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Ecotoxicological information

Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The zebra fish were exposed to the test substance from 18 to 22 December 2006. The test solutions were analysed on 22 December 2006.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Only two concentrations were tested and therefore the LC50 value was not determined.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
one adaptation : the test was carried out by testing the lowest EC50 value found in algea or in daphnia and at a 10-fold higher concentration instead of a least five concentrations in a geometric series.
Deviations:
yes
Remarks:
The fish were fed on the day of test initiation. To the opinion of the study director these deviations did not influence the integrity of the study.
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
To study the exposure of the fish to F1EC, samples of 10 mL were taken in duplicate from all test solutions on day 0 and on day 4.
Vehicle:
no
Details on test solutions:
The test was carried out by testing the lowest EC50 value found in algae or in daphnia and at a 10-fold higher concentration. The lowest EC50 value of 6 mg/L was found during the algae test (De Groot, 2006). Based on these results and the results of a preliminary test the study was conducted as a limit test at F1EC concentrations of 0, 6 and 60 mg/L.
On the day of test initiation a stock solution of 60.3 mg/L was prepared by dissolving 301.7 mg F1EC in 5 litre ISO water. Test solutions were prepared at test initiation by dilution of the stock solution in ISO water. One aquarium (lenght, width and height of 30, 20 and 15 cm respectively) was prepared for each concentration with 4 litre test solution each containing 8 zebra fish. The fish were exposed to the test solutions during 96 hours. All test solutions were clear and particles of the test substance were not visible.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
Zebra fish used in this test originate from a commercial supplier (De Rifwachter, Hilversum, The Netherlands). The fish arrived at the laboratory of the HES Department on 1 December 2006. The fish are kept in a climate chamber with a temperature setpoint of 23°C in a plastic container with 100 litre ISO-water. The light regime is 14 hours light and 10 hours dark (light on from 8 am to 10 pm). The fish are fed with Tetramin and living water fleas. The water is filtered continuously to assure an acceptable water quality.
No aberrant behaviour or appearance of the fish was observed on the day of test initiation. The total weight of 10 fish was determinaed on the day of test initiation. The total weight of the 10 fish was 3.60 gram which resulted in a mean weight of 0.360 gram per fish. Based on these results the weight of the fish per litre test solution was lower than 1 g per litre.
Test type:
static
Water media type:
freshwater
Total exposure duration:
96 h
Test temperature:
23°C +/- 2°C.
The measured temperature of the thermostated climate chamber ranged between 23.0°C and 23.8°C during the test.
pH:
At test initiation the pH of the ISO water was 7.4 and the pH of the stock solution was 7.2
At test end, the pH was 8.2 in the control, 7.8 at 6 mg/L and 6.7 at 60 mg/L.
Dissolved oxygen:
At test initiation the oxygen content of the ISO water and the stock solution was 8.7 mg/L.
At test end, the the oxygen content was 7.9 mg/L in the control, 8 mg/L at the test solution of 6 mg/L and 8.7 mg/L at the test solution of 60 mg/L.
Nominal and measured concentrations:
Nominal concentrations were 6 and 60 mg/L, measured concentrations at t0 were 5.8 and 64.2 mg/L respectiveley, measured concentration at day 4 was 6.3 mg/L for the nominal concentration of 6 mg/L. For the nominal concentration of 60 mg/L, no measurement was performed at day 4 since all fish died within 3 hours after test initiation.
Details on test conditions:
Environmental conditions:
8 zebra fish were added in a randomized way to all aquariums. The aquariums were covered with plastic plates. The test solutions were continuously aerated during the test.
The aquariums were placed in a climate chamber with a temperature setpoint of 23°C. The test temperature should be between 21 and 25°C and should be constant within a range of 2°C. The air temperature was recorded once every hour.
The light regime was 14 hours light and 10 hours dark.

