also provide insight into the possible reaction pathways available for HBCD. Brominated aliphatic compounds are known to be susceptible to both hydrolytic and nuclelophilic attack. The reactivity of halogenated aliphatic compounds increases inthe order of F < Cl < Br. For example, the hydrolysis half-lives at pH 7 and 25 “C for methyl fluoride, methyl chloride, and methyl bromide are 1.1 x 104, 340, and 20 days, respectively. At environmental pH’s, neutral and base catalyzed hydrolysis reactions are most important, and depend on the degree and patterns of halogen substitution.
Sulfur based nucleophiles can react rapidly with halogenated aliphatic compounds, thereby reducing their lifetimes inthe environment. The halflifeof 1-bromohexane inwater at 25 “C was reduced from 20 days to approximately one day in5 mM HS

- or 0.07 mM polysulfide (Sx 2- ). Similarly, the half-life of 1,Bdibromoethane in water at 25 “C was reduced from 1,000 days to 4 days inthe presence of 5 mM HS-. Sulfide and polysulfides would be expected to be present insoils at low redox potentials. Thus, reaction of HBCD with these sulfur-based nucleophiles may partly explain the loss of HBCD inthe soil studies.
Since HBCD contains three pairs of vicinal bromine atoms, the transformation of simple aliphatic compounds containing vicinal bromine atoms may provide insight into possible reaction pathways for HBCD. For example, 1,2-dibromoethane reacts with nucleophiles via both substitution and elimination reactions. Reaction with HS- via an “SN2’ substitution reaction results inthe formation of HS-CH2-CH2-SH, while an elimination reaction under alkaline conditions results inthe formation of H2GCHBr. A combination of elimination and substitution reactions can result inthe formation of a mixture of HO-H2-CH2-OH and H2C=CHBr. Similar mechanisms may be responsible for the loss of HBCD observed inthis study. Halogenated aliphatic compounds are susceptible to reductive dehalogenation reactions in natural reducing environments. The rates of reactions vary depending on the strength and electron affinity of the carbon-halogen bond and the stability of the carbon-radical species resulting from the initial electron transfer that occurs. Vicinal dehalogenation reactions are particularly important, and are dependent on the halogen atom. Half-lives for vicinal dehalogenation of 1,Bdiiodoethane, 1,Bdibromoethane, and 1,Bdichloroethane are 0.6, 55, and 1.8 x 104 hours, respectively. The rapid disappearance of HBCD inthe anaerobic soil microcosms may be partly explained by reductive dehalogenation reactions.
Based upon these results, HBCD is not persistent. Its half-lives insoil are clearly below the criteria for persistence specified invarious international 35 / 88

## 3. Environmental

Reliability Flag 12.01.2005 Type lnoculum Contact time Degradation Result Deg. product Method Year GLP Test substance Deg. products

Attached document 

## Fate and Pathways Id 25637-99-4 Date 31.03.2005

protocols (UNECE 1966, UNEP 2001).
Davis J, Gonsior S and Marty G. 2003. Evaluation Of Aerobic And Anaerobic Transformation Of Hexabromocyclododecane In Soil. Study Number 021082. Environmental Chemistry Research Laboratory. Toxicology & Environmental Research and Consulting. The Dow Chemical Company. Midland, Ml.

.

. (1) valid without restriction

. Critical study for SIDS endpoint (5)

other:activated and digester sludge, river sediment, and surface soil

- (*) % after other: degradation of all isomers inanaerobic digester sludge and in freshwater aerobic and anaerobic sediment microcosms yes other:OECD 3028; 307,308; IS0 11734 2004 yes other TS: [14C]HBCD dibromocyclododecadiene tetrabromocyclododecene 27070-59-3 248-206-l cyclododecatriene

: Investigation of the biodegradation of [14C]Hexabromocyclododeoane in sludge, sediment, and soil (European Brominated Flame Retardant Industry Panel)
This study investigated the biodegradation ofthe three HBCD stereoisomers, alpha, beta and gamma, and the identity of major degradation products. The formation and identification of degradation products of HBCD was assessed inactivated and digester sludge, river sediment, and surface soil. Both aerobic and anaerobic biodegradation were evaluated in laboratory reaction mixtures and batch microcosms. This study was performed according to the relevant OECD Guidelines (302B; 307,308), IS0 11734, and Good Laboratory Practices.
To generate sufficient levels of [14C]degradation products for their identification, [14C]HBCD was added to reaction mixtures and microcosms at nominal concentrafions ranging from 3to 5 mg/kg (or mg/L), exceeding the water solubility ofthe test material by greater than an order or magnitude. Duration of the studies ranged from approximately 60 to 112 days. Reaction mixtures and batch microcosms were extracted and analyzed by high performance liquid chromatography (HPLC) with radiochemical detection to follow the degradation of the 3stereoisomers and formation of [14C]products. Product identification was facilitated by analyses of extracts from the soil, sediment and sludge mixtures by HPLCatmospheric pressure photo ionization-mass spectrometry (APPI-MS) or gas chromatography-electron impact ionization-mass spectrometry (GC-EIMS). Substantial biological transformation of [14C]HBCD was observed inthe anaerobic digester sludge and infreshwater aerobic and anaerobic sediment microcosms. Conversely, no degradation of HBCD was noted in the soil microcosms incubated under aerobic conditions. Inthe digester sludge and sediment, degradation of each of the three stereoisomers occurred over the course of the study. Little difference was noted inthe disappearance ofthe three stereoisomers, indicating similarity inthe extent of degradation of each isomer.
36 I88

## 3. Environmental Fate and Pathways Id 25637-99-4 Date 31.03.2005

Concomitant with the loss of [14C]HBCD inthe sludge and sediment test mixtures was formation ofthree [14C]products. Using a combination of HPLC-APPI-MS and GC-El-MS these metabolites were identified as tetrabromocyclododecene, dibromocyclododecadiene, and cyclododecatriene. These products suggest HBCD is sequentially debrominated via dihaloelimination by naturally occurring microorganisms inwastewater sludge and aquatic sediment. During each sequential debromination, two bromines are lost from vicinal carbons with the subsequent formation of a double bond between the.adjacent carbon atoms. These results demonstrate microorganisms naturally occurring in aquatic sediment and wastewater sludges can completely debrominate HBCD.
Davis JW, Gonsior SJ, Markham DA, and Marty GT. 2004. Investigation of the biodegradation of [14C]hexabromocyclododecane insludge, sediment, and soil. Laboratory Project Study ID 031178. Toxicology & Environmental Research and Consulting. The Dow Chemical Company, Midland, Ml. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 11.01.2005 (9)

3.6 BOD5, COD OR BODYCOD RATIO

3.7 BlOACCUMULAlION

Species : Oncorhynchus mykiss (Fish, fresh water) Exposure period : 34 day(s) at 25 “C Concentration : 3.4jlgll BCF .

. = 8974

- Elimination : yes Method : EPA OPPTS 850.1730 Year : 2000 GLP : yes Test substance : as prescribed by 1.l

- 1.4
Attached document : Flow Through Bioconcentration In Rainbow Trout (Oncorhynchus mykiss) (Sponsor: ACC BFRIP)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarfe Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD and GLP guidelines.
Nominal test concentrations were 0, 0.34, and 3.4 ug HBCD/L. These doses are equivalent to gamma HBCD’s water solubility and one tenth of its water solubility. Mean measured (LCIMS with heated nebulizer operated in the selected ion monitoring mode) test concentrations were 0, 0.18, and 1.8 ug HBCD/L. The length ofthe test was 70 days (35-day uptake, 35-day depuration). The steady bioconcentration factor (BCF) at a nominal concentration of 3.4 ug HBCD/L (1.8 ug HBCD/L measured) inwhole fish was 8,974. This BCF was further defined as 4,850 in edible tissue and 12,866 in non-edible tissue. Steady state was not achieved at the nominal concentration of 0.34 ug HBCD/L due to an unexpected increase intissue concentrations at day 35. The unexpected increase intissue concentrations on day 35 may have been due tothe variability inthe measured water concentrations inthis treatment group. The variability in turn is likely afunction of the extremely low nominal concentration atthis
37188

## 3. Environmental

Reliability Flag 28.12.2004 Species Exposure period Concentration BCF Elimination Method Year GLP Test substance
Attached document

Reliability Flag 29.12.2004 

## Fate and Pathways Id 25637-99-4 Date 31.03.2005

dose level (0.34 ug HBCD/L). Thus, the calculated BCF inthe nominal 3.4 ug HBCD/L treatment group is considered a better estimate than that inthe 0.34 ug HBCD/L treatment group. Drottar K. and Krueger H. 2000. Hexabromocyclododecane (HBCD): Flowthrough bioconcentration test with rainbow trout (Oncorhynchus mykiss). Project No.: 439A-111. Wildlife International, Ltd. Easton, MD. : (1) valid without restriction

. . Critical study for SIDS endpoint / (10)

: other: Eisenia fetida : 28 day(s) at “C : : = 4.5

- : :

.

. 2002 : yes : as prescribed by 1.1

- 1.4

.

. Earthworm (Eisenia fetida) Survival and Reproduction
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD and GLP guidelines.
The artificial soil used inthis study was characterized as sandy loam with an 80% sand, 8% silt, and 12% clay content. Nominal test concentrations were 0,78.5, 157, 313, 625, 1,250,2,500 and 5,000 mg HBCD/kg of dry soil. Mean measured concentrations at day 28 were cl .28 (Control), 61.2, 145, 244, 578, 1,150, 2,180, and 4,190 mg HBCD/kg of dry soil. Mean measured concentrations at day 56 were 56 Days: cl.35 (Control), 51.5, 128,235, 543, 1,070, 2,020, and 3,990 mg HBCD/kg of dry soil. Measured tissue concentrations at day 28 were <0.200 (control), 3.40, 7.32, 16.8, 15.3, 53.0, 71.2, and 150 mg HBCD per gram of tissue.
No abnormal burrowing or avoidance behaviors were recorded during the first 60 minutes of testing. Using the estimated soil Koc and the formula provided inthe European Union’s Technical Guidance Document, HBCD’s calculated BCF in earthworms is:
BCFearthworm

= Cearthworm / Csoil

= Kearthworm-porewater x RHOsoil x 10+3 / Ksoil-water =(0.15 mg/kg)(1.25 x 10+5)/4,190 mg/kg dry soil

= 4.5. Thus, HBCD did not bioconcentrate inthe earthworm.

Aufterheide et al. 2002. Effect of Hexabromocyclododecane on the Survival and Reproduction of the Earthworm, Eisenia fetida. ABC Study No. 47222. ABC Laboratories, Inc., Columbia, Missouri; Wildlife International, Inc., Easton, MD. (1) valid without restriction Critical study for SIDS endpoint (11) 3818827.12.2004 

## 3. Environmental Fate and Pathways id 25637-99-4 Date 31.03.2005

3.8 ADDITIONAL REMARKS

Memo : Summary: HBCD Half-lives in Sediment and Soil
Attached document : Substances which are persistent is soil or sediments are typically defined as those wiht half-lives ranging from 120 to 180 days. HBCD’s half-lives in aerobic and anaerobic soils were 63 and 6.9 days, respectively. In sediments collected from two river systems, HBCD’s half-lives were 11and 32 days inthe aerobic water/sediment microcosms and 1.1 and 1.5 days in the anaerobic water/sediment microcosms.
Based upon these results, HBCD is not persistent. Its half-lives insoil and sediment are below the criteria specified by various international protocols (UNECE 1966, UNEP 2001, EU Technical Guidance Document 1996). 12.01.2005

39188

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

4.1 ACUTE/PROLONGED TOXICITY TO FISH


### Type : flow through Species : Oncorhynchus mykiss (Fish, fresh water) Exposure period : 96 hour(s) Unit : Pug/l NOEC : >= 6.8

- measured/nominal Limit test : no Analytical monitoring : yes Method : EPA OPPTS 850.1075 Year : 1997 GLP : yes Test substance : as prescribed by 1.1

- 1.4
Attached document : 96-Hour Acute Toxicity Test With Rainbow Trout (Oncorhynchus mykiss) (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD guidelines and Good Laboratory Practices.
HBCD was not acutely toxic to rainbow trout at the limitof the gamma stereoisomer’s solubility. HBCD’s 96 hour LC50, no mortality concentration and no observed effect concentration were all> than the gamma stereoisomer’s water solubility. The highest nominal dose tested was twice that water solubility. Nominal test concentrations were 0, 1.5, 2.2, 3.2, 4.6 and 6.8 ug/L and corresponded to mean measured concentrations (HPLC with UVNIS detector) of 0, 0.75, 1.5, 2.3, 2.3 and 2.5 q/L, respectively. Graves, W and Swigert, J. (1997) Hexabromocyclododecane (HBCD): A 96-Hour Flow-Through Acute Toxicity Test with the Rainbow Trout (Oncorhynchus mykiss). Project Number: 439A-101. Wildlife International LTD, Easton, MD. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 29.12.2004 (12) Type : flow through Species : Oncorhynchus mykiss (Fish, fresh water) Exposure period : 88 day(s) Unit : iJs/l NOEC

. . = 6.8

- measured/nominal Limit test : no Analytical monitoring : yes Method : OECD Guide-line 210 Year *

. 2001 GLP : yes Test substance : as prescribed by 1.l

- 1.4
Attached document : Fish Early Life Stage In Rainbow Trout (Oncorhynchus mykiss) (BFRIP)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current OECD guidelines and Good Laboratory Practices. Rainbow trout embryos were exposed to nominal HBCD water concentrations of 0.43, 0.85, 1.7, 3.4 and 6.8 ug/L. The top two doses 40/88

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

represent gamma HBCD’s water solubility (3.4 ug/L) and two times gamma HBCD’s water solubility (6.8 IQ/L). A negative control and solvent control group were also included. Mean measured concentrations (LC/MS with heated nebulizer operated inthe selective ion monitoring mode) were 0.25, 0.47, 0.83, 1.8 and 3.7 ug/L. This method was designed to monitor for all3 HBCD diastereomers; however, the trace residues of the alpha and beta diastereomers were evident inthe water samples were below the established limits of quantitation. Comparison of the chromatograms from study initiation through study termination showed that the relative distribution of the HBCD diastereomers remained constant during the definitive study, and the gamma diastereomer measured results were consistent throughout the study. Hatching success, time to hatch, time for larvae to swim-up, and post-hatch growth and survival were evaluated during the 88-day test. Rainbow trout exposed to HBCD at mean measured concentrations upto 3.7 ug/L (nominal concentration = 6.8 ug/L or twice HBCD’s water solubility) for a 27-day hatching period and 61 days post-hatch showed no effects on hatching success, time to swim-up, larval survival, fry survival or growth. Consequently, HBCD was not chronically toxic to rainbow trout at concentrations at or above its limit ofsolubility. The NOEC for this study was 3.7 ug/L or 6.8 ug/L nominal (twice gamma HBCD’s water solubility). The low-effect-concentration (LOEC) and maximum acceptable toxicant concentration (MATC) could not be determined due to absence oftoxicity, but were considered >3.7 ug/L or >6.8 ug/L nominal (> twice gamma HBCD’s water solubility). Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 28.12.2004 (13)

4.2 ACUTE TOXICITY TO AQUATlC INVERTEBRATES



### Type : flow through Species

. . Daphnia magna (Crustacea) Exposure period

. . 48 hour(s) Unit

. . Pug/l NOEC

. . >= 3.2

- measured/nominal EC50 . > 3.2

- measured/nominal Limit Test . no Analytical monitoring . yes Method : EPA OPPTS 850.1010 Year : 1997 GLP : Yes Test substance : as prescribed by 1.l

- 1.4
Attached document : Acute Toxicity to Aquatic Invertebrates: 48-Hour Acute Toxicity Test With Daphnia magna (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Amerfbrom, Inc. This study was performed according to current EPA, OECD guidelines and Good Laboratory Practices. HBCD was not acutely toxic to Daphnia magna. HBCD’s 48 hour EC50, no mortality/immobility concentration, and no observed effect concentration (6.8 ug/L nominal) in Daphnia magna were all > than gamma HBCD’s water solubility (3.4 ug/L measured). The highest nominal dose tested was twice gamma HBCD’s measured water solubility. Nominal test concentrations were 0, 1.5, 2.2, 3.2, 4.6 and 6.8 ug/L which corresponded 41 I88

