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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10/05/1995 - 14/06/1995
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
other: published parts of study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
Test material form:
liquid: viscous
Specific details on test material used for the study:
No test surrogate or analogue material was used.

Sampling and analysis

Analytical monitoring:
Details on sampling:
- Concentrations: nominal concentrations of 5.8, 10, 18, 32, 58 and 100 mg/l
- Sampling method: Water samples of each concentration were taken at 0 and 48 hour exposure and analyzed for the content of d,l-a-tocopherol. The 0 hour samples of each test concentration (2 x 150 ml volume each) were taken from one additional beaker (600 ml volume), which equilibrated in parallel to the original test beakers of each concentration during a time of 24 hours before test initiation. Additionally one sample was taken from the freshly prepared stock solution (50 ml). At test termination (48 hours), samples of about 80 ml were taken from three of the four replicate test vessels of each treatment group, of which at least one sample was analyzed.

Test solutions

Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 200 mg test substance were added to 8 ml Tween 80 (0.1% w/v) and about 200 ml reconstituted water (M4-medium according to elendt)
and mixed for 1 minute with a Polytron-mixer. This dilution was filled up to 2000 ml with M4-medium (according to Elendt). Calculated amounts of
this stock solution were transferred to 1000 ml test medium and homogeneously distributed to produce the desired test concentrations.
- Controls: bare M4 medium
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Tween 80 (0.1%)
- Concentration of vehicle in test medium (stock solution and final test solution): Amounts of 3.77, 3.6, 3.28, 2.72, 1.68 and 0 ml per litre were added to the test concentrations of 5.8, 10, 18, 32, 58 and 100 mg d,l-a-tocopherol per litre, respectively.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): d,l-a-Tocopherol built a separate layer on the water surface after dilution with
the test medium

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: Daphnia magna STRAUS
- Strain: Clon 5
- Source: In-house culture of CIBA-GEIGY Limited
- Age at study initiation (mean and range, SD): <24 hours
- Method of breeding: In-house cultures of Daphnids were maintained in polypropylene vessels containing approximately 2.5 l of M4 medium. About one third of this culture medium was renewed three times a week. The daphnids were fed daily with a suspension of green algae (1.0 ml/L of a
Scenedesmus subspicatus suspension with a density of approx. 10^8 cells/ml). Cultering was performed under constant conditions. Three times a
week the young daphnids were separated to maintain a homogenous stock of animals.

- Acclimation period: +/- 24 hours
- Acclimation conditions (same as test or not): 16/8h light/dark conditions, while test is carried out in the dark entirely.

Study design

Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
Post exposure observation period:
No post exposure observation period.

Test conditions

Test temperature:
20 +/- 2 °C
7.4 - 8.1, maximum increase of 0.6 during test
Dissolved oxygen:
0h: 97 - 100%
48h: 94 - 98%
Nominal and measured concentrations:
Nominal: 5.8, 10, 18, 32, 58 and 100 mg/l
Measured (0h): 4.9(4.7/5.0), 4.5 (4.5/4.5), 7.9 (7.8/8.1), 13.1 (13.1/13.0), 21.2 (20.0/22.3) and 30.3 (27.4/33.2) mg/l
Measured (48h): 2.0 (1.9/2.1), 2. (2.2/2.1), 4.0 (3.8/4.1), 5.8 (5.7/5.9), 9.9 (9.9/9.8) and 16.8 (16.1/17.4) mg/l
Effect concentrations were determined on the base of nominal concentrations.
Details on test conditions:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass (150 ml, with 100 ml test solution)
- Aeration: paasive via diffusion
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): 4

- Source/preparation of dilution water: Reconstituted water
- Culture medium different from test medium: No
- Intervals of water quality measurement: 48h

- Adjustment of pH: No
- Photoperiod: Test was carried out in the dark

- Spacing factor for test concentrations: 1.8
- Range finding study
- Test concentrations: 0.01, 0.1, 1, 10 and 100 mg/l
- Results used to determine the conditions for the definitive study: Yes
Reference substance (positive control):

Results and discussion

Effect concentrationsopen allclose all
Key result
48 h
Dose descriptor:
Effect conc.:
> 23.53 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: nominal > 100 mg/L
Key result
48 h
Dose descriptor:
Effect conc.:
9.43 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: nominal 32 mg/L
Details on results:
- Behavioural abnormalities: no abnormalities reported
- Other biological observations: No unexpected phenomena reported
- Mortality of control: less than 10%
- Abnormal responses: None reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: d,1-a-Tocopherol built a separate layer
on the water surface after dilution with the test medium. The substance also stuck all over the daphnids.
- Effect concentrations exceeding solubility of substance in test medium: No
Results with reference substance (positive control):
No reference substance used.
Reported statistics and error estimates:
No data on statistics, errors or confidence intervals available.

Applicant's summary and conclusion

Validity criteria fulfilled:
no mortality in control
The 48h EC50 of d,l-alpha-Tocopherol to Daphnia magna was found to be > 23.53 mg/l, 48h; NOEC was found to be 9.43 mg/l
Executive summary:

The toxicity of d,l-alpha-Tocopherol towards Daphnia magna was determined according to OECD Guideline 202 under GLP. Exposure concentrations decreased during exposure period, therefore concentrations are expressed on average measured concentrations. The 48h EC50 of d,l-a-Tocopherol to Daphnia magna was found to be > 23.53 mg/l, 48h NOEC was found to be 32 mg/l.