Registration Dossier

Administrative data

short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
2 (reliable with restrictions)

Data source

Reference Type:
Bibliographic source:
not reported
Report date:

Materials and methods

Test guideline
equivalent or similar to guideline
OECD Guideline 203 (Fish, Acute Toxicity Test)
not specified
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:

Sampling and analysis

Analytical monitoring:
Details on sampling:

At the beginning and end of the exposure, measure the test substance concentrations at least in the lowest and highest test concentration groups. If it has been predicted that the test substance concentration decreases from the initial concentration by 20% or more during the exposure period, it is recommended to take measurements for all test concentration groups at the beginning and end of the exposure. Furthermore, for volatile or adsorptive substances or those that are likely to be greatly decreased during the exposure period, additional measurements are recommended at 24 hr intervals during the exposure period.

Study design

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

Test conditions

Details on test conditions:
-Test vessel: The test vessel or other instrument that contacts the test solution must be made of glass or other chemically inert materials. Use a test vessel of an appropriate size in regard to the recommended volume. Cap the vessel loosely to prevent evaporation and dust contamination.
If the test substance is volatile, perform the test in a sealed system. Use a sufficiently large vessel to prevent shortage of dissolved oxygen.

- test water: Use water suitable for culturing and testing the fish. It can be natural water (surface water or groundwater), dechlorinated tap water or artificially prepared water (see the note). The recommended total hardness is 10 - 250 mg/L in terms of calcium carbonate concentration, and the recommended pH is 6.0 - 8.5. Prepare the artificially prepared water using reagents of analytical grade and deionized or distilled water having an electric conductivity of 10 μS/cm or lower.

- Acclimatization: All test fish must be acquired at least 12 days before the test and acclimatized. After monitoring the fish for 48 hr, acclimatize them in water used for the test under the following conditions for at least 7 days before beginning the exposure. If possible, avoid using medicated bath after the monitoring period.
- Illumination: 12 - 16 hr light per day
- Temperature: Appropriate temperature for the species tested (see Appendix Table 1 under VI.)
- Oxygen concentration: At least 80% of the saturated oxygen concentration
- Feeding: Daily or three times per week until 24 hr before beginning the exposure.
- Record the mortality during the acclimatization period and apply the following conditions to the test fish.

If the overall mortality for seven consecutive days during the acclimatization period exceeds 10%, do not use them for the test.
If the overall mortality for seven consecutive days during the acclimatization period falls within 5 - 10%, extend the acclimatization period for 7 days.
If the overall mortality for seven consecutive days during the acclimatization period is below 5%, use them for the test.

- Test solution: To prepare a test solution of each concentration, directly dissolve the required amount of the test substance in the material water, or prepare a stock solution of the test substance at an appropriate concentration and dilute it with the material water. Follow the descriptions in "Preparation of the test solution" under "III. General rules."
Perform the test without adjusting the pH. If the pH of the material water is greatly changed by the addition of the test substance, it is recommended to perform an additional test after adjusting the pH to that observed prior to the addition of the test substance. Perform the pH adjustment through a method causing no change in the concentration, chemical reaction or precipitation of the test substance. Preferably, use HCl or NaOH for the pH adjustment.

- Test method: The test should be performed under a flow-through or semi-static condition. If the test substance concentration is unstable, a flow-through test is recommended.

- Volume: A maximum density of 1.0 g fish weight/L is recommended for a semi-static test. The volume can be larger in a flow-through test.
Number of test fish: Use at least 7 fish for each of the test concentrations and the control.

- Test concentrations: Adopt a concentration range comprising at least 5 concentrations that are setup in a geometric progression, preferably at a geometric ratio of within 2.2. The highest test concentration preferably gives fatal effects on all individuals tested, but concentrations of 100 mg/L or higher do not need to be tested. Preferably, no effect is observed at the lowest concentration.
Perform a control, and additionally an auxiliary control if using any auxiliary.
Reference substance (positive control):

Results and discussion

Effect concentrations
96 h
Dose descriptor:
Effect conc.:
ca. 6.6 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)

Applicant's summary and conclusion