Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2001-11-07 to 2002-02-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): T002113
- Physical state: pale yellow powder
- Analytical purity: 92.9%
- Lot/batch No.: RT002113G3A231
- Expiration date of the lot/batch: 2001-12-01
- Stability under test conditions: results from a hydrolysis pre-experiment (without GLP), indicated that, at a pH of 7, the test item concentration decreased rapidly within two hours by about 50% and thereafter stayed constant for at least five days (RCC Study No. 825265).
- Storage condition of test material: at room temperature at about 20°C away from direct sunlight.
- Other:
* solubility in water: 120 g/L
* pH in aqueous solution: 3.8 at a concentration of 120 g/L
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: RT00211G3A231
- Expiration date of the lot/batch: 2001-12-01 (retest date)
- Purity test date: no data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature at about 20°C
- Stability under test conditions: no data
- Solubility and stability of the test substance in the solvent/vehicle: no data

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Sampling method: duplicate samples were taken from the test media. The samples were acidified to about pH 3-5 immediately after sampling using 0.1 M phosphoric acid solution.
- Sample storage conditions before analysis: samples were analyzed immediately after sampling without prior storage.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a concentrated stock solution of nominal 100 mg/L is prepared at the start of the test by dissolving 1000 mg T002113 in 10 L test water by intense stirring for 5 minutes at room temperature. The lower test item concentrations were prepared by diluting the highest test concentration. The test media were freshly prepared just before introduction of the fish.
- Eluate: no
- Controls: yes

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Source: The test fish were obtained from Aquarium Center Wildenauer, Munich, Germany.
- Length at study initiation (length definition, mean, range and SD): 3.1 ± 0.3 cm
- Weight at study initiation (mean and range, SD): 0.32 ±0.12 g
- Feeding during test: no
- Food type: commercial diet fish (Tetra min Hauptfutter)

ACCLIMATION
- Acclimation period: one week prior the start to the same temperature and test water
- Acclimation conditions (same as test or not): yes
- Type and amount of food: commercial diet fish (Tetra min Hauptfutter)
- Health during acclimation (any mortality observed): no fish died and all fish were healthy

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
125 mg CaCO3/L
Test temperature:
22-23°C
pH:
7.3-7.9
Dissolved oxygen:
at least 8.0 mg/L or higher
Salinity:
not applicable
Conductivity:
not indicated
Nominal and measured concentrations:
nominal test concentrations: 4.6, 10, 22, 46, 100 mg/L
mean measured concentrations at the start of the test: 4.87, 10.4, 22.9, 46.1, 93.4 mg/L
mean measured concentrations after 2 days: 2.96, 6.49, 14.0, not applicable, not applicable mg/L
mean measured concentrations after 4 days: 2.88, 6.36, not applicable, not applicable, not applicable mg/L.

The mean measured concentrations (calculated as the average over all measurements per test concentration) ranged from 84 to 77% of nominal.
Details on test conditions:
TEST SYSTEM
- Test vessel: glass aquarium
- Material, size, headspace, fill volume: glass aquarium with 5 liter test medium
- Aeration: slightly aerated
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): not reported
- No. of vessels per control (replicates): not reported
- No. of vessels per vehicle control (replicates): not reported
- Biomass loading rate: lower than 1 g fish wet weight per liter test medium.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4 : 1 (base on molarity)
- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
- Photoperiod: 16-hour light to 8-hour darkness with a 30 minute transition period
- Light intensity: 50 to 500 lux

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 to 2.2
- Range finding study
- Test concentrations: data not shown
- Results used to determine the conditions for the definitive study: yes, range-finding test (without GLP)
Reference substance (positive control):
no

Results and discussion

Effect concentrations
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
12 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 7.8-19
Details on results:
- Behavioural abnormalities: apathy
- Mortality of control: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: foaming
- Effect concentrations exceeding solubility of substance in test medium: no remarkable observation, clear test medium.
- 96-h NOEC = 7.8 mg/L; 96-h LC100 = 19 mg/L
Reported statistics and error estimates:
- The LC50 and the 95% confidence interval at the observation dates could not be calculated by Probit Analysis or Moving Average Interpolation due to the steep concentration-effect relationship. Instead the LC50-values were determined as the geometric mean value of the two consecutive test concentrations with 0 and 100% mortality, and the 95% confidence intervals for the LC50 as the test concentrations with 0 and 100% mortality. The calculations of the LC values are based on the mean measured test item concentrations.
- The NOEC, LOEC, LCO and LC100 were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of T002113 to zebra fish was determined in a 96-hour static test according to the OECD 203 guideline. Under the conditions of the present study, the mean measured 96-h LC50 was determined to be 12 mg/L. The results of the test can be considered reliable without restriction.