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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:
Experimental test result performed using standard test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Principles of method if other than guideline:
Short term toxicity of test chemical to aquatic invertebrates was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system.
GLP compliance:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): isopropyl 4-hydroxybenzoate
- Molecular formula : C10H12O3
- Molecular weight : 180.202 g/mol
- Smiles notation : c1(C(OC(C)C)=O)ccc(O)cc1
- InChl : 1S/C10H12O3/c1-7(2)13-10(12)8-3-5-9(11)6-4-8/h3-7,11H,1-2H3
- Substance type:Organic
- Physical state:Solid
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
The stock solution 100 mg/l was prepared by dissolving white powder in reconstituted water. The solution was kept in ultrasonic bath for 20 min. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test 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
- Acclimation period:
- Acclimation conditions (same as test or not):
- Type and amount of food:
- Feeding frequency:
- Health during acclimation (any mortality observed):
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Remarks on exposure duration:
± 1 hr
Test temperature:
20±1°C
pH:
Sample at concentration 16 mg/l: 7.6 changes to 7.8 during test
Control: 7.7 changes to 7.9 during test
Dissolved oxygen:
higher than 7.9 mg/L at the end of test
Nominal and measured concentrations:
0, 0.5, 1, 2, 4, 8 and 16 mg/l
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:
72 h
Dose descriptor:
EC50
Effect conc.:
3.79 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95 % CI was 3.07 to 4.40 mg/l
Results with reference substance (positive control):
- Results with reference substance valid
- EC50: 0.73 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 isopropyl 4-hydroxybenzoate in Daphnia magna was determined to be 3.79 mg/L on the basis of mobility inhibition effects in a 48 hour study.
Executive summary:

Aim of this study was to assess the short term toxicity of isopropyl 4-hydroxybenzoate to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs.

 

The stock solution 100 mg/l was prepared by dissolving white powder in reconstituted water. The solution was kept in ultrasonic bath for 20 min. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0.5, 1, 2, 4, 8 and 16 mg/l concentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0.

 

The median effective concentration (EC50) for the test substance isopropyl 4-hydroxybenzoate in Daphnia magna was determined to be 3.79 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be hazardous to aquatic invertebrate but as the chemical was readily biodegradable in water thus on that criteria, chemical consider to be nontoxic and not classified as per the CLP classification criteria.

Description of key information

Aim of this study first experimental key was to assess the short term toxicity of isopropyl 4-hydroxybenzoate to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs. The stock solution 100 mg/l was prepared by dissolving white powder in reconstituted water. The solution was kept in ultrasonic bath for 20 min. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0.5, 1, 2, 4, 8 and 16 mg/l concentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance isopropyl 4-hydroxybenzoate in Daphnia magna was determined to be 3.79 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be hazardous to aquatic invertebrate but as the chemical was readily biodegradable in water thus on that criteria, chemical consider to be nontoxic and not classified as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
3.79 mg/L

Additional information

In different studies, the given test chemical has been investigated for toxicity of chemical on mobility of aquatic invertebrates to a greater or lesser extent. The studies are summarized as below –

 

Aim of this study first experimental key was to assess the short term toxicity of isopropyl 4-hydroxybenzoate to aquatic invertebrates daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) in a static system for the total exposure period of 48 hrs. The stock solution 100 mg/l was prepared by dissolving white powder in reconstituted water. The solution was kept in ultrasonic bath for 20 min. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0.5, 1, 2, 4, 8 and 16 mg/l concentrations were used in the study. Effects on immobilisation were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, effect concentration EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance isopropyl 4-hydroxybenzoate in Daphnia magna was determined to be 3.79 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be hazardous to aquatic invertebrate but as the chemical was readily biodegradable in water thus on that criteria, chemical consider to be nontoxic and not classified as per the CLP classification criteria.

 

Above study was supported by the second study from peer reviewed journal. Acute immobilization test was performed in Daphnia magna was performed at different concentrations of isopropylparaben for 24- 48 hrs according to the OECD TG 202.Four groups of five organisms were used in 100 ml of glass beaker. Number of immobilized daphnids was checked after 24 and 48 hrs. After experiment it was found that, the EC50 value for short term toxicity to Daphnia magna for isopropylparaben (4191-73-5) was determined to be 30 mg/l concentration. Based on the value, the isopropylparaben was considered to be toxic to aquatic invertebrates but as it was readily biodegradable hence, it can be considered to be not classified as per the CLP regulations.

Similarly short-term toxicity to daphnia magna was performed for 48 hrs at five nominal concentrations in a temperature-controlled incubator at 24°C with a 16:8 light: dark cycle.100 ml beaker was taken having 80 ml of test chemical. Test solution was prepared by dissolved in high-performance liquid chromatography grade acetone and delivered into RHW at 0.1% acetone. No aeration was used and LC50 value was determined at 24 and 48 hrs. On the basis of above experiment result the LD 50 value for Daphnia magna for isopropylparaben (4191-73-5) was found to be 8.5 mg/L concentrations. Based on the value, the isopropylparaben was considered to be toxic to aquatic invertebrates but as it was readily biodegradable hence, it can be considered to be not classified as per the CLP regulations.

Thus based on the above both studies, chemical consider to be nontoxic and not classified as per the CLP classification criteria.