## 4. Ecotoxicity

Reliability : Flag : 29.12.2004

4.3 TOXICfTY TO AQUATlC

Species : Endpoint : Exposure period : Unit

. - NOEC : LOEC : EC10 : EC50 : Limit test : Analytical monitoring : Method : Year : GLP : Test substance : Attached document :

Reliability : Flag : 29.12.2004 Species : Endpoint : Exposure period : Unit : NOEC : EC50

. - Limit test : Id 25637-99-4 Date 31.03.2005

to mean measured concentrations (HPLC with UVNIS detector) of 0, 2.4, 1.8, 2.1, 2.3 and 3.2 ug/L, respectively. Graves W and Swigert J. (1997) Hexabromocyclododecane (HBCD): a48- hour flow-through acute toxicity test with the cladoceren (Daphnia magna). Project Number: 439A-102. Wildlife International Ltd., Easton, MD. (1) valid without restriction Risk Assessment, Critical study for SIDS endpoint (14)

PLANTS E.G. ALGAE

Selenastrum caprfcornutum (Algae) biomass 96 hour(s) Pvg/l >= 3.7

- measured/nominal > 3.7

- measured/nominal > 3.7

- measured/nominal > 3.7

- measured/nominal no yes EPA OPPTS 850.5400 1997 yes as prescribed by 1.l

- 1.4
96-Hour Acute Toxicity Test With The Freshwater Alga (Selenastrum capricomutum) (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarfe Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD guidelines and Good Laboratory Practices. This study was performed to complete the EU base set. HBCD was not acutely toxic to Selenastrum capricomutum. HBCD’s 96 hour EC10, EC50, EC90 and no observed effect concentration were all> than HBCD’s water solubility based on the gamma isomer. The highest nominal dose tested was twice gamma HBCD’s water solubility. Dose levels were 0, 1.5, 2.2, 3.12 4.6 and 6.8 ug/L (nominal). The mean measured concentration (HPLC with UV/VIS detector) at the 6.8 ug/L dose was 3.7 ug/L.
Roberts C. and Swigeti J. Hexabromocyclododecane (HBCD): A96Hour Toxicity Test with the Freashwater Alga (Selenastrum capricomutum). Wildlife International Ltd. Project Number: 439A-103. June 3, 1997. Wildlife International Ltd., Easton, MD. (1) valid without restriction Risk Assessment, Critical study for SIDS endpoint (15) Skeletonema costatum (Algae) other:cell densities, biomass and growth rate 72 hour(s) Ps/l <41- measured/nominal >41- measured/nominal yes 42188

## 4. Ecotoxicity

Analytical monitoring Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

: yes

. * OECD Guide-line 201 “Algae, Growth Inhibition Test” : 2004 : yes : as prescribed by 1.l

- 1.4

: HBCD: A 72-Hour toxicity test with the marine diatom (Skeletonema costatum)
The objective of this study was to determine the toxicity ofwater-soluble components of hexabromocyclododecane (HBCD) to the marine diatom Skeletonema costatum, using asaltwater algal media passed through a generator column saturated with HBCD.
This study was performed according to Good Laboratory Practices, IS0 10253:1995, OECD 201, and EU Directive 92/69/EEC, Method C.3. The test article was a composite ofthe commercial products of Albemarle Corporation, Dead Sea Bromine Group and Great Lakes Chemical Corporation. Composite = 7.67% alpha, 5.15% beta and 83.04% gamma. The experimental design was developed based on prelminary work. The marine diatom was exposed to a single test concentration (produced via water generator column) of HBCD insalwater algal media, a negative control (also referred to as Control with Column) and a media control (also referred to as Control no Column) for 72 hours. The single test concentration was prepared using algal media passed through a comlumn generator packed with asolid support coate with HBCD to achieve the test concentraiton of HBCD insaltwater algal medium. Measured test concentrations were determined from samples of test medium collected from teh treament and each control group atthe beginning of the test and at test termination.
At test initiation an inoculum of the algal cells was added to each test chamber to achieve a nominal concentratin of approximately 77,000 Skeletonema cells/ml. Samples were collected from each replicate test chmaber at approximately 24-hour intervals during the test to determine cell densities, which were subsequently used to caluculate areas under the growth curve (biomass) and growth rates. Cell densitites, biomass and growth rates were used to calculate percent inhibitoin vlaues relative to the control over the 72-hour exposure period. No-oberved-effectconcentrations (NOEC) were determined at 72 hours through statistical evaluation ofthe cell densities, biomass and growth rates, as well as examination of the concentration-response pattern.
The analytical method for the 3 HBCD stereoisomers insaltwater algal medium was based on methodology developed by Wildlife International LTD. It used an HPLC interfaced with atriple quadrupole LC mass spectromer operated in single-quadrupole selective ion monitoring mode. The LOCI was 0.5 uga.i./L. The Day 0 and 3 arithmetic mean measured test concentrations for Gamma, Beta, Alpha and total HBCD was 1.61, 8.86, 30.5 and 41.Oug a.i./L, respectively. Arithmetic mean measured test concentrations were used to calculate the NOEC values. S. costatum were exposed to asingle concentraion of 41.Oug a.i./L of toal HBCD and evaluated for effects on cell density, area under the growth curve (biomass) and growth rate. After 72 of exposure, inhibition of cell density, biomass and growth rate inthe treated group was 19,21 and 7.3%, respectively, relative to control no column. Similarly, relative tothe control with column, inhibition of cell density, biomass and growth rate was 31, 31 and 11%, respectively. The treated group was statistically different from both control groups (pcO.05).

43188

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

Since there were effects observed, the 72 hour NOEC and EC50, based on cell density, biomass and growth rate were < 41.OI.@ a.i./L and > 41.Oug a.i./L total HBCD, respectively. The total HBCD concentration was composed of 1.61 ug a.i./L gamma; 8.86 ug a.i./L beta and 30.5 ug a.i./L alpha isomers. Desjardins D, MacGregor J, Krueger H. 2004. Hexabromocyclododecane (HBCD): a72-hour toxicity test with the marine diatom (Skeletonema costatum). Final Report. Wildlife International LTD Project Number: 439A125. Wildlife International LTD, Easton, MD. Reliability : (1) valid without restriction Flag : Risk Assessment, Critical study for SIDS endpoint 29.12.2004 (16) Species : other algae Endpoint : biomass Exposure period : Unit .

. Method : other Year : 1987 GLP : no data lest substance : other TS
Attached document : Walsh et al. 1987 reported testing the effect of media and test chemicals on acute toxicity in marine algae. HBCD was tested in 3species of marine algae in 6different media, and was not toxic at the limits of the gamma isomers water solubility. The EC50’s are as follows:

Chlorella sp 96 hr EC50 > 1500 ug/L); S. costatum 72 hr EC50 9.3-12 ug/L; T. pseudonana 72 hr EC50 50-370 q/L).
Walsh et al. 1987. Ecotoxicology and Environmental Safety, 14, 215-222. Reliability : (2) valid with restrictions 29.12.2004 (17)

4.4 TOXICITY TO MICROORGANISMS E.G. BACTERIA

Type : other Species : activated sludge, domestic Exposure period : 3 hour(s) Unit : mg/l Analytical monitoring : yes Method : OECD Guide-line 209 “Activated Sludge, Respiration Inhibition Test’ Year

. - 2003 GLP : yes Test substance : as prescribed by 1.l

- 1.4

: Activated Sludge Respiration Inhibition Test (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD (209) guidelines and Good Laboratory Practices. HBCD’s effect on the respiration of activated sludge microorganisms was assessed using control, reference and treatment groups. The control group was used to determine the background respiration rate of the sludge and was not dosed with the test or reference substance. The reference group was dosed with 3,5-dichlorophenol, a known inhibitor of respiration, at 44 I88

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

nominal concentrations of 3, 15 and 50 mg/L. The test substance was dosed at a limit concentration of 15 mg/L. After an exposure period of

- three hours, the respiration rates ofthe test solutions were measured using a dissolved oxygen meter. The individual respiration rates of the two controls were 60.5 and 55.5 mg 02Uhr. The difference between the two control respiration rates was 9.0% and was within the 15% difference limit established for the test. The validity ofthe test was further supported by the results from the 3,5-dichlorophenol reference group, which resulted inan EC50 of 5.2 mg/L and was within the 5to 30 mg/L range considered acceptable for the test. An average of 29.1 percent inhibition was observed inthe treatment group. Schaefer E and Siddiqui A. 2003. Hexabromocylcododecane (HBCD): An Activated Sludge, Respiration Inhibition Test. Project Number: 439E-106A. Wildlife International, Ltd. Easton, MD. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 28.12.2004 (6)

4.5.1 CHRONIC TOXIClTY TO .FlSH

Species

. . Oncorhynchus mykiss (Fish, fresh water) Endpoint : other: Hatching success, time to hatch, time for larvae to swim-up, and post-hatch growth and survival Exposure period

. . 88 day(s) Unit : ,FlC?Jl NOEC : >= 3.7

- measured/nominal LOEC : > 3.7

- measured/nominal Analytical monitoring : yes Method

. . OECD Guide-line draft “Early Life Stage Test (ELS-Test)’ Year : 2000 GLP : yes Test substance .

. as prescribed by 1.l

- 1.4
Attached document : Fish Early Life Stage In Rainbow Trout (Oncorhynchus mykiss) (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current OECD guidelines and Good Laboratory Practices.
Rainbow trout embryos were exposed to nominal HBCD water concentrations of 0.43, 0.85, 1.7,3.4 and 6.8 ug/L. The top two doses represent gamma HBCD’s water solubility (3.4 ugR) and two times gamma HBCD’s water solubility (6.8 ug/L). A negative control and solvent control group were also included. Mean measured concentrations (LC/MS with heated nebulizer operated inthe selective ion monitoring mode) were 0.25, 0.47, 0.83, 1.8 and 3.7 ug/L. This method was designed to monitor for all3 HBCD diastereomers; however, the trace residues of the alpha and beta diastereomers evident inthe water samples were below the established limits of quantitation. Comparison ofthe chromatograms from study initiation through study termination showed that the relative distribution of the HBCD diastereomers remained constant during the definitive study, and the gamma diastereomer measured results were consistent throughout the study. Hatching success, time to hatch, time for larvae to swim-up, and post-hatch growth and survival were evaluated during the 88-day test. Rainbow trout 45 I 88

## 4. Ecotoxicity Id Date 25637-99-4 31.03.2005

exposed to HBCD at mean measured concentrations up to 3.7 ug/L (nominal concentration = 6.8 ug/L or twice HBCD’s water solubility) for a 27-day hatching period and 61 days post-hatch showed no effects on hatching success, time to swim-up, larval survival, fry survival or growth. Consequently, HBCD was not chronically toxic to rainbow trout at concentrations at or above its limit of solubility. The NOEC for this study was 3.7 ug/L or 6.8 ug/L nominal (twice gamma HBCD’s water solubility). The low-effect-concentration (LOEC) and maximum acceptable toxicant concentration (MATC) could not be determined due to absence oftoxicity, but were considered >3.7 ug/L or >6.8 ug/L nominal (> twice gamm HBCD’s water solubility).
Drottar et al. 2001. Hexabromocyclododecane (HBCD): An early life-stage toxicity test with the rainbow trout (Onchorhynchus mykiss). Project No.: 439A-112. Wildlife international, Ltd. Easton, MD. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 29.12.2004 (10)

4.5.2 CHRONIC TOXlClTV TO AQUATIC INVERTEBRATES
Species : Daphnia magna (Crustacea) Endpoint

. . other: survival, reproduction and growth Exposure period : 21 day(s) Unit : Kd NOEC

. . = 3.4

- measured/nominal LOEC : = 5.6

- measured/nominal Analytical monitoring : yes Method : EPA OTS 797.1330 Year : 1998 GLP

. . Yes Test substance : as prescribed by 1.1

- 1.4
Attached document : Daphnia magna Life Cycle (21 Day) (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD and GLP guidelines.
Nominal test concentrations were 0.85, 1.7, 3.4, 6.8 and 13.6 ug HBCD/L water; dose levels were based on gamma HBCD’s water solubility, 3.4 ug/L. Measured test concentrations (LC/MS with negative ion atmospheric pressure ionization) were 0.87, 1.6, 3.1, 5.6 and 11 ug HBCD/L water (based on the gamma stereoisomer).
No statistically significant effects on survival, reproduction or growth of Daphnia magna were seen at HBCD concentrations <3.1 ug/L (measured). Thus, HBCD’s no effect concentration (NOEC), based on survival, reproduction and growth, to daphnia magna for 21 days was equivalent to HBCD’s water solubility. The measured NOEC inthis study was 3.1 ug/L and corresponded to a nominal HBCD concentration of 3.4 ug/L, e.g. gamma HBCD’s water solubility. The lowest observed effect concentration (LOEC) and the maximum acceptable toxicant concentration (MATC) based on survival, growth and reproduction were greater than HBCD’s water solubility. The LOEC, 5.6 us/L, corresponded to nominal concentrations twice gamma HBCD’s water solubility. The effect seen at this dose level was a reduction in length. Survival and reproduction at the 5.8 ug/L dose level were not affected. The MATC, 4.2 ug/L, was calculated 46188

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

as the mean of the NOEC and the LOEC. The MATC was greater than gamma HBCD’s water solubility.
Drottar K. and Krueger H. 1998. Hexabromocyclododecane (HBCD): Flowthrough life-cycle toxicity test with the cladocema (Daphnia magna). Project No.: 439A-108. Wildlife International, Ltd. Easton, MD. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 29.12.2004 (18)

4.6.1 TOXICITY TO SEDIMENT DWELLING ORGANtSMS

Species : Hyalella Endpoint : other: survival and growth, 5% TOC Exposure period : 28 other: days Unit : mg/kg sediment dw NOEC : = 1000

- measured/nominal LOEC : > 1000

- measured/nominal Method : other:ASTM E 1706-95b, OPPTS 850.1735 Year : 2003 GLP : yes Test substance : as prescribed by 1.1

- 1.4
Attached document : Prolonged Sediment Study with Hyalella azteca, 5% TOC (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. The definitive study was performed according to current ASTM, OECD and GLP guidelines.
Non-GLP exploratory range-finding studies were performed with three freshwater species associated with sediment: oligochaetes (Lumbriculus variegatus), chironomids (Chironomus riparius) and amphipods (Hyalella azteca). Allthree species were tested at 50, 100, 500, 1000 mg/Kg dry sediment intwo different types sediments, one with a2% organic carbon content and the other with a5% organic carbon content. Based on the results ofthe range-finding studies, the amphipods were found to be the most sensitive species in both sediment types, with clear effects inthe 500 mg/Kg treatment group. This reports the results of HBCD exposure inthe 5% organic content sediment in Hyalella azteca. Asimilar study in2% organic carbon was also performed. Groups of amphipods were exposed to a geometric series ofsix test concentrations and a negative control for 28 days under flow-through test conditions. Eight replicate test compartments were maintained ineach treatment and control group, with 10 amphipods in each test compartment, for atotal of 80 amphipods per test concentration. Each test compartment contained a quantity of sediment and overlying water. Additional replicates were added inthe control group, low and high treatment groups for analytical sampling of water and sediment. The ‘analytical’ replicates sampled on Day 0 contained no amphipods, while amphipods were added at test initiation to the “analytical” replicates sampled on Day 7 and at test termination.
Nominal test concentrations were 31, 63, 125,250, 500 and 1000 mg/Kg of sediment based on the dry weight of the sediment. The results ofthe study are based on the nominal test concentrations. Overlying water, pore water and sediment samples were collected and analyzed from the “analytical replicates” of the control group and the lowest and highest test 47188

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

concentrations. The collection and analysis were done approximately ten minutes after the addition of test organisms to the test system on Day 0, on Day 7 and at the end of the test. Results of the analyses were used to confirm the lowest and highest test concentrations. Analysis of HBCD concentrations inthe sediment, pore water and overlying water samples collected during test confirmed that the test article tended to remain insediment and not move into the pore water or overlying water. Concentrations insediment at the highest dose level ranged from 78.2

