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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:
1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
1992
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: all test concentrations and the control
- Sample storage conditions before analysis: In pre-experiments significant losses of test article were determined in deep-frozen test medium samples. Therefore, all samples were analyzed immediately after sampling without prior storage.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
The test medium of the highest test concentration of nominal 100 mg/L was prepared by dissolving 1.6 g of the test article in 10 liters test water by intense stirring and short ultrasonic treatment. The amount of test article required was adjusted based on its purity (62.2%).
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebra fish
- Source: Zoohaus Schaub, CH-4410 Liestal, Switzerland
- Length at study initiation: 3.2 ± 0.2 cm (Mean ± SD)
- Weight at study initiation: 0.3 ± 0.1 g (Mean ± SD)

ACCLIMATION
- Acclimation period: one week
- Acclimation conditions: same as test conditions
- Type and amount of food during acclimation: commercial fish diet (TETRA MIN Hauptfutter, supplied by TETRA-Werke, D-49304 Melle, Germany)
- Feeding frequency during acclimation: not specified
- Health during acclimation: no mortality

FEEDING DURING TEST
no feeding during test
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
21 - 22 °C
pH:
7.4 - 8.2
Dissolved oxygen:
8.0 - 8.3 mg/L
Nominal and measured concentrations:
Nominal concentrations: 4.6, 10, 22, 46, 100 mg/L
Measured concentrations: 3.2, 6.7, 15.6, 31, 73 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: aquarium
- Material, fill volume: glass, 5 L
- Aeration: slightly aerated
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST WATER PARAMETERS
- Preparation of dilution water: analytical grade salts were added to purified water to obtain the following nominal concentrations:
CaCI2 x 2H20: 2.0 mmol/L (= 294.0 mg/L)
MgS04 x 7H20: 0.5 mmol/L (= 123.0 mg/L)
NaHC03: 0.75 mmol/L (= 65.0 mg/L)
KCI: 0.075 mmol/L (= 5.8 mg/L)
Water Hardness Alkalinity: 2.5 mmol/L (= 250.0 mg/L) as CaC03 0.8 mmol/L
Ratio of Ca : Mg = 4 :1 (based on molarity)
Na : K =10:1 (based on molarity)

OTHER TEST CONDITIONS
- Adjustment of pH: yes, for all concentrations except 4.6 mg/L adjustment with 0.1M (NaOH) to values between 7.5 - 7.7.
- Photoperiod: 16-hour light to 8-hour darkness
- Light intensity: 200 - 1200 Lux

EFFECT PARAMETERS MEASURED
The test fish were observed after approximately 3, 24, 48, 72 and 96 hours test duration for symptoms of intoxication and mortality.
The water temperature, pH-values and dissolved oxygen concentrations in the test media of all test concentrations and the control were measured before the start of the test and once every day during the test. The appearance of the test media was recorded at the beginning of the test and once every day during the test.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Range finding study
- Test concentrations: not specified
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 73 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
73 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
No mortalities observed
- Behavioural abnormalities: Slight apathy at the highest concentration (73 mg/L measured), starting at 24h
- Observations on body length and weight: no
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations that might cause a difference between measured and nominal values: no
Validity criteria fulfilled:
yes
Conclusions:
According to OECD 203 the test item showed a LC50 >73 mg/L.
Executive summary:

The acute toxicity of the test substance was evaluated with Zebrafish using a 96-hour static test according to OECD guideline No. 203 and EU method C.1. A preliminary test preceded the definitive test. Five concentrations of the test substance were used together with a control at: 4.6, 10, 22, 46 and 100 mg/L. 14 daphnids (in two replicates of seven) were exposed to each concentration for 96 hours and mortality and symptoms of intoxications were recorded at 3, 24, 48, 72 and 96 hours. Initial concentrations of the test item measured in the definitive test solutions were 67% till 73 % of the corresponding nominal values. Therefore, the reported biological results are related to the total mean measured test article concentrations: 3.2, 6.7, 15.6, 31 and 73 mg/L. Under the experimental conditions, the 96-hour LC50 of the test substance in a static test system is >73 mg/L for Zebrafish.

Description of key information

According to OECD 203 the test item showed a LC50 >73 mg/L.

Key value for chemical safety assessment

Additional information

The acute toxicity of the test substance was evaluated with Zebrafish using a 96-hour static test according to OECD guideline No. 203 and EU method C.1. A preliminary test preceded the definitive test. Five concentrations of the test substance were used together with a control at: 4.6, 10, 22, 46 and 100 mg/L. 14 daphnids (in two replicates of seven) were exposed to each concentration for 96 hours and mortality and symptoms of intoxications were recorded at 3, 24, 48, 72 and 96 hours. Initial concentrations of the test item measured in the definitive test solutions were 67% till 73 % of the corresponding nominal values. Therefore, the reported biological results are related to the total mean measured test article concentrations: 3.2, 6.7, 15.6, 31 and 73 mg/L. Under the experimental conditions, the 96-hour LC50 of the test substance in a static test system is >73 mg/L for Zebrafish.