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 

in the commercial product is the gamma 

performed in support 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 of the 

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 in these tests was a composite composed of 

equal parts of the members’ commercial HBCD product. BFRIP members 

are Albemarle Corporation, Dead Sea Bromine Group and Great Lakes 

Chemical Corporation. Each test article was characterized in accordance 

with Good Laboratory Practices gLidelines. The composition of the test 

article used in the 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 a test 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 of Albemarle 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 a flame retardant. Its primary application is in 

extruded (XPS) and expanded (EPS) polystyrene foam that is used as 

thermal insulation in the building industry. HBCD is highly efficient in this 

application so that very low levels are required to reach the desired flame 

retardancy. Typical HBCD levels in EPS are 0.67% and in XPS 2.5%. At 

present, HBCD is the only suitable flame retardant for these applications. 

Any other flame retardant would likely need higher load levels in the 

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 in a 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 1 for 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 in video 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 all documents 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 of the 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 in textile applications (lesser application), HBCD is ground to a fine 

particle size in order to ease its dispersion in coatings. 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) in well 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 at that 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, in order to clarify 

results reported in Walsh 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 all 3 isomers. The WAF preparations 

had serious problems with HBCD particulates that could not be resolved in 

a consistent manner by filtering or centrifugation. Thus, it was 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 

in the 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 : A technical 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 in Aerobic 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 in aerobic 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 a sandy 

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 in an 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 in the dark at 20 f 1 “C for 119 days. The concentration of 

HBCD in the 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 in the 

viable aerobic soil microcosms, compared to a 3% decrease in the abiotic 

controls, indicating that biological processes were responsible for most of 

the losses observed. Under anaerobic conditions, HBCD concentrations 

decreased 92% over 21 days in the viable microcosms compared to a less 

than 1% decrease in the 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 in the aerobic and anaerobic soils, respectively. Brominated 

degradation products were not detected in the soil or in the 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 in aerobic 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 in the 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 in the 

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 in the 

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 in the environment. The halflife of 1 -bromohexane in water at 25 “C was reduced from 20 days to 

approximately one day in 5 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 in the presence of 5 

mM HS-. Sulfide and polysulfides would be expected to be present in soils 

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 in the 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 in the formation of HS-CH2-CH2-SH, while an elimination 

reaction under alkaline conditions results in the formation of H2C=CHBr. A 

combination of elimination and substitution reactions can result in the 

formation of a mixture of HO-H2-CH2-OH and H2C=CHBr. Similar 

mechanisms may be responsible for the loss of HBCD observed in this 

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 in the 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 g soil 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 in Aerobic and Anaerobic Soil Microcosms (BFRIP and 

Dow Chemical Company) 

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

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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 in aerobic 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 a sandy 

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 in an 

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 

steam sterilization prior to the addition of HBCD. Microcosms were 

incubated in the dark at 20 f 1 “C for 119 days. The concentration of 

HBCD in the 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 in the 

viable aerobic soil microcosms, compared to a 3% decrease in the abiotic 

controls, indicating that biological processes were responsible for most of 

the losses observed. Under anaerobic conditions, HBCD concentrations 

decreased 92% over 21 days in the viable microcosms compared to a less 

than 1% decrease in the 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 in the aerobic and anaerobic soils, respectively. Brominated 

degradation products were not detected in the soil or in the 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 in aerobic 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 in the 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 in the 

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) 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-651A 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 in the 

order of F c Cl c 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 in the environment. The halflife of 1 -bromohexane in water at 25 “C was reduced from 20 days to 

approximately one day in 5 mM HS - or 0.07 

mM polysulfide (Sx 2- ). Similarly, the half-life of 1 ,Pdibromoethane in 

water at 25 “C was reduced from 1,000 days to 4 days in the presence of 5 

mM HS-. Sulfide and polysulfides would be expected to be present in soils 

at low redox potentials. Thus, reaction of HBCD with these sulfur-based 

nucleophiles may partly explain the loss of HBCD in the 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 in the formation of HS-CHBCHFSH, while an elimination 

reaction under alkaline conditions results in the formation of H2C=CHBr. A 

combination of elimination and substitution reactions can result in the 

formation of a mixture of HO-H2-CH2-OH and H2C=CHBr. Similar 

mechanisms may be responsible for the loss of HBCD observed in this 

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 ,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 in the 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) 

06.12.2004 

16188 
06.12.2004 

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 of three 

stereoisomers that are designated alpha, beta and gamma in deference 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 

of the commercial product used as test article. A rat 90-day subchronic 

study on the commercial product showed that the alpha isomer 

predominated in adipose 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 a whole 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 in the influent typically had 

no detectable HBCD. HBCD appears to settle in sludge; the gamma 

isomer typically represented some 50% of the total. Sludge from the UK 

and Ireland typically contained all three isomers, but only one STP in the 

Netherlands had detectable levels of the beta isomer in sludge. For the 5 

UK plants with measured influent, 1 had no detectable HBCD in the 

influent, 3 had gamma but no alpha or beta in the influent, and 1 had all 3 

isomers with beta predominating. In the Netherlands, for the 5 plants 

measured, 3 had alpha in the 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% of the 

total in sludge; e.g. the gamma predominated. The predominant isomer in 

the influent appears to vary with country (gamma in UK, alpha in the 

Netherlands)/ however, the dataset is extremely small and variable so this 

should be viewed with caution. 

17/88 
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 in sediment 

was gamma. 

Landfill Leachate 

Samples from the UK and Ireland had no detectable HBCD. Eleven 

samples of landfill leachate originating in the 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 in only one of the samples 

from the Netherlands. 