- 122% of nominal, while the lowest dose level ranged from below the limitof quantitation (12.5 mg/kg) to 197% of nominal. Alloverlying water samples contained no detectable HBCD. Pore water sample from the low concentration were also below the limit of quantitation, but those from the highest dose level, 1000 mg/kg sediment, had HBCD concentrations inthe low ppm range. These values were allwell above HBCD’s water solubility and were believed the result of small particles of HBCD being extracted out of the pore water, artificially inflating the reported values. Observations of amphipods in individual replicates appeared normal, with some mortality inthe 0,31,63, 125,250 and 500 mg/kg groups. Fungal growth was noted inall replicates.
The mean number of amphipods inthe negative control, 31, 63, 125, 250, 500 and 1000 mg/Kg treatment groups attest termination was 9.1, 8.6, 5.9, 6.1, 7.0, 8.5 and 9.1, respectively. The mean numbers of amphipods inthe 63, 125 and 250 mg/Kg treatment groups were found to be statistically different (p < 0.05) from the negative control group. Survival inthe 31, 500 and 1000 mg/Kg treatment groups was similar to the control group and any differences were not statistically significant (p>O.O5). Since the percent reduction inthe number of organisms present at test termination in comparison to the negative control group was less than 50% inall treatment groups, the 28-day EC50 value was estimated to be greater than 1000 mg/Kg of dry sediment, the highest concentration tested. The percent reduction from the control inthe 63, 125, and 250 mg/Kg treatment groups was 35.2, 33.0, and 23.1%, respectively. The mortality inthese groups was moderate and there were clearly no effects at the 500 and 1000 mg/Kg treatment levels. Therefore, the mortality observed inthe middle test concentrations was not considered treatment related since there was no evidence of a concentration dependent response. The average dry weight per amphipod inthe negative control group was 0.19 mg. The average dry weight per amphipod inthe 31, 63, 125, 250, 500 and 1000 mg/Kg treatment groups was 0.17, 0.26, 0.22, 0.20, 0.19 and 0.19 mg, respectively. The dry weights were not significantly different (p>O.O5) from the negative control weights, and any differences were not concentration-dependent. Therefore, there were noapparent effects on growth (dry weight) observed at test termination.
The 28-day EC50 value for amphipods (Hyalella azteca) exposed to hexabromocyclododecane insediment was >lOOO mg/Kg dry weight of sediment, the highest nominal concentration tested. Determination of the lowest-observed-effect-concentration (LOEC) and the no-observed-effectconcentration (NOEC) was based on an evaluation of the survival and growth (dry weight) data. The most sensitive parameter inthis study was survival. Based on the results of this study, the LOEC was >lOOO mg/Kg dry weight of sediment and the NOEC was 1000 mg/Kg dry weight of sediment.
Thomas J et al. 2003. Hexabromocyclododecane (HBCD): A Prolonged Sediment Toxicity Test with Hyalella azteca Using Spiked Sediment with 5% Total Organic Carbon. Project Number: 439A-120. Wildlife International, Ltd. Easton, MD. 48188

## 4. Ecotoxicity

Reliability Flag 2812.2004 Species Endpoint Exposure period Unit NOEC NOEC Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

: (1) valid without restriction Critical study for SIDS endpoint

. Hyalella

. other: survival and growth, 2% TOC 28 other: days : mg/kg sediment dw : = 1000

- measured/nominal : > 1000

- measured/nominal

.

. other: ASTM E 1706-95b, OPPTS 850. 1735

. 2003

. : 

## (19)

: ProlongedSediment Studywith Hyalellaazteca, 2%TOC (Sponsor:ACC Brominated FlameRetardant Industry Panel)
Thetest articlewasa compositeof3 lotsofcommercialHBCDproduced byAlbemarle Corporation, Great LakesChemicalCorporation, and Ameribrom, Inc. The definitive studywasperformed accordingto current EPA, OECD andGLP guidelines.
Non-GLP exploratory range-findingstudieswere performedwiththree freshwater speciesassociatedwith sediment:oligochaetes(Lumbriculus variegatus), chironomids(Chironomusriparius)andamphipods(Hyalella azteca). All three speciesweretested at50, 100, 500, 1000mg/Kgdry sedimentintwo different types sediments,onewith a2%organiccarbon content andthe otherwith a5% organiccarbon content. Basedonthe resultsofthe range-findingstudies,the amphipodswerefoundtb bethe mostsensitive speciesinbothsedimenttypes, with cleareffects inthe500 mg/Kgtreatment group.Thisreportsthe resultsof HBCDexposure inthe 5% organiccontent sedimentinHyalellaazteca. Asimilarstudy in5% organic carbonwasalsobeenperformed. Groups ofamphipodswere exposedto a geometricseriesofsixtest concentrations anda negative controlfor 28days underflow-throughtest conditions. Eight replicatetest compartmentswere maintainedineach treatment andcontrol group,with 10amphipodsineachtest compartment, for atotal of80amphipodspertest concentration. Eachtest compartment contained a quantity ofsedimentandoverlying water. Additional replicates were added inthe control group, lowand hightreatment groupsfor analytical samplingofwater andsediment.The “analytical” replicates sampledon Day0contained noamphipods,whileamphipodswere added attest initiationtothe “analytical” replicatessampledon Day7 andattest termination.
Nominaltest concentrations were 31,63, 125,250, 500and 1000mg/Kgof sedimentbasedonthe dryweight ofthe sediment.The resultsofthestudy are basedonthe nominaltest concentrations. Overlying water, porewater andsedimentsampleswerecollected andanalyzed fromthe “analytical replicates” ofthe control group andthe lowest andhighesttest concentrations. The collection andanalysiswere doneapproximatelyten minutesafter the additionoftest organismstothe testsystemon Day0,on Day7 and atthe endofthetest. Resultsofthe analyseswere usedto confirmthe lowestand highesttest concentrations. Analysis of HBCDconcentrations inthe sediment,porewaterandoverlying watersamplescollected duringtest confirmedthat thetest articletendedto remaininsedimentandnot move intothe porewateroroverlying water. Concentrations insedimentatthe highestdoselevel rangedfrom 82.8 - 115% of nominal,whilethe lowestdoselevel rangedfrom49.5to 125%of 49J88

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

nominal. All overlying water samples contained no detectable HBCD. Pore water sample from the low concentration were also below the limitof quantitation, but those from the highest dose level, 1000 mg/kg sediment, had HBCD concentrations inthe low ppm range. These values were all well above HBCD’s water solubility and were believed the result of small particles of HBCD being extracted out of the pore water, artificially inflating the reported values. Observations of amphipods in individual replicates appeared normal, with some mortality inthe 0,31 and 1000 mg/kg groups. Fungal growth was noted inall replicates.
The mean number of amphipods inthe 0,31,63, 125,250,500 and 1000 mg/kg groups at test termination was 7.4, 5.9, 7.8, 5.4, 6.9, 7.3, and 5.8, respectively. No statistical difference between treated and control groups were detected. The average dry weight per amphipod inthe 31, 63, 125, 250, 500 and 1000 mg/kg treatment groups was 0.10, 0.13, 0.14, 0.12, 0.17 and 0.14, respectively, and were not significantly different from control (P>O.O5).
Based on survival and growth (dry weight) data, the 28-day EC50 was > 1000 mg/kg of dry sediment, the highest concentration tested. The LOEC was > 1000 mg/kg and the NOEC was 1000 mg/kg dry weight of sediment.
Thomas S, Krueger H and Kendall T. 2003. Hexabromocyclododecane (HBCD): A Prolonged Sediment Toxicity Test with Hyalella azteca Using Spiked Sediment with 2% Total Organic Carbon. Project Number: 439A119B. Wildlife International, Ltd. Easton, MD. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 29.12.2004 (20)

4.6.2 TOXICITY TO TERRESTRIAL PLANTS

Species Endpoint Exposure Unit NOEC Method Year period : : : : : : : other terrestrial plant other: seedling emergence and growth 21 day(s) mg/kg soil dw >= 5000

- measured/nominal other: OPPTS 850.4100, 850.4225 2002 GLP Test substance : : yes as prescribed by 1.l

- 1.4
Attached document : Seediling (Sponsor: Emergence and Growth in Six Plant Species ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to EPA OPPTS 850.4100 and 850.4225; OECD Proposed for Revision of Guidleine 298; EPA/OECD Good Laboratory Practices.
The purpose of the study was to determine the effects of Hexabromocyclododecane (HBCD) on the seedling emergence and growth of six species of non-target plants. The experimental design for this study consisted of a negative control and five treatment groups. Each group had four replicate pots with ten seeds planted in each pot. Application of test concentrations of HBCD was made by soil incorporation to each treatment group prior to the planting of seeds. The nominal test substance 50188

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

concentrations were 0,40, 105, 276, 725, 1,904, and 5,000 mg of HBCD per kilogram of dry soil (mg HBCD/kg). The mean measured test levels were 0 (Negative Control), 31.3, 97.8, 297, 764, 2,230, and 6,200 mg HBCD/kg dry soil.
Seeds were impartially assigned to prelabeled growth pots on the day of test initiation. The replicate pots were placed ina randomized block design on a greenhouse table after planting. Observations of emergence were made on Days 7, 14, and 21. A general assessment of seedling condition was made on Day 7, while observations of height, shoot dry weight, and assignment of plant condition scores were made only on Day 21.
There were no apparent effects on any endpoint for any of the six species tested. Statistical analyses indicated that there were no significant differences (Dunnett’s test, p>O.O5) between the control and treatment group mean emergence, survival, height, or weight for corn, cucumber, ryegrass, soybean and tomato. On day 21, onion showed astatistically significant difference (Dunnett’s test, ~~0.05) between the control and the 276 mg HBCD/kg treatment group mean survival. This significant difference was not considered dose-responsive, and not attributed to treatment, as no statistical differences were noted at higher concentrations tested (Dunnett’s test, ~~0.05). There were no statistically significant differences (Dunnett’s test, p>O.O5) between the control and signs of treatment-related phytotoxicity observed on seedlings of any species at any test concentration.
No effects from soil incorporation of HBCD were observed on seedling emergence, survival, or growth for any of the six plant species tested. Therefore, the NOEC for emergence and growth of allseedlings inthis study was determined to be 5,000 mg of HBCD/kg, which was the highest nominal soil concentration tested.
Porch etal. 2002. Hexabromccyclododecane (HBCD): A Toxicity Test to Determine the Effects of the Test Substance on Seedling Emergence of Six Species of Plants. Project Number: 439-l 03. Wildlife International, Ltd. Easton, MD. Reliability : (1) valid without restriction Flag : Risk Assessment, Critical study for SIDS endpoint 28.12.2004 (21)

4.673 TOXICITY TO SOIL DWELLING ORGANISMS



## Type : artificial soil Species : Eisenia fetida (Worm (Annelida), soil dwelling) Endpoint

. . mortality Exposure period : 28 day(s) Unit . mg/kg soil dw NOEC : >= 4190

- measured/nominal LC50 : > 4190

- measured/nominal Method : EPA OPPTS 850.62 Year . 2002 GLP : Yes Test substance : as prescribed by 1.l

- 1.4 Attached document : Earthworm (Eisenia fetida) Survival and Reproduction (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, 51188

## 4. Ecotoxicity

Reliability Flag 28.12.2004 Type Species Endpoint Exposure period Unit NOEC Id 25637-99-4 Date 31.03.2005

OECD and GLP guidelines.
The artificial soil used inthis study was characterized as sandy loam with an 80% sand, 8% silt, and 12% clay content. Nominal test concentrations were 0, 78.5, 157, 313, 625, 1,250, 2,500 and 5,000 mg HBCD/kg of dry soil. Mean measured concentrations at day 28 were cl .28 (Control), 61.2, 145, 244, 578, 1,150, 2,180, and 4,190 mg HBCD/kg ofdry soil. Mean measured concentrations at day 56 were 56 Days: cl.35 (Control), 51.5, 128,235, 543, 1,070, 2,020, and 3,990 mg HBCD/kg ofdry soil. Measured tissue concentrations at day 28 were ~0.200 (control), 3.40, 7.32, 16.8, 15.3, 53.0, 71.2, and 150 mg HBCD per gram oftissue.
After 28 days of exposure to HBCD, percent mortality ofthe adult worms was 0, 5, 0, 0, 0, 0, 3, and 0% inthe 0, 61.2, 145, 244, 578, 1,150, 2,180, and 4,190 mg HBCD/kg groups, respectively. Allof the live earthworms were normal inappearance and behavior. No abnormal burrowing or avoidance behaviors were recorded during the first 60 minutes of testing.
The control worms gained an average of 0.418 g per replicate or 10% in replicate mass during the 28 days of adult worm exposure. The mean replicate weight gain of the surviving treatment animals ranged from 0.018 to 0.808 grams. These gains represented average increases of 0.4 to 19% in replicate animal mass over the initial 28 days of exposure. The average reproduction inthe control replicates was 72 juveniles per replicate. The coefficient of variation for the control data was 16%. The average reproduction was 61, 60,49, 31, 26,26, and 30 juveniles per replicate for treatment levels 51.5, 128, 235, 543, 1,070, 2,020, and 3,990 mg HBCD/kg, respectively. A statistically significant (p f 0.05) reduction in reproductive output occurred at treatment levels 3235 mg HBCD/kg. Using the estimated soil Koc and the formula provided inthe European Union’s Technical Guidance Document, HBCD’s calculated BCF in earthworms is:

BCFearthworm = Cearthworm/Csoil

= (Kearthworm-porewater)(RHOsoil)(l0+3)/Ksoil-water (0.15 mg/kg)(l.25 x 10+5)/4,190 mg/kg dry soil =4.5. Thus, HBCD did not bioconcentrate inthe earthworm.
The 28-Day EC50 (survival) was >4,190 mg/kg. The 28-Day NOEC (survival) was 4,190 mglkg. The 56-Day EC50 (reproduction) was 771 mglkg with 95% confidence limits of 225 to 4,900 mglkg. The 56-Day NOEC (reproduction) was 128 mg/kg.
Aufterheide et al. 2002. Effect of Hexabromocyclododecane on the Survival and Reproduction ofthe Earthworm, Eisenia fetida. ABC Study No. 47222. ABC Laboratories, Inc., Columbia, Missouri; Wildlife International, Inc., Easton, MD.

: (1) valid without restriction : Risk Assessment, Critical study for SIDS endpoint (11)

: artificial soil : Eisenia fetida (Worm (Annelida), soil dwelling) : other: reproduction : 56 day(s) : mg/kg soil dw : = 128

- measured/nominal 52188

## 4. Ecotoxicity Id 25637-99-4 Date 31.03.2005

EC50 Method Year GLP Test substance
Attached document .

. : : = 771

- calculated EPA OPPTS 850.62 2002 : : yes as prescribed by 1.l

- 1.4

: Earthworm Brominated (Eisenia fetida) Survival and Reproduction Flame Retardant Industry Panel) (Sponsor: ACC

The test article was a composite by Albemarle Corporation, Great Ameribrom, Inc. This study was OECD and GLP guidelines. of 3 lots of commercial HBCD produced Lakes Chemical Corporation, and performed according to current EPA,

The artificial soil used inthis study was characterized as sandy loam with an 80% sand, 8% silt, and 12% clay content. Nominal test concentrations were 0, 78.5, 157, 313, 625, 1,250, 2,500 and 5,000 mg HBCD/kg of dry soil. Mean measured concentrations at day 28 were e1.28 (Control), 61.2, 145,244, 578, 1,150,2,180, and 4,190 mg HBCD/kg of dry soil. Mean measured concentrations at day 56 were 56 Days: cl .35 (Control), 51.5, 128,235, 543, 1,070,2,020, and 3,990 mg HBCD/kg of dry soil. Measured tissue concentrations at day 28 were co.200 (control), 3.40, 7.32, 16.8, 15.3, 53.0, 71.2, and 150 mg HBCD per gram of tissue.
After 28 days of exposure to HBCD, percent mortality ofthe adult worms was 0, 5,0, 0, 0, 0, 3, and 0% inthe 0, 61.2, 145,244, 578, 1,150, 2,180, and 4,190 mg HBCD/kg groups, respectively. Allof the live earthworms were normal inappearance and behavior. No abnormal burrowing or avoidance behaviors were recorded during the first 60 minutes of testing.
The control worms gained an average of 0.418 g per replicate or 10% in replicate mass during the 28 days of adult worm exposure. The mean replicate weight gain of the surviving treatment animals ranged from 0.018 to 0.808 grams. These gains represented average increases of 0.4 to 19% in replicate animal mass over the initial 28 days of exposure. The average reproduction inthe control replicates was 72 juveniles per replicate. The coefficient of variation for the control data was 16%. The average reproduction was 61, 60,49, 31, 26, 26, and 30juveniles per replicate for treatment levels 51.5, 128, 235, 543, 1,070, 2,020, and 3,990 mg HBCD/kg, respectively. A statistically significant (p f 0.05) reduction in reproductive output occurred at treatment levels 3235 mg HBCD/kg. Using the estimated soil Koc inTable 1and the formula provided inthe European Union’s Technical Guidance Document, HBCD’s calculated BCF in earthworms is:
BCFearthworm = CearthwormICsoil porewater)(RHOsoil)(10+3)/Ksoil-water = (Kearthworm-

(0.15 mg/kg)( 1.25 x 10+5)/4,190 mg/kg dry soil =4.5. Thus, HBCD did not bioconcentrate inthe earthworm.
The 28-Day EC50 (survival) was >4,190 mg/kg. The 28-Day NOEC (survival) was 4,190 mg/kg. The 56-Day EC50 (reproduction) was 771 mg/kg with 95% confidence limits of 225 to 4,900 mg/kg. The 58-Day NOEC (reproduction) was 128 mg/kg. Aufterheide et al. 2002. Effect of Hexabromocyclododecane on the Survival and Reproduction of the Earthworm, Eisenia fetida. ABC Study No. 47222. ABC Laboratories, Inc., Columbia, Missouri: Wildlife 53188

## 4. Ecotoxicity Id Date 25637-99-4 31.03.2005

Reliability Flag 29.12.2004

4.6.4 TOX. TO OTHER international, Inc., Easton, MD.

: (1) valid without restriction : Risk Assessment, Critical study

NON MAMM. TERR. SPECIES for SIDS endpoint (22)

4.7 BIOLOGICAL EFFECTS MONlTORlNG

4.0 BlOTRANSlfORMATlON AND KlNETlCS

4.9 ADDIRONAL REMARKS

54188

## 5. Toxicity id 25637-99-4 Date 31.03.2005

5.0 TOXICOKINETICS, METABOLISM AND DiSTFiiBUTlON

In Vitro/in vivo : invivo Type Species : Toxicokinetics : rat Number of animals Males Females Doses Males . . Females .

