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Administrative data

short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed according to relevant guidelines and compliant to GLP

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
other: OCDE n° 203 (1984) Directive CEE 84/449 (1984) - annexe V - méthode C1
GLP compliance:

Test material

Sampling and analysis

Analytical monitoring:

Test solutions


Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)

Study design

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

Test conditions

90 mg CaCO3/L
Test temperature:
20.2°C - 22.6°C
6.63 - 7.47
Dissolved oxygen:
at least 82% of the air saturation value
Nominal and measured concentrations:
limit concentration: 1000 mg/L (nominal)
Details on test conditions:
The test media were heterogeneous supensions of the test substance. These suspensions were prepered approx. 72 hours prior to the start of the exposure, submitted to magnetic stirring for 1h 45 min and allowed to settle until the beginning of the exposure. The test substance particles forming deposit at the tank bottom and onto glass tank surfaces, and floating at the surface of the experimental water. Coloration of the test media was due to test substance in suspension.
Periodic manual stirring and air bubbling allowed, as far as possible, dispersion of the test substance particles in the test media throughout the study period.
Reference substance (positive control):

Results and discussion

Effect concentrations
96 h
Dose descriptor:
Effect conc.:
> 1 000 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: theoretical concentration
Details on results:
No mortality was recorded during the 96-hour study periode, until the theoretical concentration of 1000 mg/L. The water solubility of the test item is < 0.1 mg/L. Therefore, no mortality of the saturated solution was recorded during the 96-hour study periode.

Applicant's summary and conclusion

Validity criteria fulfilled:
LC50 - 96 hours for fish in freshwater medium: higer than 1000 mg/L
Executive summary:

Freshwater fish (Brachydanio rerio) were exposed to the test substance for 96 hours.

A range-finding study (phase I) was performed using the concentrations of 10 - 50 - 100 - 500 and 1,000 mg of test substance per litre of test medium (10 Brachydanio rerio per group). A control group (not exposed to any substance) was also employed under the same experimental conditions.

No mortality was recorded during the 96-hour study period, until 1,000 mg of test substance per litre of test medium. The range-fmding study was then considered as a limit test, and there was no need to perform an LC 50 evaluation above this concentration.

Any death and behavioural abnormalities (problems of swimming, equilibrium, pigmentation, fin morphology, ... ) were recorded approximately 2 hours, then 24, 48, 72 and 96 hours after the start of the study.

Measurements of pH, dissolved oxygen and temperature were performed at the start of the study for each test medium and the control, then approximately 24,48,72 and 96 hours later.

The study was performed in a static system (without renewal of test media during the study period).

Each test medium has been twice sampled (at the start and at the end of the study).

The quality criteria for the study as notified in the O.E.C.D. and E.E.C. protocols relative to the dissolved

oxygen concentration and mortality in the control group were respected.

Concentration levels of the test substance throughout the study period was guaranteed by the physicochemical

stability of the test substance in the experimental conditions.