Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April 11, 1988 - April 22, 1988
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well described study, although not GLP, performed according to OECD 203, with the single deviation of no analytical measurements. This deviation does not invalidate the study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
No analytical confirmation of concentrations of test compound in test solutions.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Blown rapeseed oil 60 P.
No further information was presented in the report.

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solutions were prepared by mechanical dissolving (Ultra-Turrax) of the test substance in the required volume of deionised water. This stock solution was transferred into the test medium of one glass tank 2 hours before the start of the test (i.e. the addition of the fish, t=0 h). In the meantime, until the fish were transferred, the contents of each glass tank were mixed thoroughly using a mechanical stirrer in order to facilitate the dissolving and/or dispersing of the test sample.
- Eluate: not relevant
- Differential loading: not relevant
- Controls: controls were untreated
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not relevant
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)):not relevant
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The following was reported: "In the preparation of the different test batches …. It could be observed that charging the test medium with the test substance led to a flocky dispersion slowly settling up in the course of the test period.”

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Brachydanio rerio (zebra fish)
- Strain: -
- Source: West Aquarium GmbH & Co. KG, D-3422 Bad Lauterberg, Germany
- Age at study initiation (mean and range, SD): 2 months
- Length at study initiation (length definition, mean, range and SD): 2.0 ± 1.0 cm
- Weight at study initiation (mean and range, SD): no data
- Method of breeding: no data
- Feeding during test: unfed from 24 h prior to the start of the test
- Food type: not relevant
- Amount: not relevant
- Frequency: not relevant

ACCLIMATION
- Acclimation period: 13 days
- Acclimation conditions (same as test or not): yes
- Type and amount of food: Tetra Min® Hauptfutter
- Feeding frequency: twice daily until 24 hours before the test was started
- Health during acclimation (any mortality observed): no mortality

QUARANTINE (wild caught): not relevant
- Duration: not relevant
- Health/mortality: not relevant

Study design

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

Test conditions

Hardness:
Not measured during study
Test temperature:
Overall range during the test 20.8-23.8°C
pH:
Overall range during the test 7.7-7.9
Dissolved oxygen:
Overall range during the test >85% of saturation
Salinity:
Not relevant
Nominal and measured concentrations:
Nominal concentrations were 0, 100, 153, 244, 391, 625 and 1000 mg/L.
The nominal concentrations were not verified by chemical analysis.
Details on test conditions:
TEST SYSTEM
- Test vessel: glass tank
- Type (delete if not applicable): closed with glass plate allowing free air exchange
- Material, size, headspace, fill volume: glass, length 300 mm, width 200 mm, height 220 mm, volume 13.2 L, fill volume 10 L
- Aeration: yes, 1-3 bubbles per second
- Type of flow-through (e.g. peristaltic or proportional diluter): not relevant
- Renewal rate of test solution (frequency/flow rate): not relevant (static test)
- No. of organisms per vessel: ten
- No. of vessels per concentration (replicates): one
- No. of vessels per control (replicates): one
- No. of vessels per vehicle control (replicates): not relevant
- Biomass loading rate: no data

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water was prepared according to Annex 2 of OECD 203; deionised water used for its preparation was taken from the delivery system of the laboratory.
- 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
- Conductivity: no data
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH, oxygen and temperature were measured at 0, 1, 24, 48, 72 and 96 hours.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: no data
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality and fish behavior were observed at 0, 1, 24, 48, 72 and 96 hours.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5-1.6
- Justification for using less concentrations than requested by guideline: not relevant
- Range finding study: no
- Test concentrations: not relevant
- Results used to determine the conditions for the definitive study: not relevant
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
153 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Based on dispersions of the test material, no chemical analysis of test substance concentrations.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
625 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Based on dispersions of the test material, no chemical analysis of test substance concentrations.
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Based on dispersions of the test material, no chemical analysis of test substance concentrations.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Based on approximate solubility limit of test substance in test medium.
Details on results:
- Behavioural abnormalities: not reported.
- Observations on body length and weight: no data
- Other biological observations: no data
- Mortality of control: 0%
- Other adverse effects control: no data
- Abnormal responses: no data
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: see above.
- Effect concentrations exceeding solubility of substance in test medium: yes (see below)
Reported statistics and error estimates:
The reported LC50 (390 mg/L) was determined graphically by linear extrapolation using the mortalities of 153, 244, 391 and 1000 mg/L. This approach is considered to be inaccurate since the data point for 625 mg/L with 50% mortality is ignored. Based on graphical estimation with linear extrapolation between the data points, the LC50 is estimated to be 625 mg/L.

Any other information on results incl. tables

After 96 hours, mortality was 0% in the control and in the test solutions up to and including 153 mg/L, 40% at 244 and 391 mg/L, 50% at 625 mg/L and 100% at 1000 mg/L. It was not reported whether any sublethal effects were noted.

 

Evaluation.

The LC50 was 625 mg/L. The test solutions were dispersions, and the actual concentrations were not confirmed by chemical analysis. All tested concentrations were far above the solubility limit of the test material in water, which is <1 mg/L. The solubility limit in reconstituted test medium may be even less than that in water. Due to the fact that the test solutions were slowly settling dispersions far in excess of the solubility limit, the concentrations of which were not analytically determined, there is no information about the actual concentrations to which the test fish were exposed during the study. However, due to the method of preparation of the test solutions (thorough mixing of stock using Ultra-Turrax mixer, addition of stock to test water, followed by 2 hours of thorough mixing using a mechanical stirrer), it is sufficiently certain that the solubility limit of the test substance in the test water was achieved in all test solutions. Since at 100 -153 mg/L no fish mortality occurred, the conclusion of the study is that the LC50 is in excess of the solubility limit. The actual LC50 will be one or two orders of magnitude higher, based on the results of the present test. The mortality at 244 mg/L and above is likely to be due to physical effects of the test material (an oil), which above a certain threshold will for example impair the functioning of the gills.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
see above
Conclusions:
Fish acute toxicity test in Brachydanio rerio according to OECD 203: LC50 in excess of the solubility limit (hence in excess of about 1 mg/L); LC50 for exposure to dispersed test material 625 mg/L (no analytical verification of test substance concentrations).