Whole Organisms 

Both alpha and gamma were detected in all Lake Ontario trout analyzed 

(5/5). Beta was not detected in any. Composites of 3 different 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 in the highest quantities in all 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 in freshwater 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 

in the highest quantity. Only one sample out of 11 had detectable amounts 

of all 3 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 in few 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 in waste 

water from 4 of 5 textile plants. In 1 of these 5 plants, alpha and gamma 

were present in approximately equal quantities. The gamma isomer 

18188 
3. Environmental Fate and Pathways Id 25637-99-4 

Date 31.03.2005 

predominated in soil surrounding the 5 plants. 

SUMMARY 

The published literature and unpublished data were reviewed for 

environmental monitoring results pertaining to HBCD. Only those studies 

that included specific analysis for the alpha, beta and gamma HBCD 

stereoisomers were included. Environmental matrixes reporting results for 

all three stereoisomers included sewage treatment plants, sediment, whole 

organisms, specific tissues, and textile plant wastewater, and textile plant 

soil. The gamma isomer predominated in sewage treatment plant sludge, 

sediment, landfill leachate, textile plant wastewater and textile plant soil. 

The alpha isomer was the predominant isomer detected on the analysis of 

whole organisms (lake trout, feeder fish, 3 freshwater invertebrates). 

Analysis of individual tissues produced varying results. Of the individual 

tissues analyzed, avian egg and marine mammal blubber appeared to 

produce the most consistent results. The alpha isomer was the most 

frequently detected isomer in both. Continued monitoring of egg and 

blubber may be of value, but other tissue types are not recommended. The 

most consistent data is apparently generated on the basis of whole 

organism analysis. Sewage treatment plants are effective in removing 

HBCD from the influent - effluents from plants receiving HBCD typically 

contained no detectable HBCD. 

TABLE: HBCD ISOMER SPECIFIC ANALYSIS 

A. In Various Tissues (Data from draft EU RA 2003, Gerecke 2003 and de 

Boer’s bird study 2004) 

Liver (All ppb ww) 

alpha beta gamma 

Cormarant (@kg ww) 

21 3 2 % Lipid not stated 

6 1 2 

33 1 1 

7 0.3 0 

2 0 0 

H. Porpoise (rig/g ww) 

2400 0 4 36% lipid, n=l 

H. Seal 

0 b”“‘” 0 2% lipid, n=2 or 3 

Muscle OR Fillet (All ppb ww) 

Fish 

Whiting, UK (@kg ww) 

506 247 283 % Lipid not staed 

81 61 149 

0 0 0 

Whiting, Netherlands (rig/g ww) 

0 0 0 0.6% Lipid, n=3 

Whitefish, Switzerland, (rig/g Iw) 

210 0 0 2.6%., pool of 10 

100 0 0 3.8%, pool of10 

66 0 0 1.5%, pool of 10 

54 0 0 7.2%, pool of 10 

Avian 

Sparrow Hawk (ug/kg ww) 

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 

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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 al 2004 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 of whole 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 in draft 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 dissolved phase 

0 0 

Effluent particulate phase 

0 0 

Latchingdon, UK 

lnfluent dissolved phase 

0 0 9.1 

lnfluent particulate phase 

0 0 

Sludge 205 

Effluent dissolved phase 

0 0 

Effluent particulate phase 

0 0 

Wickford, UK 

lnfluent dissolved phase 

0 0 

lnfluent particulate phase 

0 0 

Sludge 89.6 

Effluent dissolved phase 

0 0 

Effluent particulate phase 

0 0 

S. Woodham Ferrers, UK 

lnfluent dissolved phase 

0 0 0 

lnfluent particulate phase 

0 0 

Sludge 233 

Effluent dissolved phase 

0 0 

Effluent particulate phase 

0 0 

Chelmsford, UK 

lnfluent dissolved phase 

0 0 

lnfluent particulate phase 

0 0 

Sludge 541 

Effluent dissolved phase 

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 in draft 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 in draft 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 

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

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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-9 o-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 in the 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 in air, 

water, soil and sediment was estimated as 0.304, 3.26, 63.3 and 22.9%, 

respectively. The percent advected in air, 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) in air, 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%). It was expected to be 97.1% reacted in soil 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 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 

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 of the 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 in an 

EC50 of 5.2 mg/L and was within the 5 to 30 mg/L range considered 

acceptable for the test. An average of 29.1 percent inhibition was observed 

in the treatment group. 

Schaefer E and 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 301 D, 

OPPTS 3200, and Good Laboratory Practices. 

HBCD was tested for ready biodegradation in a 28 day closed bottle test at 

a concentration of 7.7 mg/L by measuring dissolved oxygen uptake and 

expressing it as a percentage of the theoretical oxygen demand or 

chemical oxygen demand. No biodegradation was observed; the percent 

biodegradation was 0. 

Schaefer. E and 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 in 2 river sediments = 11 and 32 days in aerobic 

microcosms. 

: not measured . 

. other: OECD 308 . 

. 2003 . 

. yes . 

