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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
yes
Remarks:
Acetone
Details on test solutions:
The stock solution 100 g/L was prepared by dissolving colourless liquid in acetone. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample in reconstituted water.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: Own breeding at University of Chemistry and Technology, Prague
- Age at study initiation (mean and range, SD): The animals used for the test shall be less than 24 h old and should not be first brood progeny
- Feeding during test: No feeding

ACCLIMATION - No data available
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Test temperature:
20±1°C
pH:
Test at 80 mg/l: 7.8 (no change during test)
Control: 7.9 (changed to 7.8 during test)
Contro + acetonel: 7.9 (changed to 7.8 during test)
Dissolved oxygen:
higher than 7.7 mg/L at the end of test
Nominal and measured concentrations:
0, 0, 5, 10, 20, 40, 80 mg/l nominal concentration
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 ml glass vessel
- fill volume: 25 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Natural water (surface or ground water), reconstituted water or dechlorinated tap water are acceptable as culturing and dilution water if D. magna survives in it for the duration of the culturing, acclimation and testing without showing signs of stress. Waters in the range pH 6 to pH 9, with hardness between 140 mg/l and 275 mg/l (as CaCO3) are recommended.
As an example, the preparation of dilution water meeting the requirements is described below.
Dissolve known quantities of reagents in water. The dilution water prepared shall have a pH of 7.8 ± 0.5, a hardness of (225 ± 50) mg/l (expressed as CaCO3), a molar Ca + Mg ratio close to 4 + 1 and a dissolved oxygen concentration above 7 mg/l.

Prepare the solutions specified below:
- Calcium chloride solution: Dissolve 117.6 g of calcium chloride dihydrate (CaCl2.2H2O) in water (4.2) and make up to 1 l with water (4.2).
- Magnesium sulfate solution: Dissolve 49.3 g of magnesium sulfate heptahydrate (MgSO4.7H2O) in water (4.2) and make up to 1 l with water (4.2).
- Sodium bicarbonate solution: Dissolve 25.9 g of sodium bicarbonate (NaHCO3) in water (4.2) and make up to 1 l with water (4.2).
- Potassium chloride solution: Dissolve 2.3 g of potassium chloride (KCI) in water (4.2) and make up to 1 l with water (4.2).

Mixing
Mix 2.5 ml of each of the four solutions and make up to 1 l with water.
The dilution water shall be aerated until the dissolved oxygen concentration has reached saturation and the pH has stabilized. If necessary, adjust the pH to 7.8 ± 0.5 by adding sodium hydroxide (NaOH) solution or hydrochloric acid (HCI). The dilution water prepared in this way shall not be further aerated before use.

- Sodium hydroxide solution, e.g. [NaOH] : 1 mol/l.
- Hydrochloric acid, e.g. [HCl] : 1 mol/l.

Reference substance:
Dissolve 600 mg of potassium dichromate (K2Cr2O7) in water and make up to 1 l with water (4.2).

OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment done
- Photoperiod: No - Darkness
- Light intensity:

CALCULATION:
EC50 was calculated using non linear regression by the software Prism 4.0
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2Cr2O7)
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
28.9 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95 % CI was 24.3 - 34.4 mg/l
Results with reference substance (positive control):
- Results with reference substance valid
- EC50: 0.79 mg/L (24 hours)
Reported statistics and error estimates:
EC50 was calculated using non linear regression by the software Prism 4.0
Validity criteria fulfilled:
yes
Conclusions:
The median effective concentration (EC50) for the test substance, in Daphnia magna was determined to be 28.9 mg/L on the basis of immobility in a 48 hour study. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic chronic 3 category as per the CLP classification criteria.
Executive summary:

Aim of this study was to determine the toxicity of test chemical on the aquatic invertebrates, daphnia magna by performing the test according to the OECD guideline 202. Test was conducted under the static system for the total exposure period of 48 hrs.

The stock solution 100 g/L was prepared by dissolving colourless liquid in acetone. Test solutions of required concentration as were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0, 5, 10, 20, 40, 80 mg/l nominal concentrations were used. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. The median effective concentration (EC50) for the test substance, on Daphnia magna was determined to be 28.9 mg/L on the basis of immobility after 48 hours. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic chronic 3 category as per the CLP classification criteria.

Description of key information

The median effective concentration (EC50) for the test substance, in Daphnia magna was determined to be 28.9 mg/L on the basis of immobility in a 48 hours study.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
28.9 mg/L

Additional information

Experimental key study and supporting predicted data for the test chemical and supporting studies for its structurally similar read across substance were reviewed for the short-term toxicity to aquatic invertebrates which are summarized as below:

 

Aim of this study was to determine the toxicity of test chemical on the aquatic invertebrates, daphnia magna by performing the test according to the OECD guideline 202. Test was conducted under the static system for the total exposure period of 48 hrs.

The stock solution 100 g/L was prepared by dissolving colourless liquid in acetone. Test solutions of required concentration as were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0, 5, 10, 20, 40, 80 mg/l nominal concentrations were used. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. The median effective concentration (EC50) for the test substance, on Daphnia magna was determined to be 28.9 mg/L on the basis of immobility after 48 hours. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic chronic 3 category as per the CLP classification criteria.

 

Further, Estimation Programs Interface Suite (EPI suite) was run to predict the toxicity of the test compound to daphnia magna. The LC50 value of the test chemical, for 48 hours, on daphnia magna was estimated to be 14.167mg/L.

Based on the immobilization of test animal daphnia magna for 48 hrs of exposure, the chemical was considered to be toxic and can be classified as aquatic chronic 3 as per the CLP classification criteria.

Study was conducted according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test), to determine the effect of test chemical on Daphnia magna. The test substance was soluble in Adams media. Therefore, the test solution was prepared by dissolving 100 mg of the test substance in 100 ml of ADaM’s media. Achieving test concentrations of 3.125 mg/L,6.25 mg/L,12.5 mg/L,25 mg/L, and 50 mg/L, respectively, the test Daphnia magna were exposed to these concentration for 48 hours. The experimental median effective Concentrations [EC-50 (48 h)] for test chemical on daphnia was observed to be 50 mg/l. Thus, based on the immobilization of test animal daphnia magna after 48 hrs of exposure, chemical was considered to be toxic and was classified as aquatic chronic 3 as per the CLP classification criteria.

Thus, based on the above observation, the test chemical was considered to be toxic and can be classified as aquatic chronic 3 as per the CLP classification criteria.