. Vehicle .

. Method : other Year : GLP : no data Test substance : other TS
Attached document : From “Toxicological Risks of Selected Flame Retardants”, National Academy Press, Washington, DC, 2000, pages 54-55:
“Toxicokinetics
No human data on the toxicokinetics of HBCD were located for any route. No toxicokinetic studies via the dermal or inhalaiton exposure routes were reported in experimental animals. However, ina report by Dean and Leong (1977), rats exposed dermally to a high dose of HBCD insaline experienced diarrhea and slight weight loss. This finding indicates that at least some absorption occurs via the dermai route.
Inan unpublished study by Vesicol Chemical Corporation (1980), rats administered asingle oral dose of 1.93 mg of radiolabeled HBCD elimenated 86% of the dose within 72 hr. (The total dose administered was 7-9 mg/kg body weight.) Absorption from the gastrointetinal tract reportedly occurred rapidly, with a half-life of 2 hr. However, the amount of the absorbed fraction was not reported. HBCD was repotted to be rapidly metabolized and eliminated inthe feces and urine following absorption, with 70% of the administed radioactivity eliminatd inthe feces and another 16% eliminated inthe urine 72 hr after dosing. A two-compartment model was constructed, with non-adipose tissues inone compartment and adipose tissue inthe other. Elimination from the adipose compartment was reported to be slower than elimination from the non-adipose compartment, althougth elimination half-times were not provided inthe review. in another stuy by Arita et al. (Marcia Hardy, Albemarle Corporation, Pers. Commun., August 3, 1999), HBCD was orally administered to male Wistar rats (number not reported) inolive oilat500 mg/kg-d for 5 d. HBCD was found to be present only inadipose tissue, and in none of the other organs examined (i.e., spleen, pancrease, liver, kidneys, and heart). HBCD was found to be excreted inthe feces, with an average of 32-35% of the cumulative administered dose excreted. No HBCD was found inthe urine. Although differences instudy design, including the test vehicle and the analytic methods used, may account for some of the difference inthe results, both studies by Vesicol Chemical Corporaiton (1980) and Arieta et al. (Marcia Hardy, Albemarle Corporation, Pers. Commun., August 3, 1999) suggeest that following acute oral doses, HBCD is rapidly absorbed from the gastrointestinal tract, distributed primarily to body fat, and eliminated rapidly, primarily inthe feces.”
For additional details on these studies, please see the following records. 27.12.2004 (23) 

# I 55188 I

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

In Vitro/in vivo : Invivo Type : Toxicokinetics Species : rat Number of animals Males : 4 Females : 0 Doses Males : 500 mg/kg bw/d Females : Vehicle : other: olive oil Route of administration : gavage Exposure time : 5day(s) Product type guidance : Decision on results on acute tox. tests : Adverse effects on prolonged exposure : Half-lives : l@: 2? 3? Toxic behaviour : Deg. product : Method : other Year : 1963 GLP : no data Test substance : other TS
Attached document : The excretion of HBCD in urine and feces as well as its distribution to various organs was investigated. The test article was “Pyroguard SR-103”, manufactured by Daiichi Kogyo Seiyaku K.K.
A fine suspension of SR-103 inolive oilwas prepared by miing well to homogeneity ina mortar. The dose was 500 mglkgld for 5 consecutive days (n= 4 male Wistar rats). The dose volume was 100 mg/ml. Urine and feces were coleected separately by housing the rats in glass cages. Twenty-four hours after the last administration, the rats were sacrifices and the spleen, liver, pancreas, kidneyss, heart and fatty tissue collected. Gas chromatography with an FID detector was used for quantification. The limit of quantification was about 5 ug/ml in urine and about 20 ug/ml inthe fecal and organ homogenates.
The average daily rate of excretion inthe feces was 29-37% of the dose. The cummulative excretion was roughly constant at 32-35% with respect to the cummulative administered amount. Urinary excretion was not observed. No evidence for the presence of metabolites was observed in urine or feces.
A separate study with isolated intestinal loop (upper jejunum) indicated about 12% of the dose was detectable inthe intestinal tissue and the amount remaining inthe lumen loop “small”.
The test article was detected only inthe adipose tissue after dosing for 5 days. The level inadipose tissue was 0.3-0.7 mg/g fat.
R. Arita, K. Miyazaki and S. Mure. 1983. Metabolic test of hexabromocyclododecane. Department of Pharmacy, Hokkaido University Hospital. Japan. 28.12.2004 (24) In Vitro/in vivo : In vivo Type : Toxicokinetics Species

. . rat 56188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

Number of animals Males : Females : Doses Males : 7-9 mg/kg Females : 7-9 mg/kg Vehicle : other: olive oil Route of administration : gavage Exposure time : Product type guidance : Decision on results on acute tox. tests : Adverse effects on prolonged exposure : Half-lives : l? 27 hour(s) 2? 3’(‘: Toxic behavlour : Deg. product

. . Method : other Year : 1980 GLP : no data Test substance : other TS: 14C; probably gamma Attached document : Asingle oral dose (7-9 mg/kg) of 14C-HBCD was administered to male (n=2) and female (n=8) rats. Based on the starting material and the melting point of the final product described inthe report, the test article appeared to be composed of the gamma isomer. The rats were sacrificed 8,24,48 and 72 hours (females) and 48 hours (males) after dosing. One female rat served as control. Urine and feces were collected daily, blood samples from 4animlas were dollected during the first 24 ours. Tissue samples were collected at the time of sacrifice.
Total 14C-activity was determined via liquid scintillation and the parent molecule and metabolites were distinguished by thin layer chromatography in extracts of feces and urine. HBCD appeared to be well absorbed from the gastrointestinal tract and extensively metabolized prior to elimination infeces (primary route) and urine. The estimated absorption half-life was 2 hours; peak radioactivity was detected in blood 4 hours post-dosing. The pharmacokinetics of the gamma stereoisomer appeared to follow an open twocompartment model. The central compartment was described as liver, lung, kidney, heart, muscle, gonads, uterus, spleen, and brain; the peripheral compartment was described as adipose tissue. Roughly 80

- >90% of the gamma stereoisomer was eliminated within 3 days following a single oral dose, with an apparent half-life of elimination of 27 hours. Only metabolites were detected infeces and urine

- no parent molecule was detected. Thus, the gamma isomer appears to be extensively metabolized prior to excretion. In contrast, only unmetabolized gamma isomer was detected inadipose tissue.
In females at 8, 24, 48 and 72 hours post-dosing, the total 14Cactivity detected intissues of female rats was -43, 24,18 and 17% of the dose, respectively. In male rats at 48 hours post-dosing (the only time point investigated in males), the 14Cactivity was -10% ofthe dose. Similarly, that detected infeces from female rats was -4, 65, 54 and 77% at 8,24, 48 and 72 h. Urine contained 0.1, 6, 18and 15% of the dose at the same time intervals. Feces from male rats at 48 hours contained -94% of the dose while urine from male rats contained -15%. At 48 hours post-dosing approximately 81% of the dose was detected infeces and urine of female rats. Thus, at 48 hours post-dosing approximately 86% of the dose was recovered inthe 

### tissues, feces and urine from female rats whereas 119% was recovered from males. The lower 48 hour recovery from this group of 57188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

female rats is largely accounted for by the lower fecal content (43%) of radioactivity collected during the O-24 hours post-dosing. By 48 hours post-dosing, females sacrificed at24 hours, had atotal fecal 14Ccontent of 65% ofthe dose, and females sacrificed at 72 hours had atotal fecal 14Ccontent of 62% of the dose. Substituting the average value, 63%, from these two groups for the 43% value used to calculate overall recovery, the total per cent of the dose accounted for at 48 hours becomes feces (74%) urine (17%) and tissues (18%) or 109% of the dose. Itappears likely that some unknown factor resulted inthe lower O-24 hour percent recovery from feces, and that percent of dose present intissues, feces and urine 48 hours post-dosing is similar in male and female rats. Caution is urged in interpreting this data due to the small sample size and the brief nature ofthe final report. C.C. Yu and Y. H. Atallah. Velsicol Chemical Corporation, Laboratory Report. September 17, 1980. 28.12.2004 (25)

5.1-l ACUTE ORAL TOXlCtTY

Type : LD50 Value . >lOOOO- mglkgbw Species : rat Strain : other: albino Sex : male/female Number of animals

. . 10 Vehicle : other: corn oil Doses

. . 0, 10000 Method

. . other: Hagan etal. 1959 Year : 1978 GLP : no Test substance : no data Reliability : (2) valid with restrictions Flag : Critical study for SIDS endpoint 28.12.2004 (26)

51.2 ACUTE INHALATION TOXICITY



### Type : LC50 Value : >200- mg/l Species

. . rat Strain : Wistar Sex : male/female Number of animals : 10 Vehicle

. . other: material was used as received Doses . 200 mg/l Exposure time : 1 hour(s) Method : other Year : 1978 GLP : no Test substance : no data
Attached document : Inhalation LC50
Albino rats in groups of 10 (5M:5F), 233-292 g, were exposed to a concentration of200 mg/L, the highest possible chamber concentration, for
58188

## 5. Toxicity Id Date 25637-99-4 31.03.2005

1 hour and observed for two changes were observed. weeks. No deaths occured, and no gross

Reliability Flag : : The material (2) valid with Critical study was not toxic by inhalation restrictions for SIDS endpoint under the conditions of this test.

26.12.2004

5.1.3 ACUTE DERMAL TOXfCfTY (26)

as received mg/kg bw 

### Type Value Species Strain Sex Number of animals Vehicle Doses Method Year GLP Test substance
Attached document

Reliability Flag 28.12.2004

5.1.4 ACUTE TOXlClTY,

5.2.1 SKIN IRRlTATlON

Species Concentration Exposure Exposure time Number of animals Vehicle PDII Result Classification Method Year GLP Test substance : LD50 : > 8000

- mg/kg bw : rabbit : New Zealand white : male/female : 6 : other: material used

.

. 500,2000,5000,8000 : other: Hagan 1959 : 1978 : no : no data

: Acute Dermal Toxicity in Rabbits Following a 1Cday range finding, albino rabbits in groups of 6 (3M:3F), abraded, 1.88-2.07 kg bw, highest dose mechanically possible, single application dermally under occulded patch, observed for fourteen days. Material used as received. No animals died on test. l/2

: : The test material was not toxic dermally this test. (2) valid with restrictions Critical study for SIDS endpoint to rabbits under the conditions of

(26)
OTHER ROUTES

: rabbit

: -5 9

. Occlusive

.

: 24 hour(s)

.

. 6

. other: material

.

: 0

.

. not irritating

: not irritating

: other: Draize

: 1978

: no

I

. no data used as received

et al. 1944

Attached document : Primary Dermal Irritation in Rabbits
A group of albino New Zealand rabbits were used. Teh test method was 59188

## 5. Toxicity

Reliability Flag 28.12.2004 Species Concentration Exposure Exposure time Number of animals Vehicle PDII Result Classification Method Year GLP Test substance
Attached document

Flag 1512.2004

5.2.2 EYE IRRITATION

Species Concentration Dose Exposure time Comment Id 25637-99-4 Date 31.03.2005

essentially that of Draize et al. 1944. Briefly, 0.5 g of test material was applied to clipped aras of intact and abraded skin. The abrasions were longitudinal epidermal incisions sufficiently deep to penetrate the stratum corneum, but not so deep as to destroy the integrity of the derma. Applications were made under occulusive patches (2” x 2” gauze, covered by adhesive tape). The entire trunk of each animal was then covered with an impermeable occlusive wrapping, and everted Elizabethan collars were placed on each animal. The wrapping and test material were removed 24 hours after application. The sites were individually examined and scored separately for erythema and dema at 24 and f72 hours. The mean scores for 24 and 72 hour gradings were averaged to determine final irriation indices.
The test material was not irritating. The Primary Irritation Index was 0.0. : (2) valid with restrictions : Critical study for SIDS endpoint

: rabbit : .5g : Occlusive : 4 hour(s) : 6

. - other: material : 0 : not irritating : not irritating : other: Draize : 1978 : no : no data (26)

used as received

et al. 1944

: Dermal Corrosion in Rabbits 49 CFR 173,240(a)(1)
A group of 6 albino New Zealand rabbits, 1,8-2.4 kg, were used. The test method was essentially that of Draize et al. (1944). Briefly, 0.5 g of the test material was applied to clipped areas of intact skin. Applications were made under occulusive patches (2’ x 2’ gauze, covered by adhesive tape). The entire trunk of each animal was then coverved with an impermeable occulusive wrapping, and the animal immobilized. The wrapping and test material were removed 4 hours following application. The sites were individually examined and scored for erythema and edema at 4 and 48 hours. The mean scores for each time period were averaged to determine the final irritation indices. Corrosiveness seen at4 and/or 48 hours alone indicates a corrosive material.
The test material was not corrosive. The Primary Irritation Index was 0.0. : Critical study for SIDS endpoint (26)

: rabbit : -1 9 : .l other: g : 24 hour(s) : rinsed after (see exposure time) 60188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

Number of animals Vehicle Result Classification Method Year GLP Test substance
Attached document

Flag 15.12.2004

5.3 SENSlTlZATlON



### Type Species Number of animals Vehicle Result Classification Method Year GLP Test substance
Attached document : 6 : other: material used as received : not irritating : not irritating : other: Draize et al. 1944 : 1978

.