. other TS 

: Transformation in Aerobic 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 

in aerobic 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 in Aquatic Sediment 

Systems.” Laboratory batch microcosms were prepared with authentic 

water and sediment collected from two rivers in the 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 

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 in the dark at 20 f 1 “C for 119 days. The 

concentration of HBCD in the microcosms was determined at selected time 

intervals in the 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 in the viable aerobic microcosms from both river 

systems decreased at least 90% within 21 days, while the corresponding 

decreases in the abiotic controls ranged from 7 to 62%. Disappearance of 

HBCD was observed in both the viable and abiotic anaerobic microcosms 

with the rate of loss more rapid in the viable microcosms, with HBCD 

reaching non-detected levels within 7 days. In contrast, HBCD 

concentrations in the 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 in the two river 

systems were determined to be 11 and 32 days in the aerobic microcosms 

and 1 .l and 1.5 days in the anaerobic microcosms. Brominated 

degradation products were not detected in the sediment and water layers 

or in the 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 in aerobic and 

anaerobic water/sediment microcosms. Sediment 

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 sediments although the rates were appreciable faster in the 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 in any of the 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 in these initiatives includes half-lives in soil and sediments 

ranging from 120 to 180 days. In this investigation the resulting 

biotransformation half-lives for HBCD in the two river systems were 

determined to be 11 and 32 days in the aerobic sediments and 1 .l and 1.5 

days in the anaerobic sediments, respectively. 

Limited information is available for the reactions of HBCD in the 

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 of the carbon-halogen bond, and increases in the 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 in the environment. The half-life of 1 -bromohexane in water at 25 

“C was reduced from 20 days to approximately one day in 5 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 4 days in the presence of 5 

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 in the 

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 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 ,Bdibromoethane, and 1,2-dichloroethane are 0.6, 55, and 1.8 x 104 

hours, respectively. The rapid disappearance of HBCD in the anaerobic 

sediment microcosms may be partly explained by reductive dehalogenation 

reactions. In addition, the disappearance of HBCD in the 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 in the static microcosms used in this 

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 in the 

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. 

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 in 2 river sediments = 1 .l and 1.5 days in the anaerobic 

microcosms. 

not measured 

other: OECD 308 

2003 

yes 

other TS 

: Transformation in Aerobic 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 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 

in aerobic and 

anaerobic water/sediment microcosms based on the Organization for 

Economic Co-Operation and Development (OECD) Test Guideline 308 

“Aerobic and Anaerobic Transformation in Aquatic Sediment Systems.” 

Laboratory batch microcosms were prepared with authentic water and 

sediment collected from two rivers in the 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 in an anoxic 

atmosphere (70% N2,28% C02, and 2% H2). The microcosms were preincubated 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 in the dark at 20 f 1 “C for 119 days. The 

concentration of HBCD in the microcosms was determined at selected time 

intervals in the 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 in the viable aerobic microcosms from both river 

systems decreased at least 90% within 21 days, while the corresponding 

decreases in the abiotic controls ranged from 7 to 62%. Disappearance of 

HBCD was observed in both the viable and abiotic anaerobic microcosms 

with the rate of loss more rapid in the viable microcosms, with HBCD 

reaching non-detected levels within 7 days. In contrast, HBCD 

concentrations in the 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 in the two river 

systems were determined to be 11 and 32 days in the aerobic microcosms 

and 1 .l and 1.5 days in the anaerobic microcosms. Brominated 

degradation products were not detected in the sediment and water layers 

29188 
3. Environmental Fate and Pathways Id 25637-99-4 

Date 31.03.2005 

or in the 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 in aerobic and 

anaerobic water/sediment microcosms. Sediment 

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 sediments although the rates were appreciable faster in the 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 in any of the sediment 

microcosms. 

Over the last several years a numberof 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 in these initiatives includes half-lives in soil and sediments 

ranging from 120 to 180 days. In this investigation the resulting 

biotransformation half-lives for HBCD in the two river systems were 

determined to be 11 and 32 days in the aerobic sediments and 1 .l and 1.5 

days in the anaerobic sediments, respectively. 

Limited information is available for the reactions of HBCD in the 

environment. However, the aerobic degradation and mineralization of a 

similar type of cyclic aliphatic halogenated fire retardant, FR-651 A (mixture 

of pentabromo-chlorocyclohexane, tetrabromo-dichlorocyclohexane, and 

tribromotrichlorocyclohexane) has been observed. A soil half-life of -11 

days, based upon disappearance of 14C-FR-651 A 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 of the carbon-halogen bond, and increases in the order of F < Cl c 

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 in the environment. The half-life of 1-bromohexane in water at 25 

“C was reduced from 20 days to approximately one day in 5 mM HS- or 

0.07 mM polysulfide (Sx2-). Similarly, the half-life of 1 ,Bdibromoethane in 

water at 25°C was reduced from 1,000 days to 4 days in the presence of 5 

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 in the 

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 of the carbon-radical species 

resulting from the initial electron transfer that occurs. Vicinal 

dehalogenation reactions are particularly important, and are dependent on 

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 in the anaerobic 

sediment microcosms may be partly explained by reductive dehalogenation 

reactions. In addition, the disappearance of HBCD in the 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 in the static microcosms used in this 

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 in the 

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 in Aerobic 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 in aerobic 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 a sandy 

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 in an 

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 in the dark at 20 f 1 “C for 119 days. The concentration of 

HBCD in the 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 in the 

viable aerobic soil microcosms, compared to a 3% decrease in the abiotic 

controls, indicating that biological processes were responsible for most of 

the losses observed. Under anaerobic conditions, HBCD concentrations 

decreased 92% over 21 days in the viable microcosms compared to a less 

than 1% decrease in the 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 in the aerobic and anaerobic soils, respectively. Brominated 

degradation products were not detected in the soil or in the 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 in aerobic 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 in the 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 in these initiatives includes half-lives in soil and sediments 

ranging from 120 to 180 days. In this investigation the resulting 

biotransformation half-lives for HBCD were determined to be 63 and 6.9 

days in the aerobic and anaerobic soils, respectively. Based upon these 

results HBCD is not persistent. The half-lives in soil are below the criteria 

specified by the various international protocols. 