Biological observations and measurements:
At 3, 24, 48, 72 and 96 hours after test initiation the behaviour and appearance of the surviving fish was recorded. At the end of the test the surviving fish were killed with tricainesulfonate. The total lenght (tail included) of each fish was determined.
On day 0 the pH, oxygen content and temperature of the ISO-water and the stock solution were measured. At test termination the water parameters were measured in all aquariums.

Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 6 - < 60 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
60 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
No mortality was observed in the control aquarium.
The results of the biological observations are given in Table 1.
Because neither mortality nor effects on behaviour and general appearance were observed at a nominal concentration of 6.0 mg/l, this concentration is considered a No Observed Effect Concentration (NOEC).
All fish at 60 mg/l were found dead within 3 hours after test initiation. The LC50 value based on nominal concentrations was established to be within the range of 6.0 - 60 mg/l.

At the end of the study, the fish were killed with a solution of 2 g tricainesulfonate (Ethyl 4-aminobenzoate) and 2 g sodium bicarbonate and the total length (tail included) of each fish was determined. The mean length of the zebra fish was 3.0 cm. The mean length of the zebra fish was in agreement with the recommended fish length ofOECD Guideline 203 of 2.0 ± 1.0 cm.
Sublethal observations / clinical signs:

 Table 1 : Survival of zebra fish during the test

 Nominal concentration of F1EC

(mg/L)

 Number of survining fish            

 Mortality at the end of the test

(%)

   3 hours  24 hours  48 hours  72 hours  96 hours  
 0  8  8  8  8  8  0
 6  8  8  8  8  8  0
 60  0  0  0  0  0  100
Validity criteria fulfilled:
yes
Remarks:
There was no mortality in the control (< 10%), constant conditions were maintained throughout the test, the O2 concentration was at least 60 % of the air saturation value; the concentration of the hydrolysed substance has been satisfactorily maintained.
Conclusions:
The 96h LC50 value of F1EC for Danio rerio has been determined to be between 6 and 60 mg/L.
Executive summary:

The acute toxicity of F1EC (Monofluoroethylene carbonate) to zebra fish (Danio rerio) was tested following OECD Guideline 203 (OECD, 1992) and according to GLP.

 

The test was carried out by testing the lowest EC50 value found in algae or in daphnia and at a 10-fold higher concentration. The lowest EC50 value of 6 mg/L was found during the algae test (De Groot, 2006). Based on these results and the results of a preliminary test the study was conducted as limit test at F1EC concentrations of 0, 6.0 and 60 mg/L.

 

One aquarium was prepared per test concentration containing 4 liter test solution and 8 zebra fish. The test was terminated after 96 hours of exposure.

 

During a test with Daphnia magna (De Groot, 2005) it was demonstrated that F1EC was almost completely hydrolysed already at test initiation. For this reason it is not possible to measure the concentration of F1EC with a specific analysis and it was decided to determine the concentration F1EC based on the fluoride content of the samples with cuvette test. F1EC contains 18% fluoride based on the molecular formula. Based on the analytical results, the difference between the nominal and the mean measured concentrations was < 20% and for this reason the endpoints of the test were based on nominal concentrations.

 

No mortality was observed in the control aquarium. Because neither mortality nor effects on behaviour and general appearance were observed at a nominal concentration of 6.0 mg/L, this concentration is considered as No Observed Effect Concentration (NOEC). All fish at 60 mg/L were found dead within 3 hours after test initiation.

The 96h LC50 value based on nominal concentrations was established to be within the range of 6.0 - 60 mg/L.

Description of key information

The test was carried out by testing the lowest EC50 value found in algae or in daphnia and at a 10-fold higher concentration. The lowest EC50 value of 6 mg/L was found during the algae test. Based on these results and the results of a preliminary test the study was conducted as limit test at concentrations of 0, 6.0 and 60 mg/L.

The 96h LC50 value of the substance for Danio rerio has been determined to be between 6 and 60 mg/L. At 6 mg/L no mortality occurred and at 60 mg/L 100% mortality occurred. Based on this the EC50 value is conservatively estimated to be 30 mg/L (half of the concentration that caused 100% mortality). No data on marine species is available.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
30 mg/L

Additional information