. no

- no data

.
: Primary Eye Irritation in Rabbits (FHSA)
New Zealand rabbits without ocular defects were used. The method was essentially that of Draize et al. 1944. Briefly, the test substasnce was instilled inthe right eye of each animal; the left eye was untreated and served as a control. The treated eyes of all rabbits remained unwashed for 24 hours. Readings were made at 1,2, and 3 days after treatment. Additional reading were made on days 4-7 as needed.
Mild transient occular irritation was noted at day 1,which decreased progressively over days 2 and 3, and had completed dissapeared by day 4. The scores indicate this material is not irritating according to EU regulations. : Critical study for SIDS endpoint (26)

: Mouse local lymphnode assay : mouse : 6 : other: DMF : not sensitizing : not sensitizing : other: OPPTS 870.2600 : 2003 : yes : as prescribed by 1.1

- 1.4

: Contact sensistization potential via the local lymph node assay (including primary irritancy screen) using CBA/J mice. (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of equal parts of the commercial materials produced by Albemarle Corporation, Dead Sea Bromine Compounds, and Great Lakes Chemical Corporation. Its composition was 8.68% alpha, 6.12% beta and 85.10% gamma. This study was performed according to Good Laboratory Practices, OPPTS 870.2660, and OECD 429.
The Local Lymph Node Assay asseses the potential of test materials to cause contact sensitization by measuring hte lymphocyte proliferative responses from auricular lymh nodes following topical application of the test materials to mouse ears. Test materials that elicit a Stimulation Index (SI) >=3 (i.e., 3-fold greater proliferation than control animals) should be considered positive for dermal sensitization potential.
All mice received one of three concetntrations of HBCD (2%, 20% or 50% w/v) or DMF (dimethylformamide) on days l-3 (n=6 mice/group). HCB (alpha-hexyl cinnamaldehyde), a moderate contact sensitizer, was 61 I88

## 5. Toxicity

Reliability Flag 15.12.2004 Type Species Concentration

Number of animals Vehicle Result Classification Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

evaluated concurrently as a positve dermal sensitizstion control. The test materials were administered to the dorsal surface of both ears (25 ul/ear). On day 6, all mice received ano intravenous tail vein injection of phosphate buffered saline containing 20 uCi of 3H-thymidine. Uptake into the auricular lymph nodes draining the site of chemical application was measured 5 hours later. Body weight data were unremarkable and minor increases inear thichness were noted suggesting slight irritation following applications of 20% and 50% HBCD. There were no indications that HBCD possesses dermal sensitization potential. SI values were consistently around 1.Oat alldoses tested. Lymphocyte proliferation by DMF, vehihcle treated mice (2015 dpm) was higher than historical laboratory values commonly observed using acetone and olive oilas avehicle. This is not inconsistent with that reported inthe literature for this vehicle. HCA adminstrations (30% v/v) elicited prolifeation that waes 3-fold greater than that of vehicle controls thus dtecting the moderate contact sensitization potential inthis study. On the basis of these results, HBCD would not be considered to have contact sensitization potential.
Woolhiser Mand Anderson P. 2003. Hexabromocyclododecane: Contact sensistization potential via the local lymph node assay (including primary irriiancy screen) using CBA/J mice. Study ID 031013. Toxicology & Envionmental Research and Consulting. The Dow Chemical Company. Midland, Ml (1) valid without restriction Risk Assessment, Critical study for SIDS endpoint (27) Guinea pig maximization test gjinea pig 1 : Induction 5 % intracutaneous 2? Induction 250 mg occlusive epicutaneous 3? Challenge 250 mg occlusive epicutaneous
other: corn oil not sensitizing not sensitizing other: EPA 40CFR798.4100, OECD 408 1996 yes as prescribed by 1.1

- 1.4 HBCD Maximization Test in Guinea Pigs (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of equal parts of the commercial products manufactured by Albemarle Corporation, Dead Sea Bromine Group, and Great Lakes Chemical Corporation. Its composition was 8.5% alpha isomer, 6% beta isomer, and 79.1% gamma isomer. HBCD was administered in2stages (induction and challenge doses) to male guinea pigs (Group 1, n=lO; Group 2 n=20). The induction doses were administered to the interscapular region as intradermal injections (phase I) and topical applications (phase II). The challenge dose was administered as topical applications to the flanks of each animal. Phase I of the induction doses consisted of three pairs of intradermal injections of 1) Freund’s adjuvant and corn oil (50:50), 2) the test article at a concentration of 5% in corn oil,or 3) the test article at a concentration of 5% inthe 50:50 corn oil/Freund’s adjuvant solution which were adminsitered to each treated animal (Group 2). The control animals (Group 1) received the same regimen but the test article was omitted. After a period of 7 days, the treated animals (Group 2) received topical applications of neat test article (moistened in corn oil) on the previously 62188

## 5. Toxicity

Reliability Flag 15.12.2004 Type Species Number of animals Vehicle Result Classification Method Year GLP Test substance
Attached document

Reliability Flag 20.12.2004
Number of animals Vehicle Result Classification Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

treated interscapular sites. Following atwo week rest period, the challenge doses were administered. Each animal received a topical application of neat tet article (moistened with corn oil) on the left flank and an empty Hilltop Chamber on the right flank. Subsequent examination of the test sites indicated the test article produced no erythema or edema inany animal at 24,48,72,96 or 120 hours after the challenge dose. Based on the percentage of animals responding (0%) HBCD was consided a non-sensitizer.
M. Wenk. Hexabromocyclododecane Maximization Test in Guinea Pigs. 1996. M96A061.1X64. Microbiological Associates, Inc. Rockville, MD. (1) valid without restriction Risk Assessment, Critical study for SIDS endpoint (28)

. * Patch-Test

.

. human :

.

. : not sensitizing : not sensitizing

.

. other : 1972 : no data

.

. other TS

: The test samples were Tyvek T-12 with 10% HBCD. One inch squares of the test samples were applied to the arms of 10 men and to the arms or legs of ten women and held in place with Dermicel tape for six days. After atwo-week rest period, new patches were applied for 48 hours as a challenge test for skin sensitization. Skin under the patches was examined at two and six days after the first application and on removal ofthe challenge patch. No skin reactions were observed on any subject at any examination McDonnell, M. 1972. Haskell Laboratory Report No. 185-72. Haskell Laboratory for Toxicology and Industrial Medicine. (1) valid without restriction Critical study for SIDS endpoint (29)

. .

. : : :

. . : no data

.

. other TS

.

. Five sensitization studies have been conducted on HBCD; three inguinea pigs, one in mice, and one in human volunteers. The 1997 guinea pig maximization test performed by BFRIP was negative. Two studies inthe literature (Momma et al. Pharmacometrics, 1985, 29:981-986 and Nakamura et at. Contact Dermatitis, 1994,31:72-85) reported positive results; the test article appears to have been an HBCD product produced in Japan. The patch test in human volunteers was negative. The 2003 mouse local lymph node assay, performed by BFRIP to clarify these results, was negative.
63188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

The reason for the discrepancy between these results is not apparent. However, the negative results inthe 1997 BFRIP test using the highest possible concentration for topical induction and challenge, raise questions about the potential for HBCD to produce even a mild sensitization reaction in humans. The methodologies used inthese 3sensitization tests are provided inthe table below. Comparison of the methodology used in3 guinea pig skin sensitization studies conducted on HBCD. BFRIP Monma Nakamura 1995 1985 1994 INDUCTION

- ID

VOLUME 0.1 ml 0.05 ml Assume 0.05 ml ? CONCENTRATION 5% 0.05, 0.5, 5% 0.5, 5% DOSE 0.005 mg 0.000025, 0.00025, 0.0025 mg0.00025,0.0025 mg VEHICLE Corn oil Olive oil Olive oil INDUCTION -TOPICAL AMOUNT 500 mg 200 mg Assume 200 mg ? CONCENTRATION 100% 25% 25% DOSE 250 mg 50 mg 50 mg VEHICLE Corn oil*Vaseline Petrolatum CHALLENGE VOLUME/AMOUNT 500 mg 0.02 ml 0.1 ml CONCENTRATION 100% 0.005,0.05,5% 0.05,0.5,5% DOSE 250 mg 0.000001, 0.00001, 0.0001, 0.001 mg 0.00005, 0.0005, 0.005 mg VEHICLE Corn oil*Acetone Acetone

l Only moistened with corn oil.

McDonnell, M. 1972. Haskell Laboratory Report No. 185-72. Haskell Laboratory for Toxicology and Industrial Medicine) Momma et al. Pharmacometrics, 1985, 29:981-986. Nakamura et al. Contact Dermatitis, 1994, 31:72-85. M. Wenk. Hexabromocyclododecane Maximization Test in Guinea Pigs. 1996. M96A061.1 X64. Microbiological Associates, inc. Rockville, MD. Woolhiser M and Anderson P. 2003. Hexabromocyclododecane: Contact sensistization potential via the local lymph node assay (including primary irritancy screen) using CBA/J mice. Study ID 031013. Toxicology & Envionmental Research and Consulting. The Dow Chemical Company. Midland, Ml
Reliability 29.12.2004

5.4 REPEATED DOSE : (4) not assignable

TOXICITY (30) (31)

64188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005


### Type Species Sex Strain Route of admin. Exposure period Frequency of treatm. Post exposure period Doses Control group NOAEL LOAEL Method
Year GLP Test substance
Attached document : Sub-chronic : rat : male/female : Sprague-Dawley : gavage : 28 Days : Once daily : 14Days : 0, 125,350, 1000 mg/kg/d : yes, concurrent vehicle

. . = 1000

- mg/kg bw : > 1000

- ml/kg bw : OECD Guide-line 407 “Repeated Dose Oral Toxicity

- Rodent: 28day or 14-d Study” : 1996 : yes : as prescribed by 1.1

- 1.4

: A 26-day repeated dose oral toxicity study of HBCD in rats. (Sponsor: ACC Brominated Flame Retardant Industry Panel)
This study was conducted according to OECD and GLP guidelines. The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. HBCD, inthe vehicle corn oil,was administered orally by gavage to three groups of Sprague-Dawley Crl: CD BR rats for a period of 28 consecutive days. Dose levels were 125 (low), 350 (mid), or 1,000 (high) mglkglday, administered at dosage volume of 5 ml/kg. The test groups consisted of 6 males and 6females inthe 125 and 350 mg/kg/day groups and 12 males and 12females inthe 1000 mg/kg/day group. A concurrent control group comprised of 12 males and 12females received the vehicle, corn oil,for 28 consecutive days at a dosage volume of 5 ml/kg. At the end ofthe dosing period, 6 animals/sex/group were sacrificed and necropsied. The remaining 6animals/sex inthe control and 1000 mg/kg/day groups remained on-test untreated for a 1Cday recovery period. At the end of the recovery period, allanimals were sacrificed and necropsied.
Animals were observed twice daily for mortality and moribundity. Clinical signs were recorded daily. Body weight and food consumption were measured weekly. Functional observational battery and motor activity evaluations were performed during weeks -1 (pretest), 3, and 5 (recovery). Samples for hematology and serum chemistry evaluations were collected atthe primary (28 day) and recovery (42 day) sacrifices. Complete necropsies were performed on all rats. The brain, liver, kidney, heart, spleen, testes and epididymus or ovaries, adrenal glands, and thymus from allanimals were weighed at each sacrifice. Approximately 40 tissues were collected and preserved at each necropsy from allanimals. The following tissues were examined microscopically from the control and high dose animals: liver, kidney, heart, spleen, testes (males), prostate (males), seminal vesicles (males), epididymus (males), ovaries (females), adrenal glands, thymus, bone with marrow (sternebra), brain, stomach, ceoum, duodenum, ileum, jejunum, lymph node, peripheral nerve (sciatic), spinal cord, lung, trachea, uterus (females), urinary bladder, and all gross lesions. The lungs, liver, kidneys, stomach, thyroid, gross lesions and target organs were examined inalldose levels. Survival was not affected by administration of the test article. Allanimals survived to the scheduled sacrifice. Clinical signs observed during the study were nonspecific, low in incidence, non-dose-related and not considered related to test article. 65 I88

## 5. Toxicity Id 25637-99-4 

### Date 31.03.2005

Bodyweights,weight gainandfood consumptionoftreated animalswere comparedstatistically bysex andtreatment day totheir respective control groups (p< 0.05 or0.01) andwere notaffected bytreatment. No statistically significantdifferencesin bodyweight betweencontrol and treated animalswere detected withthe exception ofan increaseinmean female bodyweight inthe 350mg/kg/day group duringweek2of treatment. Meanfemalebodyweight atthattime pointwas 196gversus 179inthe control group. Nostatisticallysignificant differencesin body weight gainbetween control andtreated animalswere detectedwiththe exception ofa decreaseinmeanmalebodyweight gaininthe 1,000 mg/kg/day recovery groupduringweek 1of recovery. Mean malebody weight gainatthattime pointwas21 gversus 31 inthecontrol group; meanmalebodyweightwas notstatistically differentfromthe control mean. Nostatisticallysignificantdifferences infood consumptionbetween control andtreated animalswere detected withthe exception ofan increase inmeanfemalefoodconsumptioninthe 350 mg/kg/day group duringweeks -1, 1,and2 oftreatment. Meanfemalefood consumptionat that thosetimepointswere 18, 17and 17gversusl6, 15and 15g inthe control group, respectively.
Functional observation battery andmotoractivity resultsfromtreated animalswere comparedstatistically bysexandtreatmentdaytotheir respective control groups(pc 0.05). These parameterswere notaffected bytreatment withthe test article. Nostatisticallysignificantdifferences were observed betweentreated andcontrol animalsatanytime point.
Nostatistically significantdifferences betweentreated andcontrol animals werefoundfor hematologyparameterswiththe exception ofan increasein the meanactivated partialthromboplastintime inthe 1000mg/kg/day malesonweek4 and adecreaseinthe meanprothrombintimeinthe 1000 mg/kg/day femalesonweek4. Thesestatisticaldifferenceswere notof toxicological significance.
Notoxicologically significanteffects on serumchemistryvalues relatedto test article administrationwere observed atthe28-day primaryand42-day recovery sacrifice. Scattered instancesof statisticallysignificant differences betweentreated andcontrol animalswere detectedfor some serumchemistry parametersatthe28-day primarysacrifice. These scattered statisticaldifferenceswere not consideredtoxicologically significant becausethestatisticaldifferences occurred: inthe absenceofa doseresponse,inthe absenceof the accompanying clinicalchemistry changesexpected, inthe oppositedirectionfromwhatoccurs inatoxic state, ina directionwhich iswithout physiologicsignificance, ordueto potential interference withthe laboratory method. Nostatisticallysignificant differences inserumchemistry parameterswere detected between groups at the42-day recovery sacrifice. Nogrosslesionsthat could beattributedto thetest articleweredetected at either necropsy. Gross lesionswere nonspecific, lowinincidence, nondose-related andconsideredincidental.
No microscopiclesionsthat couldbeattributed to thetest articlewere detected onhistopathologicexam. Microscopicchangeswere nonspecific, low inincidence, non-dose-relatedandconsidered incidental.
Nostatisticalsignificantdifferences inorganweight ororganto bodyweight ratioswere detected betweencontrol andtreated animalswith one exception. Absolute liverweightswere statistically significantly increased with respectto control meansatthe28-day sacrifice inmalesinthe high dose andfemalesinthe midand highdose. Liver to bodyweight ratiosin midand highdosemalesandlow, midandhighdosefemaleswere statistically significantly increasedatthe28-day sacrifice. At the recovery 66188

## 5. Toxicity

Reliability Flag 29.12.2004 Type Species Sex Strain Route of admin. Exposure period Frequency of treatm. Post exposure period Doses Control group NOAEL Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

sacrifice, male absolute and liver to body weight ratio were statistically comparable to the control mean whereas female absolute liver weights and liver to body weight ratio were statistically significantly increased with respect to control mean. The difference inabsolute liver weight between control and treated females was less pronounced at the end of the recovery period, indicating the increase in liver weight was reversible in females as well as males. Inthe absence of test article related histopathologic and serum chemistry changes, increases in liver weight are considered an adaptive, rather than atoxic response, are not uncommon in the rat, and are most likely the result of microsomal induction.
In conclusion, no systemic toxicity was observed at any dose level. Based on the results of this study, the NOAEL (No Observed Adverse Effect Level) of HBCD administered orally to male and female rats for 26 consecutive days was 1,000 mglkglday. Chengelis C. 1996. A 26-day repeated dose oral toxicity study of HBCD in rats. Study No. WIL-186004. WIL Research Laboratories, Inc. Ashland,

: Eyvalid without restriction : Risk Assessment, Critical study for SIDS endpoint (32)

: Sub-chronic : rat : male/female : Sprague-Dawley : gavage : 90 Days : Once Daily : 28 Days : 0,100,300,1000 mg/kg/d

- yes, vehicle

. concurrent : = 1000

- ml/kg bw : EPA OPPTS 870.3100 : 2001 : yes : as prescribed by 1.1

- 1.4

: An Oral (Gavage) 90 Day Toxicity Study of HBCD in Rats. (Sponsor: ACC Brominated Flame Retardant Industry Panel)
This study was conducted according to OECD and GLP guidelines. The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc.
The test article, a composite ofthree lots of commercial hexabromocyclododecane (HBCD), was administered by oral gavage in corn oilonce daily to four groups of Crl:CD(SD)IGS BR rats (n=lS/sex/group) at dose levels of 0 (control), 100 (low), 300 (mid) and 1000 (high) mg/kg/day seven days per week for 90 days. The dosage volume was 5 ml/kg. The control animals received the vehicle, corn oil, only. At the end of the go-day treatment period, 10animals/sex/group were euthanized and necropsied. The remaining rats continued on test untreated for a28-day recovery period prior to necropsy. In addition to the main toxicology groups, two satellite groups of 20 animals/sex/group were treated concurrently inan identical manner at dose levels of 0 or 1000 mg HBCD/kg/day for up to 90 days. Body weights were recorded weekly. Two animals/sex/group were euthanized on study days 2, 6, 9, 13,20,27, 55, 89, 104 and 118, and blood and body fat 67188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