Limited information is available for the reactions of HBCD in the 

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 

tribromotrichlorocyclohexane) was observed. A soil half-life of - 11 days 

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 be susceptible to both hydrolytic and nuclelophilic 

attack. The reactivity of halogenated aliphatic compounds increases in the 

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 

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 in the environment. The halflife of 1 -bromohexane in water at 25 “C was reduced from 20 days to 

approximately one day in 5 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 in the presence of 5 

mM HS-. Sulfide and polysulfides would be expected to be present in soils 

at low redox potentials. Thus, reaction of HBCD with these sulfur-based 

nucleophiles may partly explain the loss of HBCD in the 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 in the formation of HS-CHBCH2SH, while an elimination 

under alkaline conditions results in the formation of H2C=CHBr. A 

combination of elimination and substitution reactions can result in the 

formation of a mixture of HO-H2-CH2-OH and H2C=CHBr. Similar 

mechanisms 

study. 

may be responsible for the loss of HBCD observed in this 

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 in the 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 in soil are 

in various 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 in Aerobic and Anaerobic Soil Microcosms (Sponosr: ACC 

33188 
3. Environmental Fate and Pathways Id 25637-99-4 

Date 31.03.2005 

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 in aerobic 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 a sandy 

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 preincubated 

at 20 f 1 “C for 35 days. Anaerobic microcosms were prepared in an 

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 

steam sterilization prior to the addition of HBCD. Microcosms were 

incubated in the dark at 20 f 1 “C for 119 days. The concentration of 

HBCD in the 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 in the 

viable aerobic soil microcosms, compared to a 3% decrease in the abiotic 

controls, indicating that biological processes were responsible for most of 

the losses observed. Under anaerobic conditions, HBCD concentrations 

decreased 92% over 21 days in the viable microcosms compared to a less 

than 1% decrease in the 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 in the aerobic and anaerobic soils, respectively. Brominated 

degradation products were not detected in the soil or in the 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 in aerobic 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 in the 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. 

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 in these initiatives includes half-lives in soil and sediments . was 
3. Environmental Fate and Pathways Id 25637-99-4 

Date 31.03.2005 

ranging from 120 to 180 days. In this investigation the resulting 

biotransformation half-lives for HBCD were determined to be 63 and 6.9 

days in the aerobic and anaerobic soils, respectively. Based upon these 

results HBCD is not persistent. The half-lives in soil are below the criteria 

specified by the various international protocols. 

Limited information is available for the reactions of HBCD in the 

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 in the 

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 in the environment. The halflife of 1 -bromohexane in water at 25 “C was reduced from 20 days to 

approximately one day in 5 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 in the presence of 5 

mM HS-. Sulfide and polysulfides would be expected to be present in soils 

at low redox potentials. Thus, reaction of HBCD with these sulfur-based 

nucleophiles may partly explain the loss of HBCD in the 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 in the formation of HS-CH2-CH2-SH, while an elimination 

reaction under alkaline conditions results in the formation of H2GCHBr. A 

combination of elimination and substitution reactions can result in the 

formation of a mixture of HO-H2-CH2-OH and H2C=CHBr. Similar 

mechanisms may be responsible for the loss of HBCD observed in this 

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 in the anaerobic soil 

microcosms may be partly explained by reductive dehalogenation 

reactions. 

Based upon these results, HBCD is not persistent. Its half-lives in soil are 

clearly below the criteria for persistence specified in various 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 in anaerobic 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 of the 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 in activated 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 3 to 5 mg/kg (or mg/L), exceeding 

the water solubility of the 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 3 stereoisomers 

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 in the 

anaerobic digester sludge and in freshwater aerobic and anaerobic 

sediment microcosms. Conversely, no degradation of HBCD was noted in 

the soil microcosms incubated under aerobic conditions. In the digester 

sludge and sediment, degradation of each of the three stereoisomers 

occurred over the course of the study. Little difference was noted in the 

disappearance of the three stereoisomers, indicating similarity in the 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 in the sludge and sediment test 

mixtures was formation of three [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 

in wastewater 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 in sludge, 

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 of the 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) in whole 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 in tissue 

concentrations at day 35. The unexpected increase in tissue 

concentrations on day 35 may have been due to the variability in the 

measured water concentrations in this treatment group. The variability in 

turn is likely a function of the extremely low nominal concentration at this 

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 in the nominal 3.4 

ug HBCD/L treatment group is considered a better estimate than that in the 

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 in this 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 in the 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 in the 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) 

38188 
27.12.2004 

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 11 and 

32 days in the 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 in soil 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 limit of 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 in the 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 all 3 

HBCD diastereomers; however, the trace residues of the alpha and beta 

diastereomers were evident in the 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 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 of toxicity, 

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): a 48- 

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 EC1 0, 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): A 96Hour 

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 of water-soluble 

components of hexabromocyclododecane (HBCD) to the marine diatom 

Skeletonema costatum, using a saltwater 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 of the 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 in salwater 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 a solid support coate with HBCD to achieve the test 

concentraiton of HBCD in saltwater algal medium. Measured test 

concentrations were determined from samples of test medium collected 

from teh treament and each control group at the 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 of the cell densities, biomass and growth rates, as well as 

examination of the concentration-response pattern. 

The analytical method for the 3 HBCD stereoisomers in saltwater algal 

medium was based on methodology developed by Wildlife International 

LTD. It used an HPLC interfaced with a triple quadrupole LC mass 

spectromer operated in single-quadrupole selective ion monitoring mode. 

The LOCI was 0.5 ug a.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 .O ug a.i./L, respectively. Arithmetic mean measured test 

concentrations were used to calculate the NOEC values. 

