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Short-term toxicity to aquatic invertebrates

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short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08/02/1993 - 04/05/1993
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study according to international guideline (OECD Guideline 202) under GLP , validity criteria met.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference Type:
study report
Report Date:

Materials and methods

Test guideline
according to
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Principles of method if other than guideline:
Not relevant
GLP compliance:
yes (incl. certificate)

Test material

Details on test material:
- Name of test material (as cited in study report): Vitamin A - Acetate
- Physical state: Solid
- Stability under test conditions: Good, tests are performed in total darkness, so test substance can not break down due to exposure to light.
- Storage condition of test material: At room temperature, in the dark
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not relevant

Sampling and analysis

Analytical monitoring:
Details on sampling:
- Concentrations:
Nominal: 10, 18, 32, 58 and 100 mg/l
Actual: 6.9, 9.2, 23.9, 53.4 and 106.7 mg/l
- Sampling method: Composite samples of each test concentration were drawn by mixing identical volumes of the test solutions taken from the approximate centers of the test vessels. they were taken immediately before exposure and after 48 hours exposure.
- Sample storage conditions before analysis: samples were kept at -18 to -22 degrees Celsius before analysis.

Test solutions

Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 1.00 g test substance and 1.01 g of a 96% 1-metyhl-2-pyrrolidone / 4% polyoxy-ethylene-sorbitan-monooleate mixture were mixed together (ultrasonic for 10 minutes). This blend was stored in the refrigerator (+4 degrees Celsius) for about 24 hours. 400 mg of this blend were mixed and made up to 2000 ml with water achieving a test substance concentration of 100 mg/l. Calculated amounts of the stock solution to produce the desired test concentrations were given into the water and were homogeneously distributed. the Daphnia were then transferred into the beakers.
- Controls: Yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): 1-metyhl-2-pyrrolidone / polyoxy-ethylene-sorbitan-monooleate
- Concentration of vehicle in test medium (stock solution and final test solution): 1-metyhl-2-pyrrolidone 96.0 mg/l; polyoxy-ethylene-sorbitan-monooleate: 4.0 mg/l
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Small parts of the test substance were swimming on the surface of the test water at 0 hour of exposure at all test concentrations and a deposit was observed at all test concentrations after 48 hours of exposure.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: daphnia magna
- Strain: Straus 1820
- Source: CIBA-GEIGY Ltd., testing facility
- Age at study initiation (mean and range, SD): 0-24 hours
- Length at study initiation (length definition, mean, range and SD): <800 um
- Method of breeding: Cultures of Daphnia are maintained in glass vessels containing approximately 2.5 l of reconstituted water at 20 +/- 1 degrees Celsius (water is renewed partially thrice weekly).
- Feeding during test No feeding during the test

- Acclimation period: 24 hours
- Acclimation conditions (same as test or not): yes, but test takes place in total darkness
- Type and amount of food: At each renewal the Daphnia are fed with a suspension of green algae (Scenedesmus subspicatus) supplemented by a suspension of TETRAMIN-extract in such quantities that the food is consumed after 24 h.
- Feeding frequency: Thrive weekly

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

240 mg/l CaCO3
Test temperature:
20 +/- 1 degrees Celsius
7.6 - 7.9, no change greater than 0.2 during test.
Dissolved oxygen:
37 - 99% of oxygen saturation value, no increase greater than 44% during the test.
Not relevant
Nominal and measured concentrations:
Nominal: 10, 18, 32, 58 and 100 mg/l
Measured (start): 11.3, 21.6, 45.3, 88.3 and 151.0 mg/l
Measured (end): 6.9, 9.2, 23.9, 53.4 and 106.7 mg/l
Details on test conditions:
- Test vessel: Beaker
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Glass beakers covered with watch glasses, 100 ml solution per beaker
- Aeration: Aerated with clean air for at least 24 hours before use, no aeration during test
- 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 containing 65 mg NaHCO3/l, 294 mg CaCl2 . 2 H2O/l, 123 mg MgSO4 . 7 H2O/l and 7 mg KCl/l

- Adjustment of pH: No
- Photoperiod: Total darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilization (every 24 hours)

- Spacing factor for test concentrations: 1.8
Reference substance (positive control):

Results and discussion

Effect concentrationsopen allclose all
48 h
Dose descriptor:
Effect conc.:
9.2 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
48 h
Dose descriptor:
Effect conc.:
107 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
48 h
Dose descriptor:
Effect conc.:
9.2 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
48 h
Dose descriptor:
Effect conc.:
46 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: 95% CL: 36-62 mg/l; Goodness t fit CHI: 3.13
Details on results:
- Behavioural abnormalities: None observed
- Observations on body length and weight: No data
- Other biological observations: No data
- Mortality of control: 0%
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No, substance concentrations at the end of the test were close to nominal concentrations.
- Effect concentrations exceeding solubility of substance in test medium: No
Results with reference substance (positive control):
Not relevant
Reported statistics and error estimates:
The EC50 values were calculated according to Berkson, JASA 48 (1953), 569-599. EC-values were graphically determined on gausso-logarithmic probability paper.

Any other information on results incl. tables

No data

Applicant's summary and conclusion

Validity criteria fulfilled:
Toxicity of Vitamin A Acetate towards Daphnia magna was investigated: the 48h-EC50 was found to be 46 mg/l, the 48h-NOEC was found to be 9.2 mg/l.
Executive summary:

Toxicity of Vitamin A Acetate towards Daphnia magna was investigated according to OECD Guideline 202 under GLP conditions. The 48h-EC50 was found to be 46 mg/l, the 48h-NOEC was found to be 9.2 mg/l. Test substance concentrations at the end of the experiment were close to nominal concentrations.