Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study according to OECD Guideline No. 203

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Anilosäure

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Appr. 100 mL samples were taken from the middle of the test vessel at 0, 48 and 96 hours respectively.

Test solutions

Details on test solutions:
PREPARATION OF TEST SOLUTION
0.095 mL Tween 80 /L was used in the final test concneztration. After homogenizitation by ultrasound and intensive stirring the mixture was incorporated into the test vessel and stirred for another 2 hours. Concentration of the test item in the treatment vessel was determined after 0, 48 and 96 h respectively.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebrafish
- Strain: Brachydanio rerio (Hamilton-Buchanan)
- Source: West Aquarium, D-37431 Bad Lauterberg, Germany
- Age at study initiation: 3 months
- Length at study initiation (mean, range and SD): 3.0 cm, 2,8 - 3.3 cm, SD 0.16
- Feeding during test: No feeding during the test

ACCLIMATION
- Acclimation period: at least 14 days
- Type and amount of food: Tetra Min, ad libitum
- Feeding frequency: 2 times per day

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1°C
- Photoperiod (hrs dark / hrs light): 12/12
- The dissolved oxygen concentration was more than 80 % of the air saturation value


Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
Observations were carried out during exposure at the beginning of the experiment and after 2 - 4, 24, 48, 72 and 96 hours.

Test conditions

Test temperature:
21.3°C - 21.9°C (treatment), 21.3°C - 21.9°C (control)
pH:
7.1 - 8.2 (treatment), 7.7 - 8.3 (control)
Dissolved oxygen:
5.7 - 8,8 mg/L (treatment); 7.5 - 9.4 mg/L (control)
Nominal and measured concentrations:
Nominal concentration: 500 mg/L
Measured concentrations: 481.8 mg/L - 486.7 mg/L - 477.9 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass-aquaria 30 cm x 22 cm x 24 cm, 10 L per vessel
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2 (1 with solutizer)
- Biomass loading rate: fish density in the tanks was less than 1 g per litre test solution.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water, quality according ISO 7346/1. The water was saturated with oxygen. pH-value: 7.9 - 8.0.
pH , temperature and oxygen saturation were observed at the beginning, after 2-4, 24, 48, 72 and 96 h in all vessels


OTHER TEST CONDITIONS
- Photoperiod (hrs dark / hrs light): 12/12
- Light intensity: 700 Lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and behavior of the test fish and control fish were observed at 2-4, 24, 48, 72, 96 h.


Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
> 500 mg/L
Nominal / measured:
nominal
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 500 mg/L
Nominal / measured:
nominal
Basis for effect:
mortality
Details on results:
- There was no mortality in control as well as in treated fish at any observation time point.
- Behavioural abnormalities: Neither in treated nor in control fish could any abnormalities be observed at any observation time points.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this study Anilosäure did not cause any effects to zebrafish after 96 h when tested with a 500 mg/L (nominal) solution using Tween 80 as solutizer.
Executive summary:

The acute toxicity of the test item to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1984) from 1989 -10 -09 to 1989 -10 -13 at Hoechst AG, Frankfurt, Germany. A static limit test with a 500 mg/L solution, using a solubilizer was conducted. The preparation method has been chosen with regard to the test item properties and in accordance with the relevant OECD guidance document. Duration of the test was 96 h. 10 test organisms were exposed to the test solution and the control. Water quality parameters pH-value, temperature and oxygen-saturation measured after 0, 2 -4, 24, 48, 72 and 96 h were determined to be within the acceptable limits. Test item analysis was carried out at 0, 48 and 96 h. Concentrations were above 80 % of the initial concentration.

The 500 mg/L solution of the test item didn´t show any signs of toxicity nor lethality at all observation time points up to 96 hours.