Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989
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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
84/449/EEC
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
no

Test solutions

Details on test solutions:
PREPARATION OF TEST SOLUTION
The test item was diluted with water. After homogenizitation by ultrasound and intensive stirring the mixture was incorporated into the test vessel and stirred for another 2 hours. At all concentration levels precipitate were observed.

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: ca. 3 months
- Length at study initiation (mean (n=60), range and SD): 3.0 cm, 2,5 - 3.5 cm, SD 0.27
- 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
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. Observation of symptoms was restricted by white turbidity due to low solubillity of the test substance.

Test conditions

Test temperature:
21.0°C - 21.9°C (treatment), 21.1°C - 23.0°C (control)
pH:
6.7 - 8.2 (treatment), 7.4 - 8.4 (control)
Dissolved oxygen:
6.3 - 9,3 mg/L (treatment); 6.7 - 9.6 mg/L (control)
Nominal and measured concentrations:
Nominal concentrations: 1 000, 2 500, 3 500, 5 000, 7 100 and 10 000 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): 1
- 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:
LC0
Effect conc.:
>= 1 000 mg/L
Nominal / measured:
nominal
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 10 000 mg/L
Nominal / measured:
nominal
Basis for effect:
mortality
Details on results:
- There was no mortality in control at any observation time point.
- At concentrations 2 500 - 10 000 mortalities were observed after 24 h.
- Behavioural abnormalities at all concentration levels: swimming posture tail- and head-heavy, upside-down and on side; increased and decreased activity; increased and decreased fright reaction; swimming and sitting at the survace and on the base of the tank; projecting opercula; convulsions, twitching,

Any other information on results incl. tables

Table 1: test concentrations and results

Concentration

Mortality

[mg/L]

2 - 4 h

24 h

48 h

72 h

96 h

0

0 / 10

0 / 10

0 / 10

0 / 10

0 / 10

10 000

0 / 10

2 / 10

3 / 10

3 / 10

3 / 10

0

0 / 10

0 / 10

0 / 10

0 / 10

0 / 10

7 100

0 / 10

1 / 10

2 / 10

2 / 10

2 / 10

0

0 / 10

0 / 10

0 / 10

0 / 10

0 / 10

5 000

0 / 10

2 / 10

2 / 10

2 / 10

2 / 10

0

0 / 10

0 / 10

0 / 10

0 / 10

0 / 10

3 500

0 / 10

0 / 10

1 / 10

2 / 10

2 / 10

0

0 / 10

0 / 10

0 / 10

0 / 10

0 / 10

2 500

0 / 10

0 / 10

1 / 10

1 / 10

1 / 10

0

0 / 10

0 / 10

0 / 10

0 / 10

0 / 10

1 000

0 / 10

0 / 10

0 / 10

0 / 10

0 / 10

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
67/548/EEC: Acute aquatic toxicity: no classification warranted
1272/2008/EC: Acute aquatic toxicity: no classification warranted
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) at Hoechst AG, Frankfurt, Germany. Static test with the concentrations 1 000, 2 500, 3 500, 5 000, 7 100 and 10 000 mg/L solution were 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 respectively. 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.

There were no mortalities at a concentration of 1 000 mg/L at any observation time point. At all other concentration levels up to 30 % mortalities were observed after 24 h to 72 h.

At all test concentration levels the following signs of toxicity were observed: swimming posture tail- and head-heavy, upside-down and on side; increased and decreased activity; increased and decreased fright reaction; swimming and sitting at the survace and on the base of the tank; projecting opercula; convulsions, twitching.

The median lethal concentration of the test item to zebrafish observed is:

LC50 (96 h): greater than 10 000mg/L