S. costatum were exposed to a single concentraion of 41 .O ug 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 in the treated group was 19, 21 and 

7.3%, respectively, relative to control no column. Similarly, relative to the 

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 .O I.@ a.i./L and > 41 .O ug 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): a 72-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 3 species of marine 

algae in 6 different 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 I 88 
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 of the 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 

of the test was further supported by the results from the 3,5-dichlorophenol 

reference group, which resulted in an EC50 of 5.2 mg/L and was within the 

5 to 30 mg/L range considered acceptable for the test. An average of 29.1 

percent inhibition was observed in the 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 in the 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 all 3 

HBCD diastereomers; however, the trace residues of the alpha and beta 

diastereomers evident in the 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 

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 of toxicity, 

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 in this 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). All three species were tested at 50, 100, 500, 1000 mg/Kg dry 

sediment in two different types sediments, one with a 2% organic carbon 

content and the other with a 5% organic carbon content. Based on the 

results of the range-finding studies, the amphipods were found to be the 

most sensitive species in both sediment types, with clear effects in the 500 

mg/Kg treatment group. This reports the results of HBCD exposure in the 

5% organic content sediment in Hyalella azteca. A similar study in 2% 

organic carbon was also performed. 

Groups of amphipods were exposed to a geometric series of six test 

concentrations and a negative control for 28 days under flow-through test 

conditions. Eight replicate test compartments were maintained in each 

treatment and control group, with 10 amphipods in each test compartment, 

for a total of 80 amphipods per test concentration. Each test compartment 

contained a quantity of sediment and overlying water. Additional replicates 

were added in the 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 of the 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 in the sediment, pore water and overlying 

water samples collected during test confirmed that the test article tended to 

remain in sediment and not move into the pore water or overlying water. 

Concentrations in sediment at the highest dose level ranged from 78.2 -

122% of nominal, while the lowest dose level ranged from below the limit of 

quantitation (12.5 mg/kg) to 197% of nominal. All overlying 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 in the 

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 in the 0,31,63, 125,250 and 500 mg/kg groups. Fungal 

growth was noted in all replicates. 

The mean number of amphipods in the negative control, 31, 63, 125, 250, 

500 and 1000 mg/Kg treatment groups at test termination was 9.1, 8.6, 5.9, 

6.1, 7.0, 8.5 and 9.1, respectively. The mean numbers of amphipods in the 

63, 125 and 250 mg/Kg treatment groups were found to be statistically 

different (p < 0.05) from the negative control group. Survival in the 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 in the number of organisms present at test termination in 

comparison to the negative control group was less than 50% in all 

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 in the 63, 125, and 250 mg/Kg treatment groups 

was 35.2, 33.0, and 23.1%, respectively. The mortality in these groups was 

moderate and there were clearly no effects at the 500 and 1000 mg/Kg 

treatment levels. Therefore, the mortality observed in the middle test 

concentrations was not considered treatment related since there was no 

evidence of a concentration dependent response. 

The average dry weight per amphipod in the negative control group was 

0.19 mg. The average dry weight per amphipod in the 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 no apparent effects on 

growth (dry weight) observed at test termination. 

The 28-day EC50 value for amphipods (Hyalella azteca) exposed to 

hexabromocyclododecane in sediment 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 in this 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) 

: Prolonged Sediment Study with Hyalella azteca, 2% 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 

EPA, 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). All three species were tested at 50, 100, 500, 1000 mg/Kg dry 

sediment in two different types sediments, one with a 2% organic carbon 

content and the other with a 5% organic carbon content. Based on the 

results of the range-finding studies, the amphipods were found tb be the 

most sensitive species in both sediment types, with clear effects in the 500 

mg/Kg treatment group. This reports the results of HBCD exposure in the 

5% organic content sediment in Hyalella azteca. A similar study in 5% 

organic carbon was also been performed. 

Groups of amphipods were exposed to a geometric series of six test 

concentrations and a negative control for 28 days under flow-through test 

conditions. Eight replicate test compartments were maintained in each 

treatment and control group, with 10 amphipods in each test compartment, 

for a total of 80 amphipods per test concentration. Each test compartment 

contained a quantity of sediment and overlying water. Additional replicates 

were added in the 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 of the 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 

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 in the sediment, pore water and overlying 

water samples collected during test confirmed that the test article tended to 

remain in sediment and not move into the pore water or overlying water. 

Concentrations in sediment at the highest dose level ranged from 82.8 - 115% of nominal, while the lowest dose level ranged from 49.5 to 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 limit of 

quantitation, but those from the highest dose level, 1000 mg/kg sediment, 

had HBCD concentrations in the 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 in the 0,31 and 1000 mg/kg groups. Fungal growth was 

noted in all replicates. 

The mean number of amphipods in the 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 in the 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 in a 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 a statistically 

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 all seedlings in this 

study was determined to be 5,000 mg of HBCD/kg, which was the highest 

nominal soil concentration tested. 

Porch et al. 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 in this 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. 

After 28 days of exposure to HBCD, percent mortality of the adult worms 

was 0, 5, 0, 0, 0, 0, 3, and 0% in the 0, 61.2, 145, 244, 578, 1,150, 2,180, 

and 4,190 mg HBCD/kg groups, respectively. All of the live earthworms 

were normal in appearance 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 in the 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 3 235 mg HBCD/kg. 

Using the estimated soil Koc and the formula provided in the 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 in the 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 of the 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 in this 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 of the adult worms 

was 0, 5,0, 0, 0, 0, 3, and 0% in the 0, 61.2, 145,244, 578, 1,150, 2,180, 

and 4,190 mg HBCD/kg groups, respectively. All of the live earthworms 

were normal in appearance 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 in the 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 3 235 mg HBCD/kg. 

