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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:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Principles of method if other than guideline:
Short term toxicity of test chemical 1,4-dichloro-2-nitrobenzene to aquatic invertebrate daphnia magna 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:
Details on test material
- Name of test material (as cited in study report): 2,5-Dichloronitrobenzene
- Molecular formula (if other than submission substance): C6-H3-Cl2-N-O2
- Molecular weight (if other than submission substance): 192.001
- Smiles notation (if other than submission substance): c1(c(ccc(c1)Cl)Cl)[N+](=O)[O-]
- InChl (if other than submission substance): 1S/C6H3Cl2NO2/c7-4-1-2-5(8)6(3-4)9(10)11/h1-3H
- Structural formula attached as image file (if other than submission substance): see Fig.
- Substance type:Organic
- Physical state:Solid
Analytical monitoring:
no
Details on test solutions:
The stock solution 20 mg/l was prepared by dissolving white powder reconstituted water. 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 20 mg/l: pH 7.9 change to 7.8
Control: pH: 8 changes to 7.9
Dissolved oxygen:
higher than 7.4 mg/L at the end of test
Nominal and measured concentrations:
0, 2.5, 4.2, 7.1, 12 and 20 mg/l were used in the study
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.:
13.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95 % CI was 10.7 to 18 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 1,4-dichloro-2-nitrobenzene in Daphnia magna was determined to be 13.9 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 1,4-dichloro-2-nitrobenzene 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 20 mg/l was prepared by dissolving white powder reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 2.5, 4.2, 7.1, 12 and 20 mg/l 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 1,4-dichloro-2-nitrobenzene in Daphnia magna was determined to be 13.9 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 and can be classified as aquatic chronic 3 category as per the CLP classification criteria.

Description of key information

Aim of this study was to assess the short term toxicity of 1,4-dichloro-2-nitrobenzene 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 20 mg/l was prepared by dissolving white powder reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 2.5, 4.2, 7.1, 12 and 20 mg/l 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 1,4-dichloro-2-nitrobenzene in Daphnia magna was determined to be 13.9 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 and can be classified as aquatic chronic 3 category as per the CLP classification criteria.

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
13.9 mg/L

Additional information

Summarized studies result of the toxicity of test chemical 1,4-dichloro-2-nitrobenzene (CAS No. 89-61-2) on the mobility of aquatic invertebrate daphnia magna were observed and mention below:

 

Aim of this study first key study was to assess the short term toxicity of 1,4-dichloro-2-nitrobenzene 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 20 mg/l was prepared by dissolving white powder reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 2.5, 4.2, 7.1, 12 and 20 mg/l 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 1,4-dichloro-2-nitrobenzene in Daphnia magna was determined to be 13.9 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 and can be classified as aquatic chronic 3 category as per the CLP classification criteria.

 

Similar study from secondary source water flea Daphnia magna was exposed to the test compound 1,4-dichloro-2- nitrobenzene during a 48 hrs study. The LC50 is determine to be 11000 µg/L (5600- 18000 µg/L) i.e. 11 mg/L (5.6- 18 mg/L). Thus based on the LC50 value, chemical considers to be toxic.

 

24 hrs short term toxicity to Daphnia magna was studied to evaluate the toxic nature of the test compound Benzene, 1, 4-Dichloro-2-Nitro. 20 daphnids (4 replicates; 5 organisms per replicate) were exposed to each of 7 nominal concentrations (1.0-32 mg/L). Stock solution was prepared with DMSO: HCO = 9:1 (1000 mg/L). Controls with and without this vehicle were taken for the test. The 24 hrs EC50 for the test compound Benzene, 1, 4-Dichloro-2-Nitro in Daphnia magna is 8.0 mg/L (95% confidence limits: 6.1-11 mg/L).

 

Thus based on the experimental key study performed according to the OECD guideline, thus on that basis chemical consider to be toxic and can be classified as aquatic chronic 3 category as per the CLP classification criteria.