(mesenteric and/or omental) were collected. The body fat was analyzed for HBCD content. Animals inthe main toxicology groups were observed twice daily throughout the study for mortality and morbidity. Body weights and food consumption were measured weekly. Blood was collected at study weeks 3 (n=Vsex/group), 13 (n=l O/sex/group) and 17 (n=5/sex/group) for hematology, serum chemistry and hormone (T3, T4 and TSH) measurements. Urine was collected prior to each necropsy, at study weeks 13and 17, for urinalysis. Ocular examinations were performed prior to study initiation and during study weeks 12and 15. Functional Observational Battery and Locomotor Activity evaluations were performed on 5 animals/sex/group prior to study initiation, during the last week of test article administration (study week 13), and during the recovery period. An examination of vaginal cytology (for estrus cycle determinations) was performed on study days 69-90. At each necropsy, sperm motility/viability, morphology, and number were assessed. Complete necropsies were performed on allanimals. Approximately 40 organs or tissues/animal were collected and preserved. The adrenals, brain, epididymides, heart, kidneys, liver, ovaries, prostate, spleen, testes, thymus, thyroids with parathyroids, and uterus with cervix were weighed. Paraffin sections of tissues stained with hematoxylin and eosin from the control and 1000 mg/kg/day dose groups and the liver, lungs and thyroid glands inthe 100 and 300 mg/kg/day doses, and gross lesions from allanimals were examined under the light microscope. Livers from five randomly chosen animals/sex from the control and 1000 mglkglday dose groups were examined microscopically using Oil Red 0 or periodic acid Schiff’s (PAS) reagent for evidence of lipidacoumulation or glycogen accumulation/depletion, respectively. Statistical comparisons by sex and treatment day were made between the control and treated animals where indicated (~~0.05).
No test article-related effect on mortality occurred. Clinical signs were nonspecific, low in incidence, non-dose-related and not related to test article administration. No test article-related changes occurred in body weight, food consumption, Functional Observational Battery or Locomotor Activity. No test article-related effects on hematologic parameters were noted. No test article-related ocular lesions were detected at the ophthalmic exams. No test article-related changes were noted on the estrus cycle as determined by vaginal cytology, or on sperm motility/viability, morphology, and number. Instances of statistically significant differences between control and some treatment groups were detected at study week 13 inthe clinical chemistry data, hormone data, organ weight data and histology findings. They were considered secondary to hepatic enzyme induction or were otherwise not considered adverse effects of treatment as discussed further below. Statistically significant (~~0.05) test article-related serum clinical chemistry changes at week 13 include an increase inalbumin (all dose levels for males), total protein (all dose levels for females and 1000 mg/kg/day for males), globulin (300 and 1000 mg/kg/day for females), and chloride (all doses for both sexes). In addition, increased gamma glutamyltransferase levels were noted inthe 1000 mg/kg/day group (~~0.05). Thyroxine (T4) levels were decreased at study week 13 compared to the control mean in all male dose groups and the 300 and 1000 mg/kg/day dose females (~~0.05). There were no corresponding statistical effects on T3 and TSH.
While potentially test article-related, the changes inserum chemistry parameters were not of sufficient magnitude to be adverse, occurred in otherwise clinically normal animals, tended to be within or close to historical control values, and were not present atthe end of the recovery period. The increased serum albumin and gamma glutamyltransferase 68186I 

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

levels were considered related to the increased in liver weight intreated animals that was believed due to enzyme induction. Hepatic GGT is inducible inthe rat (Teschke et al 1983. 

### Gut. 24(7):625-30; Chandar and Nagarajan 1984. Biochem Int. Jan;8(1):41-8), and this induction has been shown to increase serum GGT levels inthe rat (Goldberg 1980. Crit Rev Clin Lab Sci. 12(1):1-58; Teschke et al. 1983. Gut 24(7):625-30; Nishmura and Teschke 1982. Biochem Pharmacol. Feb 1;31(3):377-81; Satoh et al. 1982. J Pharmacol Exp Ther. Jun;221(3):795-800). The increase inserum chloride was considered secondary to be presence of free bromide inthe test article preparation that interfered with the chloride determination methodology. The decrease inT4, which was also reversible, was considered related to the increased liver weights inthat induction of hepatic UDPG is known to enhance the elimination ofT4. The rat is particularly sensitive to hepatic enzyme induction.
The incidence of observations noted at gross necropsy was low and was not accompanied by evidence of frank organ damage. On histopathologic examination, mild findings were detected in both the control and treated groups. Potential test article?related histologic changes were identified in the liver and thyroid glands, but these were not considered to be indicative offrank toxicity. The liver changes in male rats at the go-day necropsy (Study Week 13) were characterized as minimal hepatocellular vacuolation and occurred in 10% of control males and -50% of the males at 100, 300 and 1000 mg/kg/day. Thus, there was no increase in incidence with dose. Minimal hepatocellular vacuolation was also detected infemales inthe control and treated groups without a clear dose response (3 to 4/l 0 animals per group). Mild and moderate vacuolation was detected in females only inthe 300 (l/l 0) and 1000 mg/kg/day (2/l 0) dose groups. Minimal to mild hepatocellular hypertrophy was also detected only inthe 1000 mg/kg/day group (5/10) females. Minimal thyroid follicular cell hypertrophy was detected l/10, l/10, 5/10 and 7/10 males inthe control, 100,300 and 1000 mg/kg/day groups, respectively and in4/l 0and 3/l 0 females inthe 300 and 1000 mg/kg/day groups respectively. In addition, mild thyroid follicular hypertrophy was detected in4/10 females inthe 1000 mg/kg/day group. The histologic changes inthe liver were accompanied by an increase in liver weight. In contrast there were no statistically significant changes in thyroid weight (absolute, relative to body weight and relative to brain weight). At study week 13, mean liver weights inalldose levels of both sexes (absolute, relative to body weight and relative to brain weight) were increased compared to the male and female control means (~~0.05).
The changes seen inthis study, an increase in liver weight, the lack of adverse histologic effects inthe liver and the apparent normal functioning ofthe liver, are consistent with enzyme induction (Amacher et al. 1998. Food Chem. Toxicol. 36:831-830), and hepatic enzyme induction is an adaptive, and not an adverse, effect (Williams and latropoulos 2002. Toxicol Pathol. Jan-Feb;30(1):41-53). Hepatocellular vacuolation and hypertrophy, seen inthis study, often accompany the increased liver weight 

### caused by liver enzyme induction, and were reversible. At week 17,the liver changes (weight and histology) had at least partially, if not fully, resolved inalltreated groups without delayed or long-tern toxic effects.
Limited pharmacokinetic studies indicate HBCD is extensively metabolized prior to excretion inthe feces and urine. The results of the fat analysis in this study indicate that mammalian system handles the 3 HBCD stereoisomers differently, and may be less efficient at eliminating one stereoisomer over another. (The relative isomer concentrations inadipose tissue at alltime points were alpha>>gamma>beta in contrast to the test article composition of gamma>>alpha>beta.) Thus, itshould not be unexpected that hepatic enzyme induction occurs with exposure to 69188

## 5. Toxicity

Reliability Flag 29.12.2004 Type Species Sex Strain Route of admin. Exposure period Frequency of treatm. Post exposure period Doses Control group NOAEL Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

substantial doses go-day study. over a significant period oftime, as was the case inthe

The histologic changes inthe thyroid consisted of a slight increase inthe incidence of minimal follicular cell hypertrophy inthe high dose males and minimal or mild hypertrophy inthe high dose females. Itwas not readily apparent that these minimal changes were an effect of treatment, and in any event appeared reversible. The follicular cell hypertrophy may have been related to serum T4 levels. Follicular cell hypertrophy is the normal physiological response to reduced to serum 14 levels and isthe typical adaptive response of a healthy normally functioning organism acting to maintain serum T4 levels inthe normal range. The mean prostate weight was increased inthe 1000 mglkglday group males at the primary necropsy. This was not considered to be of toxicological significance since the increase did not persist to the recovery period, there were no correlating histologic findings and no change in seminal quality.
HBCD was detected inthe adipose tissue of male and female rats treated with 1000 mglkglday for upto 90 days. Isomer-specific analysis showed that the relative isomer concentrations inadipose tissue at alltime points were alpha>>gamma>beta which is in contrast to the test article composition (gamma>>alpha>beta). Steady state levels were achieved by study day 27. Levels in male and female rats were similar at alltime points and declined during the recovery period.
Allthe test article-related changes at 100 and 300 mg/kg/day were mild, reversible, generally secondary to hepatic enzyme induction (which is an adaptive not atoxic change) and without effect on the clinical condition of the animals. The findings observed at 1000 mg/kg/day (increased serum gamma glutamyltransferase, increased liver and prostate weights), were also reversible, not associated with specific target organ damage or diminished function and were, therefore, of limited, ifany, toxicologic significance. On this basis, the no-observed-adverse-effect level (NOAEL) of HBCD administered to Crl:CD@(SD)IGS BR rats by gavage in corn oilfor 90 days is 1000 mg/kg/day. Chengelis C. An Oral (Gavage) 90 Day Toxicity Study of HBCD in Rats. Study No. WIL-186012. WIL Research Laboratories, Inc., Ashland, Ohio. 2001. (1) valid without restriction Risk Assessment, Critical study for SIDS endpoint (33)

.

. Sub-chronic : : :

. .

.

.

.

. : : : : rat male/female Sprague-Dawley oral feed 28 Days Daily None 0, 1,2.5, 5% Diet yes, concurrent no treatment =l- % : : :

. other 1969 no data other TS: Hexabromid S (HBCD formerly manufactured by BASE)

. 28-Day Feeding Study of HBCD in Rats. (Sponsor: BASF) 70188

## 5. Toxicity

Reliability 29.12.2004 Type Species Sex Strain Route of admin. Exposure period Frequency of treatm. Post exposure period Doses Control group NOAEL Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

HBCD (“Hexabromid s”) was tested in Sprague-Dawley rats (IO/sex/group) at doses of 0, 1,2.5 and 5% ofthe diet for 28 days. Doses calculated from the actual body weights and food consumption inthis study are 0, 940, 2410, and 4820 mg/kg body weight/day.
No clinical signs related to treatment were observed atthe 1% dose level. Body weights at the 1and 2.5% dose levels were comparable tothe controls. Liver weights (absolute and relative to body weight) were increased at alldose levels, but no microscopic pathology was detected. Thyroid hyperplasia was observed insome animals at alldoses, and “very slight numerical development of the follicles and ripening follicles inthe ovaries of females” at the high dose (4820 mg/kg/d) was reported. No changes inany other organ related to treatment and no changes inclinical chemistry tests were detected.
The report concluded that “The increased liver weight must be attributed to hyperactivity; hypermetabolism as a result of increased thyroid activity appears probable inview of the observations of the thyroid’. Therefore, the increased liver weights were not pathologic: there were no microscopic lesions detected on histopathology and no change in clinical chemistry values.
Zeller H and Kirsch P (1969) Hexabromocyclododecane: 28-day feeding trials with rats. BASF Unpublished Laboratory Report. : (2) valid with restrictions

. Sub-chronic

.

. rat

. . male/female

. Sprague-Dawley

.

. oral feed : 90 Days

.

. Daily : None

. . 0, 0.16, 0.32, 0.64 and 1.28% Diet

.

. yes, concurrent no treatment

.

. = 1.28- % : other : 1970 : no data

. other TS: Hexabromid S (HBCD formerly manufactured by BASF)

. 90-Day Feeding Study of HBCD in Rats (Sponsor: BASF)
HBCD (“Hexabromid S”) was tested in Sprague-Dawley rats at doses of 0, 0.16, 0.32, 0.64 and 1.28% of the diet for 90 days. Doses calculated on the actual body weights and food consumption inthis study reveals: 0, 120, 240,470 and 950 mg/kg body weight/day.
Doses up to 0.64% (470 mg/kg/d) produced no adverse clinical signs, no change in body weight, and no change inclinical chemistry results. An increase inthe relative liver to body weight ratio was found, and was accompanied by fatty accumulation but no other histologically discernible changes were detected inthe liver. Further, no histological changes were found inany other organ. The original report stated that inthe “absence of detectable clinico-chemical disturbances or histological changes of the vital organs, itwas concluded that the increased liver weight and the fat deposits, both of which were largely reversible when administration of Hexabromid S was stopped, were the result of a temporary increase inthe 71 I88

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

activity of the liver.” Thus, no adverse effect was produced at the highest dose tested, 1.26% ofthe diet.
Zeller H and Kirsch P (1970) Hexabromocyclododecane: 90-day feeding trials with rats. BASF Unpublished Laboratory Report. Reliability : (2) valid with restrictions 29.12.2004 (35)

5.5 GENETlC TOXlCfTY ‘IN VITRO‘

Type : Ames test System of testing : Salmonella microsomal Test concentration : Various Cycotoxic concentr. : Metabolic activation : with and without Result : negative Method : other Year

. . GLP : no data Test substance : other TS
Attached document : HBCD has been tested for mutagenicity inthe Ames Salmonella microsomal assay, both with and without metabolic activation, in multiple tests. All results were negative: Ogaswara S and Hanafusa T. (1993) Report on mutagenicity test on Pyroguard SR-103 using microorganisms;
Baskin A and Phillips, B. (1977) Mutagenicity oftwo lots of FM-100, Lot 53 and residue of Lot 3322 inthe absence and presence of metabolic activation. Industrial Biotest Laboratories, Sponsored by Velsicol Chemical Corporation; Anonymous. (1979) Mutagenicity test of GLS-S6-41A. Gulf South Research Institute, Sponsored by Ethyl Corporation;
US Environmental Protection Agency (1990) Ames metabolic activation test to assess the potential mutagenic effect of Compound No. 49. Letter from BASF. EPA/OTS Dot #86-900000385; Simmons V., Poole, D., Newell, G., and Skinner, W. (1976) Invitro microbiological mutagenicity studies for four CIBA-GEIGY Corporation compounds. SRI Project LSC-5702. Reliability : (1) valid without restriction Flag : Risk Assessment, Critical study for SIDS endpoint 28.12.2004 Type : Chromosomal aberration test System of testing : human peripheral blood lymphocytes Test concentration : 10,19,38,75,150,300 and 600 ug/ml Cycotoxic concentr. : 750 @ml Metabolic activation : with and without Result : negative Method : EPA OPPTS 870.5375 Year : 1996 GLP : yes Test substance : as prescribed by 1.1

- 1.4
Attached document : Invitro Chromosomal Aberration Test. (Sponsor: ACC Brominated Flame Retardant Industry Panel) 72188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

’ The test article was a composite of 3 lots of commercial HBCD produced

by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current EPA, OECD and GLP guidelines.
HBCD was tested inthe invitro mammalian cytogenetic test using human peripheral blood lymphocytes both inthe absence and presence of metabolic activation. The assay was performed intwo phases. The first phase, the initial chromosome aberration assay, was conducted to establish the dose range for testing and to evaluate the clastogenic potential ofthe test article. The second phase, the independent repeat chromosome aberration assay, was performed to confirm the test system response to the test article seen inthe initial assay. Dimethylsulfoxide was used as asolvent. Inthe initial assay, the maximum dose tested was 2,500 @ml. Dose levels greater than 2,500 ug/ml were insoluble intreatment medium. Visible precipitate was observed in treatment medium at 750 and 2,500 ug/ml and was soluble but cloudy at dose levels of 75 and 250 ug/ml. The test article was soluble intreatment medium at allother doses tested. Inthe non-activated portion of the initial assay cells were exposed to the test article continuously for 20 hours; in the SS-activated portion ofthe initial chromosome aberration assay, cells were exposed to the test article for 4 hrs. Metaphase cells were collected at 20 hrs after initiation of treatment. Dose levels of 2,500 q/ml inthe nonactivate study and 750 and 2,500 ug/ml inthe SS-activated study were not analyzed for chromosome aberrations due to complete mitotic inhibition. Toxicity (mitotic inhibition) of -56% was observed at the highest dose level (750 ug/ml) evaluated for chromosome aberrations, inthe non-activated study. Inthe SS-activated study, 13% toxicity was observed at the highest dose level (250 @ml) evaluated for chromosome aberrations. No statistically significant increases in chromosome aberrations were observed in either the non-activated or SS-activated test systems relative to the solvent control group regardless of dose level.
Based on the results of the initial assay, an independent repeat chromosome aberration assay was conducted inthe absence and presence of an Arochlor-induced S9 metabolic activation system at dose levels of 10, 19, 38, 75, 150, 300 and 600 t&j/ml. The test article was soluble but cloudy at 75 ug/ml and was workable intreatment medium at dose levels 150 ug/ml and higher. The test article was soluble intreatment medium at allother concentrations tested. Inthe independent repeat assay, cells were exposed to the test article continuously for 20 or 44 hr in the non-activated test system and for 4 hours inthe SS-activated test system. Metaphase cells were collected for microscopic evaluation in both the non-activated and SS-activated studies at 20 and 44 hrs after initiation oftreatment. Toxicity, measured by mitotic inhibition, was -55% and 94% atthe 20 and 44 hr harvests, respectively, at the highest dose levels (600 and 300 @ml) evaluated for chromosome aberrations inthe nonactivated studies. Inthe SS-activated studies, toxicity was approximately 71% and 69% at the 20 and 44 hr harvests, respectively, at the highest dose levels (600 and 300 us/ml) evaluated for chromosome aberrations. The 600 q/ml dose level inthe non-activated 44 hr harvest and inthe SS-activated 20 hr harvest was not analyzed for chromosome aberrations due to an insufficient number of storable metaphase cells. No statistically significant increases instructural chromosome aberrations were observed in either the non-activated or SS-activated studies, regardless of dose level or harvest time. No statistically significant increases in numerical chromosome aberrations were observed in either the non-activated or S9- activated studies at the 44 hr harvest time, regardless of dose level. HBCD was negative for the induction of structural and numerical chromosome aberrations in human peripheral blood lymphocytes. 73188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

Gudi, Ft.and Schadly, E. 1996. Laboratory Study Number G96A061.342. Microbiological Associates, Inc., Rockville, MD. Reliability (1) valid without restriction Flag Risk Assessment, Critical study for SIDS endpoint 29.12.2004 (36) Type : other System of testing

. . Hamster cells Test concentration : see freetext Cycotoxic concentr.

