Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2005-01-10 to 2005-03-31
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant; Guideline Study (OECD 203)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Sampling method: In this semi-static test with a test medium renewal every day, fourfold samples were taken from the freshly prepared test medium and the control at the start of the test (Day 0) and at the last test medium preparation on Day 3. Additionally, fourfold samples were taken from the test medium and the control at the end of the first test medium renewal period (Day 1) and at the end of the last test rnediumrenewal (Day 4).
- Sample storage conditions before analysis: The samples were stored deep-frozen and protected from light until analysis was performed.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test medium with a loading rate of nominal 100 mg/L was prepared prior to the start of the test and prior to each test medium renewal by weighing nominal 500 mg of the test item into 5000 mL of test water. No auxiliary solvent or emulsifier was used. The test item was mixed into the test water as homogeneously as possible using ultrasonic treatment for 15 minutes and intense stirring for 96 hours at room temperature in the dark to dissolve a maximum concentration of the test item in the dispersion. The long stirring period of 96 hours was chosen according to the results of a pre-test (without GLP) which showed that during this time no significant loss of the test item was to be expected. Then, the supersaturated stock dispersion of the test item was filtered through a membrane filter (Schleicher & Schuell, Type NC45, pore size 0.45 pm) after the 96 hours stirring period just before the preparation of the test medium. The undiluted filtrate with the maximum concentration of dissolved test item was used as test medium.
- Differential loading: 100 mg/L
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): No auxiliary solvent or emulsifier was used.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebra fish (Brachydanio rerio)
- Source: supplier H. Eggimann, CH-4133 Prattein, Switzerland
- Length at study initiation: 2.8 ± 0.22 cm (Mean ± SD)
- Weight at study initiation: 0.18 ± 0.04 g (Mean ±SD)
- Feeding during test: Fish were not fed during the test

ACCLIMATION
- Acclimation period: 1 week
- Acclimation conditions (same as test or not): yes
- Type and amount of food: commercial fish diet


Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
125 mg/L as CaCO3;
Test temperature:
21 - 22 °C;
pH:
7.1 - 7.3;
Dissolved oxygen:
8.4 - 9.0 mg/L;
Salinity:
alkalinity: 0.4 mmol/L;
Nominal and measured concentrations:
Nominal loading rate: 100 mg/L
Measured: 0.80 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: One glass aquarium with five liters test medium was used for each treatment (the single test concentration and the control)
- Renewal rate of test solution: daily
- No. of organisms per vessel: 7 fish per vessel

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water according to the Commission Directive 92/69/EEC and the OECD Guideline.

OTHER TEST CONDITIONS
- Adjustment of pH: 7.1 - 7.3
- Photoperiod: Photoperiod of 16 hours light and 8 hours darkness (with a 30 minute transition period). The test was performed under reduced light conditions to avoid photolytic degradation of the test item (< 50 LUX)


TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: According to the results of a pre-experiment (without GLP) the test item was not soluble at a concentration of 100 mg/L in test water and no homogeneous dispersion could be prepared.
A limit test was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effect on the test fish up to the highest concentration which could be dissolved in the test water at a loading rate of 100 mg/L. Additionally, a control (test water without test item) was tested in parallel.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: masured: 0.80 mg/L
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: measured: 0.80 mg/L
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: measured: 0.80 mg/L
Details on results:
- Behavioural abnormalities: None observed
- Mortality of control: None;

Any other information on results incl. tables

Since WAFs were prepared, all reported biological results are related to the loading rate of 100 mg/L of the test item.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
With high probability not acutely harmful to fish. No toxic effects within the range of solubility.