Sodium 3-nitrobenzenesulphonate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22.06.2016 - 24.06.2016

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 in a static system.

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

yes

Details on test solutions:

The solution of white pearl 100 mg/l was prepared by dissolving the substance 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
- 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:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

± 1 hr

Test temperature:

20±1°C

pH:

Test: 8.2 (changed to 8 during test)
Control: 7.9 (changed to 8 during test)

Dissolved oxygen:

higher than 7.9 mg/L at the end of test

Nominal and measured concentrations:

Limit test conducted at 100 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:

48 h

Dose descriptor:

other: IC8

Remarks:

%

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Only 8% inhibition was observed at 100 mg/l.

Details on results:

As only 8 % inhibition were observed at 100 mg/l, thus it was concluded that the EC50 was > 100 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:

IC was calculated using non linear regression by the software Prism 4.0

Validity criteria fulfilled:

yes

Conclusions:

Based on the immobility of daphnia magna due to the exposure of test chemical for 48 hrs, the inhibitory concentration IC were observed at 100 mg/l. At 100 mg/l only 8 % inhibition was observed in daphnia magna.

Executive summary:

Aim of this study was to assess the short term toxicity of test chemical 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 solution of white pearl 100 mg/l was prepared by dissolving the substance in reconstituted water. Test conducted at the limit concentration 100 mg/l. 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. Based on the immobility of daphnia magna due to the exposure of chemical for 48 hrs, the inhibitory concentration IC were observed at 100 mg/l. At 100 mg/l only 8 % inhibition was observed in daphnia magna thus it was concluded that the EC50 was > 100 mg/l. Based on the above effects it was concluded that the chemical was nontoxic and can be consider to be not classified as per the CLP classification criteria. 

Description of key information

Aim of this study was to assess the short term toxicity of test chemical 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 solution of white pearl 100 mg/l was prepared by dissolving the substance in reconstituted water. Test conducted at the limit concentration 100 mg/l. 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. Based on the immobility of daphnia magna due to the exposure of chemical for 48 hrs, the inhibitory concentration IC were observed at 100 mg/l. At 100 mg/l only 8 % inhibition was observed in daphnia magna thus it was concluded that the EC50 was > 100 mg/l. Based on the above effects it was concluded that the chemical was nontoxic and can be consider to be not classified as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

Summarized result for the toxicity of chemical on the growth of aquatic invertebrates are as follows:

 

In the first key study from toxicity were measured. Aim of this study was to assess the short term toxicity of test chemical 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 solution of white pearl 100 mg/l was prepared by dissolving the substance in reconstituted water. Test conducted at the limit concentration 100 mg/l. 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. Based on the immobility of daphnia magna due to the exposure of chemical for 48 hrs, the inhibitory concentration IC were observed at 100 mg/l. At 100 mg/l only 8 % inhibition was observed in daphnia magna thus it was concluded that the EC50 was > 100 mg/l. Based on the above effects it was concluded that the chemical was nontoxic and can be consider to be not classified as per the CLP classification criteria. 

 

Similarly in the second supporting study for the test chemical from peer reviewed journal measured the toxicity. Aim of this study was to determine the nature and behavior of chemical when comes in contact with the test organism daphnia magna. Study conducted under static system for the total exposure of 96 hrs. Water flea collected from Put-In-Bay, Ohio initially and then cultivate and maintain in the laboratory. Effect were observed after 24, 48, 72 and 96 hrs in the form of TLm (Median tolerance limits). Based on the mortality of daphnia magna by the chemical exposure for 96 hrs, the LC50 was determine to be 5067 mg/l. And after the 24, 48hrs and 72hrs, the LC50 was observed at 8665 mg/l and 6017 mg/l. based on the LC50, chemical was consider as nontoxic and not classified as per the CLP classification criteria.

 

In the third supporting study the effective concentration EC0 and EC50 value of sodium 3-nitrobenzenesulphonate in aquatic invertebrate (Daphnia magna) in a 100 hr study on mortality effect was found to be 1000 mg/L and 2335 mg/L respectively. Thus, considering the CLP Criteria for aquatic classification of the substance, it is concluded that the test chemical does not exhibit short term toxicity to aquatic invertebrate (Daphnia magna).

Based on the experimental studies from various sources, it was concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.