. . Metabolic activation

. . Result

. . Method : other Year : 1999 GLP : no Test substance

. . no data
Attached document

. . In Vitro latrogenic Recombination
The Sp5 and SPD8 cell lines were developed by the paper’s authors. The clones used inthis study exhibit aspontaneous partial duplication of the hprt gene, resulting ina non-functional hgprt protein. These mutants revert spontaneously to afunctional hprt gene phenotype by recombination with a frequency of 1x 105 reversions/cell generation. This reversion frequency is said to increase by exposure to chemical or physical agents. Treatment with the test substance was for 24 hr at37 degrees C. HBCD was tested invitro in hamster cells (Sp5N79 and SPD8) ina recombination assay at five doses between 2 and 20 @ml plus a control. Inthe SPD8 cells, HBCD concentrations of 0, 3,6, 10, 15, and 20 &j/ml resulted ina reversion frequency of 1.O,0.7, 0.8, 0.9, 1.4, and 1.9, respectively. Cytoxicity was observed atthe 20 @ml dose. Inthe Sp5 cells, HBCD concentrations of 0, 2, 5, 10, 15, 20 @ml resulted ina reversion frequency of 1.0, 1.0, 0.8, 1.1, 1.4 and 2.2, respectively. Cytotoxicity was not observed. The reversion frequency at the 20 @ml dose for the Sp5 and SPD8 cells was statistically different from the control (Student’s ttest, ~~0.05). Treatment with HBCD resulted inan - maximal 2-fold increase in revertant frequency. (Helleday et al. Brominated flame retardants induce intragenic recombination in mammalian cells. Mutation Research 439 (f999) 137-l 47). This is a non-standard genetic toxicity test, and its reliability and predictive ability is unknown. This is not atest used by regulatory agencies to assess genotoxicity potential. Reliability : (4) not assignable 20.12.2004 (37)

5.6 GENETlC TOXICITY ‘IN VIVO



### Type

. . Micronucleus assay Species

. . mouse Sex : male Strain : Route of admin. : i.p. Exposure period Once Doses : 0,500, 1,000 or 2,000 mg/kg bw Result : negative Method : OECD Guide-line 474 “Genetic Toxicology: Micronucleus Test” Year : 2000 GLP : yes Test substance : as prescribed by 1.l

- 1.4 74188

## 5. Toxicity Id Date 25637-99-4 31.03.2005

Attached document : In Vivo Mouse Micronucleus Test (Sponosor: BASF Corporation)
The test article was a composite of 3 lois of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. This study was performed according to current OECD guidelines and Good Laboratory Practices.
HBCD dose levels administered intraperitoneally to male mice were 0, 500, 1,000 or 2,000 mg/kg body weight. The negative control animals were administered thevehicle, DMSO. Cyclophosphamide and vincristine were used as positive controls and responded as expected. HBCD-treatment did not increase in number of polychromatic erythrqcytes containing either small or large micronuclei. Micronuclei formation in HBCD-treated mice was within the same range as that of the concurrent negative control and within the range of historical control data. No evidence of chromosome damaging (clastogenic) effects was observed. There was no indication of any impairment of chromosome distribution inthe course of mitosis. HBCD was clearly negative for clastogenicity and the ability to induce spindle poison effects inthis mouse micronucleus test.

Reliability Flag 29.12.2004 : : Engelhardt, G and Hoffmann, H. (2000) Laboratory Project Identification: 26M0100/004018. Experimental Toxicology and Ecology, BASF Aktiengesellschaft, Ludwigshafen, Germany. (1) valid without restriction Risk Assessment, Critical study for SIDS endpoint (38)

5.7 CARGINOGENICITY
Species Sex Strain Route of admin. Exposure perlod Frequency of treatm. Post exposure period Doses Result Control group Method Year GLP Test substance :

. . : : :

.

. .

. : : : :

. .

.

. mouse male/female B6C3Fl oral feed 18 months daily none 0, 100, 1000 or negative yes, concurrent other

no data other TS 10,000 ppm

no treatment

Attached document .

. Male and female mice were 10,000 ppm for 18 months. any dose level. This study Toxicology, National Public Test and Research Center, the study is available. fed diets containing HBCD at 0, 100, 1000 or There was no evidence of carcinogenicity at was performed bythe Department of Health Research Institute, Biologjcal Safety Japan (date not specified). Only a summary of

The test Seiyake substance was the commercial HBCD product from Daiichi Kogyo K.K. There were 50 males and 50 females per dose level.
There was no effect of test article on mortality, clinical signs, body weights or food consumption. A variety of gross lesions/nodules were detected at necropsy which were not correlated with administration of test article. The 75188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

main histopathologic change noted was hepatocyte swlling, degenration, ncerosis, vacuole formation and fatty infiltration, and were suspected to be related to hepatic enzyme induction. There was no correlation of these changes with dose level. Various tumors were observed in many organs, but such incidence was sporadic and not due to test article administration. Reliability : (2) valid with restrictions 28.12.2004 (39)

5.8.1 TOXlClTY TO FERTlLlTY

5.8.2 DEVELOPMENTAL
Species Sex Strain Route of admin. Exposure period Frequency of treatm. Duration of test Doses Control group NOAEL maternal tox. NOAEL teratogen. Result Method Year GLP Test substance
Attached document TOXICIN/TERATOGENlClN

.

. rat

.

. female : Sprague-Dawley

.

. gavage : Day 6-19 of Gestation : Once daily, 7days/wk : : 0,250,500,1000 mg/kg/d : yes, concurrent vehicle : >= lOOO- mg/kg bw

. . a= lOOO- mg/kg bw

.

. Not a developmental toxicant : EPA OPPTS 870.3700

. 1999

. : yes : as prescribed by 1.l

- 1.4

: A Prenatal Developmental Toxicity Study of Hexabromocyclododecane (HBCD) in Rats. (Sponsor: ACC Brominated Flame Retardant Industry Panel)
The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. The study was performed according to EPA, OECD and GLP guidelines. This study was required by KEMI (without consultation of the EU Technical Meeting) because KEMI decided the existing study inthe literature (Murai et al.) was insufficient.
HBCD was administered in corn oil by gavage to 25 presumed pregnant Crl:CD(SD)IGS Br rats/group once daily from gestation days 6-19 at doses of 0, 250, 500 or 1,000 mg/kg/day. Control animals received corn oilonly. Female rats were mated in-house and were treated daily on gestation days 6-19 with HBCD via gavage at dose levels of 0 (vehicle control), 250, 500 or 1000 mg/kg/day at a constant volume of 5 ml/kg. Individual doses were based on the most recent body weight. The day on which evidence of mating was observed was considered day 0 of gestation. Dams were observed daily and maternal body weight and food consumption measured at appropriate intervals. Females were euthanized on day 20 of gestation and necropsied. Gravid uterine and liver weights were recorded. Liiers were delivered by cesarean section. The total number of corpora lutea, total number of implantations, early and late resorptions, number and location of allfetuses, and the sex and individual weights offetuses were recorded. All fetuses were examined grossly. Approximately one-half of the fetuses in each litter were stained with Alizarin Red S and Alcian Blue and evaluated for skeletal/cartilaginous malformations and ossification variations. The maternal day 20 gestation examinations and cesarean
76 I88

## 5. Toxicity

Reliability Flag 29.12.2004 Species Sex Strain Route of admin. Exposure period Frequency of treatm. Duration of test Doses Control group NdAEL maternal tox. NOAEL teratogen. Result Method Year GLP Test substance
Attached document Id 25637-99-4 Date 31.03.2005

sections, and subsequent fetal evaluations were performed blind to treatment. No mortality occurred during the course of the study. No treatment-related clinical signs were observed. Body weight gain and food consumption were not adversely affected. No treatment-related findings were detected at necropsy. Intrauterine growth and survival were unaffected by treatment. No treatment-related fetal malformations or developmental variations were observed. The no-effect level (NOEL) for maternal toxicity and developmental toxicity was 1,000 mg/kg/day, the highest dose tested Stump, D. 1999. A Prenatal Developmental Toxicity Study of Hexabromocyclododecane (HBCD) in Rats. Laboratory Study No.: WIL186009. WIL Research Laboratories, Inc., Ashland, OH. : (1) valid without restriction : Risk Assessment, Critical study for SIDS

. . rat : female

.

. other

.

. oral feed : Day O-20 of Gestation : Daily

.

. Day O-20 of Gestation or Day 0 Gestation endpoint (40)

through 7 wks postpartum : 0, 0.01, 0.1, or 1% of diet

. . yes, concurrent no treatment

. . >= 1 - %

.

. >= 1 - % : Not adevelopmental

.

. other

.

. 1985

. . no data

.

. other TS toxicant

: Murai et al. 1985 (Pharmacometrics (Japan) 29(6):981-986) identified no reproductive or developmental effects inthe rat at doses up to 1% inthe diet administered from days O-20 of gestation. This dose is approximately equivalent to 500 mg/kg/d.
The Murai et alstudy consisted of a7 day dose range finding study (n=5 rats/dose group) and a combined teratogenicity-developmental study (n=20/dose group). Doses inthe 7 day range finding study were 0, 0.3, 1, 3 or 10 g/kg/day. Doses as high as 10 g/kg/day produced no evidence of toxicity. A statistically significant (PcO.01) increase in liver weight was noted in groups receiving > 1g/kg/day. Doses for the combined teratogenicity-developmental study were based on this increase in liver weight. In the combined teratogenicity-developmental study, pregnant female rats were fed diets containing 0, 0.01, 0.1, or 1% HBCD on days O20 of gestation. Daily doses were estimated by the authors to be 0, 5, 50 or 500 mg/kg/day and the average total dose/rat/group was estimated to be 0, 0.13, 1.28 or 12.0 g/kg. Rats were observed daily and body weight and food consumption measured. Fourteen rats from each group were sacrificed on day 20 of gestation and their fetuses were examined for toxicity.or teratogenicity. Approximately 150 fetuses/dose level were examined for evidence of teratogenicity. All fetuses from all litters were examined for signs of external anomalies. Approximately 2/3 of the fetuses/dam were examined for skeletal abnormalities; the remaining fetuses from each dam were examined for any abnormalities ofthe internal organs. In addition, six rats from each group were allowed to deliver their litters and growth of the litters was observed until the 7th week postparturition. 77188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

The authors’ estimated the doses inthe feed were equivalent to 0, 5,50 or 500 mg HBCD /kg body weight /day. No adverse effects were detected in any treatment group with respect to maternal weight gain, food consumption, or gross appearance of internal organs. The mean liver (absolute and relative to body weight) weight inthe 1% group was statistically different (higher) from the control mean. Normal development was seen in neonates carried through to six weeks of age. There was no adverse effect of treatment on the number of corpora lutea, implants, resporptions, livefetuses, sex ratio, or body or placental weight. No fetal deaths occurred in any group. No external, skeletal or visceral malformations were detected. Afew skeletal variations were detected but where of similar types and numbers inthe control and treated groups. There was no significant differences between the control and treated groups inthe number of implantation, live newborns, dead newborns, newborn parturition index. The weaning and survival index was comparable inthe control and treated groups. Body weight changes newborns was comparable inall groups. live
inthe

No reproductive or developmental effects where detected in rats at HBCD doses up to 1% inthe diet (-500 mglkgld) administered from days O-20 of gestation. Further, normal development was seen in neonates carried through to six weeks of age. Dose levels: 0, 0.01, 0.1, or 1% HBCD on days O-20 of gestation [Murai estimate: 0, 5, 50 or 500 mg/kg/day]. No teratogenic effects. Normal development in neonates carried through age 6wks. NOEL = 1% of diet.

Reliability 28.12.2004 : Murai, T. Kawasaki, H., Kanoh, S. 1985. Studies on the toxicity of insecticides and food additives inpregnant rats -fetal toxicity of Hexabromocyclododecane. Pharmacometrics (Japan) 29(6):981-986). (1) valid without restriction (41)

14.12.2004

14.12.2004

5.8.3 TOXICITY TO REPRODUCTION, OTHER STUDIES



### We : other In vitro/in vivo : Invivo Species : rat Sex : male/female Strain : Route of admin. Exposure period Frequency of treatm. Duration of test Doses Control group : Result : See Freetext
Attached document : Two teratology studies on HBCD are available; one published inthe literature (Murai et al. 1988; high dose = 1% of the diet) and one recently completed by industry (1999) under current guidelines and Good 

# I 78188 I

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

Laboratory Practices using the HBCD in commercial production and use (high dose = 1,000 mglkgld). Both studies are negative for developmental toxicity. Repeated dose studies (two 28 day studies, one 90 day study, and one 18 month study inasecond species) indicate HBCD does not affect the reproductive organs at doses up to 1,000 mg/kg/day. According to the OECD SIDS Manual, when teratology and 90 day studies show no effects on the reproductive system then the requirement for the reproductive endpoint are met. Teratology, 28 day, 90 day and 18 month studies alldemonstrate HBCD has no effect on the reproductive system at the limit dose of 1,000 mglkgld.
Inthe 2001 90-Day study, considered the most informative ofthe subchronic studies mentioned inthe preceding paragraph, the following organs of the reproductive tract were weighed at the go-day and recovery sacrifices: epididymides (total and cauda), ovaries (with oviducts), prostate, testes, and uterus and cervix. These organs plus the semininal vesicles, vagina and vas deferens were examined histologically inanimals inthe control and high dose groups. Vaginal smears for determination ofthe stage of estrus were obtained form allfemales once daily beginning study day 69 through the go-day necropsy. An assessment of spermatogenesis was performed at the go-day sacrifice by evaluating sperm motility/viability, morphology and epidiymal and testicular sperm numbers and production rate.
No adverse effect on the estrous cycle was detected infemales receiving doses as high as 1,000 mg/kg/d for 90 consecutive days. No adverse effect on spermeogenesis was detected in males receiving doses as high as 1,000 mg/kg for 90 days. No test-article related changes inweight or microscopic effects were noted inthe organs of the reproductive tract with the sole exception of an increase inweight ofthe prostate at 1,000 mg/kg/d on day 90. Mean prostate weight was increased inthe 1,000 mglkgld dose group compared to the control mean after 90 days oftreatment (0.95, 0.99, 1.12, 1.25* g inthe control, 100, 300, and 1,000 mg/kg/d, respectively). The mean prostate weight relative to mean body and brain weights was also statistically increased at the high dose on day 90. At the recovery sacrifice, the prostate weights inthe control and high dose groups were not statistically different. No test article-related changes inthe prostate were detected on microscopic exam at either sacrifice. 28.12.2004

5.9 SPECIFIC INVESTlGATlONS

Endpolnt Neurotoxicity Study descr. in chapter 5.4 Repeated Dose Toxicity Reference Chengelis C. An Oral (Gavage) 90 Day Toxicity Study of HBCD in Rats. Study No. WIL-186012. WIL Research Laboratories, Inc., Ashland, Ohio. 2001 Type Species .