Using the estimated soil Koc in Table 1 and the formula provided in the 

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 in the 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 : in vivo 

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, in a report by Dean and Leong 

(1977), rats exposed dermally to a high dose of HBCD in saline 

experienced diarrhea and slight weight loss. This finding indicates that at 

least some absorption occurs via the dermai route. 

In an unpublished study by Vesicol Chemical Corporation (1980), rats 

administered a single 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 in the feces and urine following absorption, 

with 70% of the administed radioactivity eliminatd in the feces and another 

16% eliminated in the urine 72 hr after dosing. A two-compartment model 

was constructed, with non-adipose tissues in one compartment and 

adipose tissue in the 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 in the 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) in olive oil at 500 mg/kg-d for 5 d. HBCD was found 

to be present only in adipose tissue, and in none of the other organs 

examined (i.e., spleen, pancrease, liver, kidneys, and heart). HBCD was 

found to be excreted in the feces, with an average of 32-35% of the 

cumulative administered dose excreted. No HBCD was found in the urine. 

Although differences in study design, including the test vehicle and the 

analytic methods used, may account for some of the difference in the 

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 in the 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 : In vivo 

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 : 5 day(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 in olive oil was prepared by miing well to 

homogeneity in a 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 in the fecal 

and organ homogenates. 

The average daily rate of excretion in the 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 in the intestinal tissue and the 

amount remaining in the lumen loop “small”. 

The test article was detected only in the adipose tissue after dosing for 5 

days. The level in adipose 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 : A single 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 in the 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 4 animlas 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 in feces (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 in feces 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 in adipose 

tissue. 

In females at 8, 24, 48 and 72 hours post-dosing, the total 14Cactivity 

detected in tissues 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% of the dose. Similarly, 

that detected in feces from female rats was -4, 65, 54 and 77% at 8,24, 48 

and 72 h. Urine contained 0.1, 6, 18 and 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 in feces and urine of female 

rats. Thus, at 48 hours post-dosing approximately 86% of the dose was 

recovered in the 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 at 24 hours, had a total fecal 14Ccontent 

of 65% of the dose, and females sacrificed at 72 hours had a total 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. It appears 

likely that some unknown factor resulted in the lower O-24 hour percent 

recovery from feces, and that percent of dose present in tissues, 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 of the 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 et al. 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 of 200 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 1 Cday 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 at 4 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 in the 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 in ear 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 .O at all doses tested. Lymphocyte proliferation by DMF, vehihcle 

treated mice (2015 dpm) was higher than historical laboratory values 

commonly observed using acetone and olive oil as a vehicle. This is not 

inconsistent with that reported in the 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 in this study. On the basis of these results, HBCD would not be 

considered to have contact sensitization potential. 

Woolhiser M and 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 in 2 stages (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% in the 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 a two 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 in any 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 

a two-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 of the 

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 in guinea 

pigs, one in mice, and one in human volunteers. The 1997 guinea pig 

maximization test performed by BFRIP was negative. Two studies in the 

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 in the 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 in these 3 sensitization tests are 

provided in the table below. 

Comparison of the methodology used in 3 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 mg 

0.00025,0.0025 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, in the 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 6 females in the 125 and 350 mg/kg/day groups and 12 males 

and 12 females in the 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 6 animals/sex in the control and 1000 mg/kg/day groups 

remained on-test untreated for a 1Cday recovery period. At the end of the 

recovery period, all animals 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 

at the 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 

all animals were weighed at each sacrifice. Approximately 40 tissues were 

collected and preserved at each necropsy from all animals. 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 in all dose levels. 

Survival was not affected by administration of the test article. All animals 

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 I 88 
5. Toxicity Id 25637-99-4 

Date 31.03.2005 

Body weights, weight gain and food consumption of treated animals were 

compared statistically by sex and treatment day to their respective control 

groups (p < 0.05 or 0.01) and were not affected by treatment. No 

statistically significant differences in body weight between control and 

treated animals were detected with the exception of an increase in mean 

female body weight in the 350 mg/kg/day group during week 2 of 

treatment. Mean female body weight at that time point was 196 g versus 

179 in the control group. No statistically significant differences in body 

weight gain between control and treated animals were detected with the 

exception of a decrease in mean male body weight gain in the 1,000 

mg/kg/day recovery group during week 1 of recovery. Mean male body 

weight gain at that time point was 21 g versus 31 in the control group; 

mean male body weight was not statistically different from the control 

mean. No statistically significant differences in food consumption between 

control and treated animals were detected with the exception of an 

increase in mean female food consumption in the 350 mg/kg/day group 

during weeks -1, 1, and 2 of treatment. Mean female food consumption at 

that those time points were 18, 17 and 17 g versusl6, 15 and 15 g in the 

control group, respectively. 

Functional observation battery and motor activity results from treated 

animals were compared statistically by sex and treatment day to their 

respective control groups (p c 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. 

No statistically significant differences between treated and control animals 

were found for hematology parameters with the exception of an increase in 

the mean activated partial thromboplastin time in the 1000 mg/kg/day 

males on week 4 and a decrease in the mean prothrombin time in the 1000 

mg/kg/day females on week 4. These statistical differences were not of 

toxicological significance. 

No toxicologically significant effects on serum chemistry values related to 

test article administration were observed at the 28-day primary and 42-day 

recovery sacrifice. Scattered instances of statistically significant 

differences between treated and control animals were detected for some 

serum chemistry parameters at the 28-day primary sacrifice. These 

scattered statistical differences were not considered toxicologically 

significant because the statistical differences occurred: in the absence of a 

dose response, in the absence of the accompanying clinical chemistry 

changes expected, in the opposite direction from what occurs in a toxic 

state, in a direction which is without physiologic significance, or due to 

potential interference with the laboratory method. No statistically significant 

differences in serum chemistry parameters were detected between groups 

at the 42-day recovery sacrifice. 

