Retinyl acetate

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08/02/1993 - 04/05/1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study according to international guideline (OECD Guideline 202) under GLP , validity criteria met.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Principles of method if other than guideline:

Not relevant

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Retinyl acetate

EC Number:

204-844-2

EC Name:

Retinyl acetate

Cas Number:

127-47-9

Molecular formula:

C22H32O2

IUPAC Name:

(2E,4E,6E,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohex-1-en-1-yl)nona-2,4,6,8-tetraen-1-yl acetate

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:

yes

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

Vehicle:

yes

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:

TEST ORGANISM
- 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
- 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:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation period

Test conditions

Hardness:

240 mg/l CaCO3

Test temperature:

20 +/- 1 degrees Celsius

pH:

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.

Salinity:

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 SYSTEM
- 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



TEST MEDIUM / WATER PARAMETERS
- 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


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Total darkness


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


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

9.2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

107 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

9.2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

46 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

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:

yes

Conclusions:

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.