.

.

. other: rat Functional Observational Battery, Motor Activity Sex
. . male/female Strain . Sprague-Dawley Route of admin. Oral, gavage No. of animals : 10 Vehicle : other: corn oil Exposure period : 90 day(s) Frequency of treatm. : once daily Doses : 0,100,300,1000 mg/kg bw Control group : yes, concurrent vehicle Observation period : Result : negative 79188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

Method Year GLP Test substance
Attached document

Reliability Flag 15.12.2004 Endpoint Study descr. in chapter Reference 

## Type Species Sex Strain Route of admin. No. of animals Vehicle Exposure period Frequency of treatm. Doses Control group Observation period Result Method Year GLP Test substance
Attached document other: OPPTS, OECD 2001 Yes as prescribed by 1.l

- 1.4

.

. This study was conducted according to OECD and GLP guidelines. The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc.
The test article, a composite of three lots of commercial hexabromocyclododecane (HBCD), was administered by oral gavage in corn oilonce daily to four groups of Crl:CD(SD)IGS BR rats (n=15/sex/group) at dose levels of 0 (control), 100 (low), 300 (mid) and 1000 (high) mg/kg/day seven days per week for 90 days. The dosage volume was 5 ml/kg. The control animals received the vehicle, corn oil, only. At the end of the go-day treatment period, 10 animals/sex/group were euthanized and necropsied. The remaining rats continued on test untreated for a 28-day recovery period prior to necropsy. Functional Observational Battery and Locomotor Activity evaluations were performed on 5 animals/sex/group prior to study initiation, during the last week of test article administration (study week 13), and during the recovery period. No test article-related changes occurred inthe Functional Observational Battery or Locomotor Activity evaluations at any time point.
The NOELs for the Functional Observation Battery and the Motor Activity tests was 1000 mglkg bw, the highest dose tested.

.

. (1) valid without restriction

.

. Critical study for SIDS endpoint
Neurotoxicity 5.4 Repeated Dose Toxicity Chengelis C. 1996 A 28-day rats. Study No. WIL-186004. OH). : other: Functional Observational

.

. rat (42)

repeated dose oral toxicity study of HBCD in WIL Research Laboratories, Inc. Ashland,

Battery, Motor Activity

: male/female : Sprague-Dawley : oral, gavage : 24

.

. other: corn oil

. . 28 day(s)

. once daily

.

. 0,125,350,1000

. yes, concurrent

.

.

.

. negative : other: OECD : 1998

. Yes : as prescribed mg/kg bw vehicle

by 1.1

- 1.4

: This study was conducted according to OECD and GLP guidelines. The test article was a composite of 3 lots of commercial HBCD produced by Albemarle Corporation, Great Lakes Chemical Corporation, and Ameribrom, Inc. HBCD, inthe vehicle corn oil, was administered orally by gavage to three groups of Sprague-Dawley Crl: CD BR rats for a period of 28 consecutive 80188

## 5. Toxicity Id 25637-99-4 Date 31.03.2005

days. Dose levels were 125 (low), 350 (mid), or 1,000 (high) mg/kg/day, administered at dosage volume of 5 ml/kg. The test groups consisted of 6 males and 6females inthe 125 and 350 mg/kg/day groups and 12 males and 12females inthe 1000 mg/kg/day group. A concurrent control group comprised of 12 males and 12 females received the vehicle, corn oil,for 28 consecutive days at a dosage volume of 5 ml/kg. At the end of the dosing period, 6 animals/sex/group were sacrificed and necropsied. The remaining 6animals/sex inthe control and 1000 mg/kg/day groups remained on-test untreated for a 14-day recovery period. At the end of the recovery period, allanimals were sacrificed and necropsied. Functional observational battery and motor activity evaluations were performed during weeks -1 (pretest), 3, and 5 (recovery). Functional observation battery and motor activity results from treated animals were compared statistically by sex and treatment day to their respective control groups (p < 0.05). These parameters were not affected by treatment with the test article. No statistically significant differences were observed between treated and control animals at any time point.
The NOELs inthe Functional Observation Battery and the Motor Activity tests were 1000 mg/kg bw, the highest dose tested. Reliability : (1) valid without restriction Flag : Critical study for SIDS endpoint 15.12.2004 (43)

15.12.2004

5.tO EXPOSURE EXPERIENCE

5.11 ADDlTlONAL REMARKS

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## 6. Analyt. Meth. for Detection and Identification Id 25637-99-4 Date 31.03.2005

6.1 ANALYTlCAL METHODS

Test substance : HBCD Method : LCMS
Attached document : HBCD is susceptable to decomposition at temperatures > approximately 269 degrees Centigrade. TGA analysis of the commercial HBCD product indicates 50% weight loss at 269 degrees Centigrade and 90% weight loss at 274 degrees Centigrade. Thus, it is not ideally suited to gas chromatography because of the temperatures commonly employed inthat methodology. Also, a gas chromatography method that is able to separate the three stereoisomers has not been identified. As a consequence, the preferred analytical method is liquid chromatography coupled with mass spectrometry. An LCMS method has been developed by Wildlife International LTD under the sponsorship of the ACC Brominated Flame Retardant Industry Panel that can separate and quantitate the three HBCD stereoisomers invarious matrixes. (See the studies sponsored by BFRIP in2003 and 2004.) Reliability : (1) valid without restriction 29.12.2004

6.2 DETECTION AND IDENTlFICAT’lON

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## 7. Eff. Against Target Org. and Intended Uses Id 25637-99-4 Date 31.03.2005

7.1 FUNCTION

7.2 EFFECTS ON ORGANISMS TO BE CONTROLLED

7.3 ORGANISMS TO BE PROTECTED

7.4 USER

7.5 RESISTANCE

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## 8. Meas. Net. to Prot. Man, Animals, Environment Id 25637-99-4 Date 31.03.2005

8.1 METHODS HANDLING AND STORING

8.2 FIRE GUIDANCE

8.3 EMERGENCY MEASURES

8.4 POSSIB. OF RENDERING SUBST. HARMLESS

8.5 WASTE MANAGEMENT

8.6 SIDE-EFFECTS DETECTlON

8.7 SUBSTANCE REGISTERED AS DANGEROUS FOR GROUND WATER

REACTlVlTY TOWARDS CONTAlNER MATERIAL

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## 9. References Id 25637-99-4 Date 31.03.2005



### (1) Albemarle Corporation. Saytex HP-900 Flame Retardant. Technical Data Sheet. BC1033 (1l/00). (2) MacGregor J and Nixon W. 1997. Hexabromocyclododecane (HBCD): Determination of noctanol/water partition coefficient. Wildlife International LTD. Project Number: 4396-104. Wildlife International LTD. Easton, MD. (3) J. Stenzel and B. Markley. Hexabromocyclododecane (HBCD): determination of the water solubility. Wildlife Internation LTD. Project Number: 439C105. Wildlife international LTD. Easton, MD. (4) MacGregor J and Nixon W. 2004. Final Report. Determination of Hexabromocyclododecane (HBCD) Diastereomers using a Generator Column Method. Wildlife International Project Number: 439C138. Wildlife International, Ltd. Easton, MD.
Davis J, Gonsior Sand Marty G. 2003. Evaluation of Aerobic And Anaerobic Transformation Of Hexabromocyclododecane In Soil. Study Number 021082. Environmental Chemistry Research Laboratory. Toxicology & Environmental Research and Consulting. The Dow Chemical Company. Midland, Ml. (6) Schaefer E and Siddiqui A. 2003. Hexabromocylcododecane (HBCD): An Activated Sludge, Respiration Inhibition Test. Project Number: 439E-108A. Wildlife International, Ltd. Easton, MD. (7) Schaefer, Eand Haberlein, D., 1996, Hexabromocyclododecane (HBCD): Closed Bottle Test. Project No.: 439E-102. Wildlife International Ltd. Easton, MD. (8) Davis J, Gonsior Sand Marty G. Evaluation Of Aerobic And Anaerobic Transformation Of Hexabromocyclododecane InAquatic Sediment Systems. Study Number 021081. Environmental Chemistry Research Laboratory, Toxicology & Environmental Research and Consulting. The Dow Chemical Company Midland, Michigan. 2003. 

### (9) Davis JW, Gonsior SJ, Markham DA, and Marty GT. 2004. Investigation of the biodegradation of [14C]hexabromocyclododecane insludge, sediment, and soil. Laboratory Project Study ID 031178. Toxicology & Environmental Research and Consulting. The Dow Chemical Company, Midland, Ml. 

### (10) Drottar K. and Krueger H. 2000. Hexabromocyclododecane (HBCD): Flow-through bioconcentration test with rainbow trout (Oncorhynchus mykiss). Project No.: 439A-111. Wildlife International, Ltd. Easton, MD. 

### (11) Aufterheide et al. 2002. Effect of Hexabromocyclododecane on the Survival and Reproduction of the Earthworm, Eisenia fetida. ABC Study No. 47222. ABC Laboratories, Inc., Columbia, Missouri; Wildlife International, Inc., Easton, MD. (12) Graves, W and Swigert, J. (1997) Hexabromocyclododecane (HBCD): A96-Hour FlowThrough Acute Toxicity Test with the Rainbow Trout (Oncorhynchus mykiss). Project Number: 439A-101. Wildlife International LTD, Easton, MD. (13) Drottar et al. 2001. Hexabromocyclododecane (HBCD): An early life-stage toxicity test with the rainbow trout (Onchorhynchus mykiss). Project No.: 439A-112. Wildlife International, Ltd. Easton, MD. (14) Graves W and Swigert J. (1997) Hexabromocyclododecane (HBCD): a48-hour flowthrough acute toxicity test with the cladoceren (Daphnia magna). Project Number: 439A102. Wildlife International Ltd., Easton, MD. (15) Roberts C. and Swigert J. Hexabromocyclododecane (HBCD): A 96-Hour Toxicity Test with the Freashwater Alga (Selenastrum capricornutum). Wildlife International Ltd. Project Number: 439A-103. June 3, 1997. Wildlife International Ltd., Easton, MD.
85 I 88

## 9. References

(16)

(17) (18)

(19)

(20)

(21)

(22)

(23)

(24)
(25)

(26)
(27)

(28)
(2%
(30) (31) (32)
(33) Id 25637-99-4 Date 31.03.2005

Desjardins D, MacGregor J, Krueger H. 2004. Hexabromocyclododecane (HBCD): a 72- hour toxicity test with the marine diatom (Skeletonema costatum). Final Report. Wildlife International LTD Project Number: 439A-125. Wildlife International LTD, Easton, MD.
Walsh et al. 1987. Ecotoxicology and Environmental Safety, 14, 215-222.
Drottar K. and Krueger H. 1998. Hexabromocyclododecane (HBCD): Flow-through lifecycle toxicity test with the cladocerna (Daphnia magna). Project No.: 439A-108. Wildlife International, Ltd. Easton, MD.
Thomas J et al. 2003. Hexabromocyclododecane (HBCD): A Prolonged Sediment Toxicity Test with Hyalella azteca Using Spiked Sediment with 5% Total Organic Carbon. Project Number: 439A-120. Wildlife International, Ltd. Easton, MD.
Thomas S, Krueger H and Kendall T. 2003. Hexabromocyclododecane (HBCD): A Prolonged Sediment Toxicity Test with Hyalella azteca Using Spiked Sediment with 2% Total Organic Carbon. Project Number: 439A-119B. Wildlife International, Ltd. Easton, MD.
Porch et al. 2002. Hexabromocyclododecane (HBCD): A Toxicity Test to Determine the Effects ofthe Test Substance on Seedling Emergence of Six Species of Plants. Project Number: 439-103. Wildlife International, Ltd. Easton, MD.

Aufterheide et al; 2002. Effect of Hexabromocyclododecane on the Survival and Reproduction of the Earthworm, Eisenia fetida. ABC Study No. 47222. ABC Laboratories, inc., Columbia, Missouri; Wildlife International, Inc., Easton, MD. Toxicological Risks of Selected Flame Retardants”, National Academy Press, Washington, DC, 2000, pages 54-55
Ft.Arita, K. Miyazaki and S. Mure. 1983. Metabolic test of hexabromocyclododecane. Department of Pharmacy, Hokkaido University Hospital. Japan.
C.C. Yu and Y. H. Atallah. Velsicol Chemical Corporation, Laboratory Report. September 17, 1980.
Lewis C. 1978. Experiment Reference No. 78385-2 and 78385-l. Consumer Product Testing, Fairfield, NJ
Woolhiser M and Anderson P. 2003. Hexabromocyclododecane: Contact sensistization potential via the local lymph node assay (including primary irritancy screen) using CBA/J mice. Study ID 031013. Toxicology & Envionmental Research and Consulting. The Dow Chemical Company. Midland, Ml
M. Wenk. Hexabromocyclododecane Maximization Test in Guinea M96A061.1X64. Microbiological Associates, Inc. Rockville, MD. Pigs. 1996.

McDonnell, Toxicology M. 1972. Haskell Laboratory and Industrial Medicine) Report No. 18572. Haskell Laboratory for

Momma et al. Pharmacometrics, 1985,29:981-986
Nakamura et al. Contact Dermatitis, 1994, 31:72-85 Chengelis C. 1996. A 28-day repeated dose oral toxicity study WIL-186004. WIL Research Laboratories, Inc. Ashland, OH. of HBCD in rats. Study No.
Chengelis C. An Oral (Gavage) 90 Day Toxicity Study of HBCD in Rats. Study No. WIL186012. WIL Research Laboratories, Inc., Ashland, Ohio. 2001.

86J88

## 9. References Id 25637-99-4 Date 31.03.2005

(34) Zeller H and Kirsch P (1969) Hexabromocyclododecane: 28-day feeding trials with rats. BASF Unpublished Laboratory Report. (35) Zeller H and Kirsch P (1970) Hexabromocyclododecane: 90-day feeding trials with rats. BASF Unpublished Laboratory Report. (36) Gudi, R. and Schadly, E. 1996. Laboratory Study Number G96A061.342. Microbiological Associates, Inc., Rockville, MD. (37) Helleday et al. Brominated flame retardants induce intragenic recombination in mammalian cells. Mutation Research 439 (1999) 137-l 47. (38) Engelhardt, G and Hoffmann, H. (2000) Laboratory Project Identification: 26M0100/004018. Experimental Toxicology and Ecology, BASF Aktiengesellschaft, Ludwigshafen, Germany. (39) lnoue Y.K.T., Kawashima K., Uchida Y, and Monma, J. Carcinogenesis test of flame retarder Hexabromocylododecane in mice. Department of Toxicology, National Public Health Research Institute, Biological Safety Test and Research Center, Japan. (40) Stump, D. 1999. A Prenatal Developmental Toxicity Study of Hexabromocyclododecane (HBCD) in Rats. Laboratory Study No.: WIL-186009. WIL Research Laboratories, Inc., Ashland, OH. (41) Murai et al. 1985. Pharmacometrics (Japan) 29(6):981-986. (42) Chengelis C. An Oral (Gavage) 90 Day Toxicity Study of HBCD in Rats. Study No. WIL186012. WIL Research Laboratories, Inc., Ashland, Ohio. 2001 (43) Chengelis C. 1996 A 28-day repeated dose oral toxicity study of HBCD in rats. Study No. WIL-186004. WIL Research Laboratories, Inc. Ashland, OH).

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## 10. Summary and Evaluation Id Date 25637-99-4 31.03.2005

10.1 END, POINT SUMMARY

10.2 HAZARD SUMMARY

14.12.2004

10.3 RISK ASSESSMENT

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