No gross lesions that could be attributed to the test article were detected at 

either necropsy. Gross lesions were nonspecific, low in incidence, nondose-related and considered incidental. 

No microscopic lesions that could be attributed to the test article were 

detected on histopathologic exam. Microscopic changes were nonspecific, 

low in incidence, non-dose-related and considered incidental. 

No statistical significant differences in organ weight or organ to body weight 

ratios were detected between control and treated animals with one 

exception. Absolute liver weights were statistically significantly increased 

with respect to control means at the 28-day sacrifice in males in the high 

dose and females in the mid and high dose. Liver to body weight ratios in 

mid and high dose males and low, mid and high dose females were 

statistically significantly increased at the 28-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 in absolute 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. In the absence of test article related 

histopathologic and serum chemistry changes, increases in liver weight are 

considered an adaptive, rather than a toxic 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, concurrent vehicle

: = 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 of three lots of commercial 

hexabromocyclododecane (HBCD), was administered by oral gavage in 

corn oil once 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, 10 animals/sex/group 

were euthanized and necropsied. The remaining rats continued on test 

untreated for a 28-day recovery period prior to necropsy. 

In addition to the main toxicology groups, two satellite groups of 20 

animals/sex/group were treated concurrently in an 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 in the 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 13 and 17, for urinalysis. Ocular examinations were performed prior 

to study initiation and during study weeks 12 and 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 all animals. 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 in the 100 

and 300 mg/kg/day doses, and gross lesions from all animals 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 lipid acoumulation 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 in the 

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 in albumin (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 in the 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 in serum 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 at the end of the recovery 

period. The increased serum albumin and gamma glutamyltransferase 

68186 
I 

5. Toxicity Id 25637-99-4 

Date 31.03.2005 

levels were considered related to the increased in liver weight in treated 

animals that was believed due to enzyme induction. Hepatic GGT is 

inducible in the 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 in the 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 in serum 

chloride was considered secondary to be presence of free bromide in the 

test article preparation that interfered with the chloride determination 

methodology. The decrease in T4, which was also reversible, was 

considered related to the increased liver weights in that induction of hepatic 

UDPG is known to enhance the elimination of T4. 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 

of frank 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 in females in the 

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 in the 300 (l/l 0) and 1000 mg/kg/day (2/l 0) dose groups. 

Minimal to mild hepatocellular hypertrophy was also detected only in the 

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 in the control, 

100,300 and 1000 mg/kg/day groups, respectively and in 4/l 0 and 3/l 0 

females in the 300 and 1000 mg/kg/day groups respectively. In addition, 

mild thyroid follicular hypertrophy was detected in 4/10 females in the 1000 

mg/kg/day group. 

The histologic changes in the 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 in all dose 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 in this study, an increase in liver weight, the lack of 

adverse histologic effects in the liver and the apparent normal functioning 

of the 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 in this 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 in all treated groups without delayed or long-tern toxic effects. 

Limited pharmacokinetic studies indicate HBCD is extensively metabolized 

prior to excretion in the 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 in adipose 

tissue at all time points were alpha>>gamma>beta in contrast to the test 

article composition of gamma>>alpha>beta.) Thus, it should 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 of time, as was the case in the 

The histologic changes in the thyroid consisted of a slight increase in the 

incidence of minimal follicular cell hypertrophy in the high dose males and 

minimal or mild hypertrophy in the high dose females. It was 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 is the typical 

adaptive response of a healthy normally functioning organism acting to 

maintain serum T4 levels in the normal range. 

The mean prostate weight was increased in the 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 in the adipose tissue of male and female rats treated 

with 1000 mglkglday for up to 90 days. Isomer-specific analysis showed 

that the relative isomer concentrations in adipose tissue at all time 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 all time points 

and declined during the recovery period. 

All the 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 a toxic 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, if any, 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 oil for 

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% of the diet for 28 days. Doses calculated from 

the actual body weights and food consumption in this study are 0, 940, 

2410, and 4820 mg/kg body weight/day. 

No clinical signs related to treatment were observed at the 1% dose level. 

Body weights at the 1 and 2.5% dose levels were comparable to the 

controls. Liver weights (absolute and relative to body weight) were 

increased at all dose levels, but no microscopic pathology was detected. 

Thyroid hyperplasia was observed in some animals at all doses, and “very 

slight numerical development of the follicles and ripening follicles in the 

ovaries of females” at the high dose (4820 mg/kg/d) was reported. No 

changes in any other organ related to treatment and no changes in clinical 

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 in view 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 in this 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 in clinical chemistry results. An 

increase in the relative liver to body weight ratio was found, and was 

accompanied by fatty accumulation but no other histologically discernible 

changes were detected in the liver. Further, no histological changes were 

found in any other organ. The original report stated that in the “absence of 

detectable clinico-chemical disturbances or histological changes of the vital 

organs, it was 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 in the 

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% of the 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 in the 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 of two lots of FM-100, Lot 

53 and residue of Lot 3322 in the 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) In vitro 

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 : In vitro 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 in the in vitro mammalian cytogenetic test using human 

peripheral blood lymphocytes both in the absence and presence of 

metabolic activation. The assay was performed in two phases. The first 

phase, the initial chromosome aberration assay, was conducted to 

establish the dose range for testing and to evaluate the clastogenic 

potential of the test article. The second phase, the independent repeat 

chromosome aberration assay, was performed to confirm the test system 

response to the test article seen in the initial assay. 

