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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:
2001-11-05 to 2002-03-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Remarks:
There were no amendents to the study plan.
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Remarks:
There were no amendents to the study plan.
GLP compliance:
yes (incl. certificate)
Remarks:
Certificate from the Swiss GLP Monitoring Authority
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 4.6, 10, 22, 46 and 100 mg/L
- Sampling method: no data
- Sample storage conditions before analysis: deep frozen at -20 °C and protected from light until analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: According to pre-experiments (without GLP) the test item was not sufficiently stable in test water. Therefore a semi-static test procedure was chosen: A flow-through test was not possible, since a concentrated homogeneous stock solution of T0021 02 could not be prepared.
- concentrated stock solution of nominal 100 mg/L was freshly prepared at the start of the test and before each test medium renewal by
dissolving 999 and 301 mg of the test item completely in 10 and 3 liters test water, respectively. For this, ultrasonic treatment for 15 minutes and stirring for 15 minutes at room temperature was applied. Adequate volumes of this intensively stirred stock solution were added to the test water in the aquaria and were intensively mixed to prepare the test media with the test concentrations as stated above.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebra fish
- Strain: Brachydanio rerio
- Source: Aquariumcenter Wildenauer, Munich, Germany
- Age at study initiation (mean and range, SD): no data
- Length at study initiation (length definition, mean, and SD): 3.1 +- 0.3 cm (Mean +- SD)
- Weight at study initiation (mean and range, SD): 0.32 +- 0.12 g (mean +- SD)
- Method of breeding: no data
- Feeding during test: no
- Food type: not applicable
- Amount: not applicable
- Frequency: not applicable

ACCLIMATION
- Acclimation period: 2 weeks
- Acclimation conditions (same as test or not): 1 week
- Type and amount of food: commercial fish diet (Tetra Min Hauptfutter, supplied by TETRA-Werke, D-49304 Melle, Germany)
- Feeding frequency: no data
- Health during acclimation (any mortality observed): no mortalities

QUARANTINE (wild caught)
- Duration: not applicable
- Health/mortality: not applicable
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
2.5 mmol/L (250 mg/L CaCO3)
Test temperature:
21-22 deg C
pH:
7.7-7.9
Dissolved oxygen:
7.8-8.5 mg/L. The test water was aerated prior to the start of the study until oxygen saturation was reached
Nominal and measured concentrations:
Control: 0 mg/L test substance
Nominal test item concentrations: 4.6, 10, 22, 46 and 100 mg/L test substance
Mean measured test item concentrations: not analysed, 8.7 mg/L (after 96h), 19.1 mg/L (after 96h), 38 mg/L (after 24h), 79.8 mg/L (after 24h).

In the analyzed test medium samples from the start of the test medium renewal periods the measured test item concentrations ranged between 88 and 100% of the nominal values. During the test medium renewal period of 24 hours a decrease of test item concentration in the test media was determined. At the end of the test medium renewal periods, 71 to 81 % of the nominal values were found. The losses may be due to instability of the test item in test water. The mean measured test concentrations (calculated as the average over all measurements per test concentration) varied in the range of 80 to 87% of the nominal values.
Details on test conditions:
TEST SYSTEM
- Test vessel: glass aquarium
- Material, size, headspace, fill volume: 5 L test volume
- Aeration: slight aeration during the test period
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7 fish were introduced in each aquarium in a random order

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: analytical grade salts
- Total organic carbon: no data
- Particulate matter: no data
- Metals: no data
- Pesticides: no data
- Chlorine: no data
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4 : 1 based on molarity
- Conductivity: no data
- Culture medium different from test medium: no
- Intervals of water quality measurement: once daily during the test in the freshly pepared and old test media

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: A 16-hour light to 8-hour darkness photoperiod, with a 30 minute transition period
- Light intensity: 50 to 500 Lux (light intensity at light period)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Geometric concentrations as reported in guideline requirements
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study no
- Test concentrations: not applicable
- Results used to determine the conditions for the definitive study: pretest for the stability of the test solution (RCC study No. 821801)
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
27 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Basis for effect:
mortality
Remarks on result:
other: 95% confidence interval 19-38.
Details on results:
- Behavior abnormalities: At the highest concentration, starting from 2h after exposure the fish were laying on side or back on the aquarium bottom, or were tumbling during the swimming. At 22 mg/L, starting 24h after exposure the fish were showing signs of appathy, staying at the bottom of the aquarium and fish was lying on side or back on the aquarium bottom.
- Observations on body length and weight: no data
- Other biological observations: no data
- Mortality of control: not applicable
- Other adverse effects control: not applicable
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
- Effect concentrations exceeding solubility of substance in test medium: no data
- 96 h LC0 = 19 mg/L (mean measured) (corresponding with a nominal concentration of 22 mg/L)
- 96 h LC100 = 38 mg/L (mean measured) (corresponding with a nominal concentration of 46 mg/L)
- 96 h NOEC = 8.7 mg/L (mean measured) (corresponding with a nominal concentration of 10 mg/L)
- 96 h LOEC = 19 mg/L (mean measured) (corresponding with a nominal concentration of 22 mg/L)
Reported statistics and error estimates:
The LC50 at the observation intervals 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 NOEC, LOEC, LC0 and LC100 were determined directly from the raw data.
Validity criteria fulfilled:
yes
Conclusions:
A semi-static acute toxicity test with zebra fish (Brachydanio rerio) performed with T002102 resulted in a mean measured 96-hour LC50 of 27 mg/L with a 95% confidence interval from 19 to 38 mg/L. Based on nominal concentrations, the 96-hour LC50 was 32 mg/L with a 95% confidence interval from 22 to 46 mg/L. The results of the test can be considered reliable without restriction.

Description of key information

In the guideline study (Piether, 2002), a semi-static acute toxicity test with zebra fish (Brachydanio rerio) was performed with T002102. The mean measured 96-hour LC50 of 27 mg/L was obtained with a 95% confidence interval from 19 to 38 mg/L. Based on nominal concentrations, the 96-hour LC50 was 32 mg/L with a 95% confidence interval from 22 to 46 mg/L. The results of the test can be considered reliable without restriction.

Key value for chemical safety assessment

LC50 for freshwater fish:
27 mg/L

Additional information