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Ecotoxicological information

Short-term toxicity to fish

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short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
January 31, 2022
1 (reliable without restriction)
according to guideline
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
Specific details on test material used for the study:
The average log Kow varied from 5.4 to 6.0.
The water solubility was from 0.8 mg L-1 for the congener containing 4 Cl atoms to 0.3 mg L-1 for the congener with 9 Cl atoms.
Analytical monitoring:
Details on test solutions:
WAF preparation:
Loading rates (nominal concentrations): A stock elution was prepare using a nominal concentration of 1000 mg/L. After adding salts, it was then diluted to get all the tested concentrations.
Sample weight: Direct addition of 1000 mg (actial weight = 999.8 mg) per liter for the stock elution
Ambient temperature 20°C. Orbital shaker 100 rpm- 7 days.
At the end of shaking, the eluate is slightly turbid and whitish, partly jellified and hard to filter. After longer decantation a thin a whitish layer settles on the bottom of the container.
Filtration: With vacuum filtration apparatus and membranes with 0.45 µm pores of cellulose nitrate. After filtrations, the stock eluate was added with the salts and dilutions were prepared using the cultivation medium. Oxygenation of the eluate and its dilutions followed, via air insufflation for 30’ prior to starting the test and acclimatation to test temperature (20 °C).
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
The fishes came from a single group of similar length (1-2 cm) and age.
Test type:
Water media type:
Limit test:
Total exposure duration:
96 h
160 mg CaCO3/L
Test temperature:
23 °C
Dissolved oxygen:
≥ 60% (i.e. 5.14 mg/L at 23 °C and 1 atm)
0.13 ‰
Nominal and measured concentrations:
Limit test at 1000 mg/L
Key result
96 h
Dose descriptor:
Effect conc.:
> 0.8 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat. (dissolved fraction)
The WAF of different congeners C19Cl4 - C19Cl9 has been determined, the mean concentration was 0.8 mg/L. Specifically, the concentration in C19Cl4 was 0.9, in C19Cl5 0.07, in C19Cl6 0.03, in Cl19Cl7 0.02, in C19Cl8 0.02, in C19Cl9 0.03.
Basis for effect:
mortality (fish)
Key result
96 h
Dose descriptor:
Effect conc.:
> 1 000 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Validity criteria fulfilled:
After 96h of exposure at the limit concentration of 1000 mg/L, the sample does not induce the mortality of any of the seven Zebrafish under test (0/7), that is an acceptable value for the limit test. Overall, LC50 @ 96h > 1000 mg/L.
Executive summary:

This technical report summarizes all the data collected during the study: the exposure conditions of the fish at the tested sample limit concentration and the calculation of results. When using vertebrates for scientific purposes, as forced by the EU regulations, the 3 Rs rule must be obeyed. It includes three principles: Replacement, Reduction, and Refinement. Reduction refers to the use of methods that enable researchers to obtain comparable levels of information from fewer animals, or to obtain more information from the same number of animals. Whenever possible, for eco-tests on fishes, the limit test (one single concentration) is to be preferred to the serial concentrations test (at least five concentrations). Seven Zebrafishes, with a recommended total length of 1-2 cm, are exposed to the sample at the chosen limit concentration in breeding water, for a period of 96 hours. Mortality is recorded at 24, 48, 72 and 96 hours and compared with the negative control.

Description of key information

The substance has a nominal LC50 > 1000 mg/L. 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Dose descriptor:
Effect concentration:
> 1 000 mg/L

Additional information