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Ecotoxicological information

Short-term toxicity to fish

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Administrative data

short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
05 May - 19 June 2009
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
according to guideline
OECD Guideline 203 (Fish, Acute Toxicity Test)
according to guideline
EU Method C.1 (Acute Toxicity for Fish)
GLP compliance:
yes (incl. QA statement)
The Department of Health of the Government of the United Kingdom

Test material

Details on test material:
- Name of test material (as cited in study report): Benzene, 1,1 ‘-(1‚2-ethanediyl)bis-, brominated, [CAS No. 1092834-40-6]; product name: SH-1; reaction product of bromine and 1,2-diphenylethane
- Substance type: white powder
- Physical state: solid
- Analytical purity: >99%
- Composition of test material, percentage of components: Penta bromodiphenylethane 0-2%, Hexa bromodiphenylethane 40-80%, Hepta bromodiphenylethane 1-20%, Octa bromodiphenylethane 1-30%, Nona bromodiphenylethane 0-10%, Deca bromodiphenylethane 0-5%
- Lot/batch No.: 20090105
- Expiration date of the lot/batch: 08 January 2011
- Storage condition of test material: room temperature in the dark

Sampling and analysis

Analytical monitoring:
Details on sampling:
The concentration and stability of the test material in the centrifuged and uncentrifuged samples were verified by chemical analysis at 0 (fresh media), 24, 48, 72 (old and fresh media) and 96 hours (old media).

Test solutions

Details on test solutions:
An amount of test material (100 mg) was dissolved in tetrahydrofuran and the volume adjusted to 25 mL to give a 100 mg/25 mL solvent stock solution. Serial dilutions were prepared from this to give a 0.40 mg/10 mL solvent stock solution. An aliquot (2.0 mL) of the 0.40 mg/25 mL solvent stock solution was dispersed in dechlorinated tap water and stirred with a flat bladed mixer for approximately 1 minute to give the required test concentration. This method of preparation was conducted in duplicate to give replicates R1 and R2.

Each of the stock solutions were inverted several times to ensure adequate mixing and homogeneity.

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
- Common name: rainbow trout
- Source: Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK
- Age at study initiation (mean and range, SD): juvenile
- Method of breeding: maintained in-house since 20 May 2009; in glass fibre tank with a"single-pass" water renewal system
- light: 18 hours light, 8 hours dark, with 20 min dawn and dusk transition periods
- Feeding during test: no; stock fish were fed commercial trout pellets which was discontinued 24 hours prior to the start of the definitive test

- Acclimation period: 03 - 15 June 2009
- Acclimation conditions (same as test or not): yes

Study design

Test type:
Water media type:
Limit test:
Total exposure duration:
96 h

Test conditions

test water: approx. 140 mg/L as CaCO3
Test temperature:
14 - 15°C
7.6 - 8.0
Dissolved oxygen:
9.8 - 10.1 mg O2/L
Nominal and measured concentrations:
0.0040 mg/L (nominal)
0.0034 mg/L (time-weighed mean measured)
Details on test conditions:
- Test vessel:
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 20 L glass vessels
- Aeration: via narrow bore glass tubes
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

- Source/preparation of dilution water: laboratory water was dechlorinated by passage through an activated carbon filter and partly softened, giving water a total hardness of approx. 140 mg/L as CaCO2. Water was then heated to achieve required temperature.
- Total organic carbon: 1.041 mg/L
- Pesticides: 0
- Chlorine: 0.297 mg/L
- Conductivity: 421.385 µS/cm at 20°C
- Intervals of water quality measurement: reporting period 01 Jan - 31 Dec 2007

- Photoperiod: 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours.

- Justification for using less concentrations than requested by guideline: see "any other information on materials and methods"
- Range finding study
- Test concentrations: single nominal concentration of 0.0040 mg/L (as the results from a Acute Toxicity to Daphnia magna test, Harlan Laboratories Ltd Project Number: 2737/0002)
- Results used to determine the conditions for the definitive study: see "any other information on materials and methods"
Reference substance (positive control):

Results and discussion

Effect concentrationsopen allclose all
96 h
Dose descriptor:
Effect conc.:
0.003 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
96 h
Dose descriptor:
Effect conc.:
> 0.003 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
no mortalities in 14 fish exposed, no sublethal effects

Any other information on results incl. tables

Sublethal observations / clinical signs:

Current regulatory advice is that in cases where a decline in measured concentrations is observed, time-weighed mean measured concentrations should be used for calculating LC50 values. It was therefore considered justifiable to base the results on the time-weighted mean measured test concentrations in order to give a "worst case" analysis of the data. 0.0034 mg/L was the highest attainable test concentration that could be prepared due to the limited solubility of the test material.

Applicant's summary and conclusion

Validity criteria fulfilled:
Based on the time-weighted mean measured test concentrations of the centrifuged test media the acute toxicity of the test material to rainbow trout gave a 96-hour LC50 value of greater than 0.0034 mg/L. The No Observed Effect Concentration was 0.0034 mg/L.