Dimethylsulfoxide was used as a solvent. In the initial assay, the maximum 

dose tested was 2,500 @ml. Dose levels greater than 2,500 ug/ml were 

insoluble in treatment 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 in treatment 

medium at all other doses tested. In the non-activated portion of the initial 

assay cells were exposed to the test article continuously for 20 hours; in 

the SS-activated portion of the 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 in the nonactivate study and 750 and 2,500 ug/ml in the 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, in the non-activated 

study. In the 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 in the 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 in treatment medium at 

dose levels 150 ug/ml and higher. The test article was soluble in treatment 

medium at all other concentrations tested. In the 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 in the 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 

of treatment. Toxicity, measured by mitotic inhibition, was -55% and 94% 

at the 20 and 44 hr harvests, respectively, at the highest dose levels (600 

and 300 @ml) evaluated for chromosome aberrations in the nonactivated 

studies. In the 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 in the non-activated 44 hr harvest and in the SS-activated 

20 hr harvest was not analyzed for chromosome aberrations due to an 

insufficient number of storable metaphase cells. No statistically significant 

increases in structural 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 in this study exhibit a spontaneous partial duplication of the 

hprt gene, resulting in a non-functional hgprt protein. These mutants revert 

spontaneously to a functional hprt gene phenotype by recombination with a 

frequency of 1 x 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 at 37 degrees C. 

HBCD was tested in vitro in hamster cells (Sp5N79 and SPD8) in a 

recombination assay at five doses between 2 and 20 @ml plus a control. 

In the SPD8 cells, HBCD concentrations of 0, 3, 6, 10, 15, and 20 &j/ml 

resulted in a reversion frequency of 1 .O, 0.7, 0.8, 0.9, 1.4, and 1.9, 

respectively. Cytoxicity was observed at the 20 @ml dose. In the Sp5 

cells, HBCD concentrations of 0, 2, 5, 10, 15, 20 @ml resulted in a 

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 t test, ~~0.05). Treatment with HBCD resulted in an - 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 a test 

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 the vehicle, 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 in the course of mitosis. HBCD was clearly negative for 

clastogenicity and the ability to induce spindle poison effects in this 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 by the 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, 7 days/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 in the 

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 oil only. 

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 all fetuses, and the sex and individual weights of fetuses 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 I 88 
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 a developmental . 

. other . 

. 1985 .

. no data . 

. other TS 

toxicant 

: Murai et al. 1985 (Pharmacometrics (Japan) 29(6):981-986) identified no 

reproductive or developmental effects in the rat at doses up to 1% in the 

diet administered from days O-20 of gestation. This dose is approximately 

equivalent to 500 mg/kg/d. 

The Murai et al study consisted of a 7 day dose range finding study (n=5 

rats/dose group) and a combined teratogenicity-developmental study 

(n=20/dose group). Doses in the 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 > 1 g/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 of the 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 in the 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 in the 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, live fetuses, sex ratio, or body or placental weight. 

No fetal deaths occurred in any group. No external, skeletal or visceral 

malformations were detected. A few skeletal variations were detected but 

where of similar types and numbers in the control and treated groups. 

There was no significant differences between the control and treated 

groups in the number of implantation, live newborns, dead newborns, 

newborn parturition index. The weaning and survival index was 

comparable in the control and treated groups. Body weight changes 

newborns was comparable in all groups. 

live 

in the 

No reproductive or developmental effects where detected in rats at HBCD 

doses up to 1% in the 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 6 wks. NOEL = 1% of diet. 

Reliability 

28.12.2004 

: 

Murai, T. Kawasaki, H., Kanoh, S. 1985. Studies on the toxicity of 

insecticides and food additives in pregnant 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 : In vivo 

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 in the 

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 in a second 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 all demonstrate HBCD has no effect on the reproductive system at 

the limit dose of 1,000 mglkgld. 

In the 2001 90-Day study, considered the most informative of the 

subchronic studies mentioned in the 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 in animals in the 

control and high dose groups. Vaginal smears for determination of the 

stage of estrus were obtained form all females 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 in females 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 in weight or 

microscopic effects were noted in the organs of the reproductive tract with 

the sole exception of an increase in weight of the prostate at 1,000 mg/kg/d 

on day 90. Mean prostate weight was increased in the 1,000 mglkgld dose 

group compared to the control mean after 90 days of treatment (0.95, 0.99, 

1.12, 1.25* g in the 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 in the control and high dose groups were not 

statistically different. No test article-related changes in the 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 oil once 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 in the 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, in the 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 6 females in the 125 and 350 mg/kg/day groups and 12 males 

and 12 females in the 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 6 animals/sex in the control and 1000 mg/kg/day groups 

remained on-test untreated for a 14-day recovery period. At the end of the 

recovery period, all animals 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 in the 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 

81 I88 
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 in that 

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 in various matrixes. (See the studies sponsored by BFRIP 

in 2003 and 2004.) 

Reliability : (1) valid without restriction 

29.12.2004 

6.2 DETECTION AND IDENTlFICAT’lON 

82188 
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 

83188 
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 

84 I 88 
9. References Id 25637-99-4 

Date 31.03.2005 

(1) Albemarle Corporation. Saytex HP-900 Flame Retardant. Technical Data Sheet. BC1033 

(1 l/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 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. 

(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, E and Haberlein, D., 1996, Hexabromocyclododecane (HBCD): Closed Bottle 

Test. Project No.: 439E-102. Wildlife International Ltd. Easton, MD. 

(8) 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. 

(9) Davis JW, Gonsior SJ, Markham DA, and Marty GT. 2004. Investigation of the 

biodegradation of [14C]hexabromocyclododecane in sludge, 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): A 96-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): a 48-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 of the 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). 

87188 
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 

88188