2,5-diaminobenzenesulphonic acid

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

For read across substance

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on two short term toxicity study of aquatic invertebrate for the test chemical :
2. 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.
3.This study was designed to assess the toxic effects of the test compound on the test Daphnids.Beaker containing 20ml of media with 10 Daphinds. A semi-static procedure was used for the study and it was conducted in compliance with the OECD guideline 202.
4.In this study, test chemical was examined for their acute aquatic toxicity employing a freshwater invertebrate (Daphnia magna).The 96 h toxicity tests with D. magna .

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

not specified

Details on test solutions:

2.The stock solution 200 mg/l was prepared by dissolving light grey powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.
3.The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg of the test substance in 500 ml of ADaM’s media. Achieving test concentrations of 100 mg/L, respectively.
4.Stock solutions were prepared with test material, from which test solutions were prepared with appropriate dilution medium for test species.

Test organisms (species):

Daphnia magna

Details on test organisms:

2: 2) 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

3: 3) TEST ORGANISM
- Common name: Water flea
- Strain/clone: No data available
- Source: Own breeding of Daphnia magna
- Age: No data available
- Feeding during test : No feeding after hatching

- Health during acclimation (any mortality observed): No data available

QUARANTINE (wild caught)
- Duration: No data available
- Health/mortality: No data available
4: 4) TEST ORGANISM
- Common name: Water Flea
- Food type: YCT (Yeast:Ceropyl®:Tetramin®) and algae (Selenastrum capricornutum)
- Frequency: daily

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Hardness:

3: 150.5 mg of CaCO3

Test temperature:

3: 20 °C ± 2 °C

pH:

2: sample at concentration 200.0 mg/l: pH = 6.9 changed to pH = 7.4 during the test,
control 1: pH = 7.6 changed to pH = 7.4 during the test,
control 2: pH = 7.7 did not change during the test,
control 3: pH = 7.7 changed to pH = 7.6 during the test
3: 7.15

Dissolved oxygen:

2: higher than 7.8 mg/L at the end of test

Nominal and measured concentrations:

2) 5.0 , 10.0 ,25.0, 50.0 , 100.0 , 200.0 mg/l
3) 100 mg/L

Details on test conditions:

2: 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

3: TEST SYSTEM
- Test vessel: The test vessels used were all glass beaker having the water capacity of 25ml.
- Material, size, headspace, fill volume: 20 ml
- Aeration: No aeration during experiment
- No. of organisms per vessel: 10 Daphnids
- No. of vessels per concentration (replicates): No data available
- No. of vessels per control (replicates): No data available
- No. of vessels per vehicle control (replicates): No data available
- Biomass loading rate: No data available

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: No data available
- Total organic carbon: No data available
- Particulate matter: No data available
- Metals: No data available
- Pesticides: No data available
- Chlorine: No data available
- Alkalinity: No data available
- Ca/mg ratio: No data available
- Conductivity: No data available
- Salinity: No data available
- Culture medium different from test medium: No data available
- Intervals of water quality measurement: No data available

OTHER TEST CONDITIONS
- Adjustment of pH: No data available
- Photoperiod: 16 hours light and 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Determination or calculation of the median effective concen trations (EC 50) and observations ((immobility, pH, Temperature, dissolved oxygen content) ) of the symptoms were done after 24 and 48 hours.

VALIDITY OF THE TEST:
1. In the control, including the control containing the solubilising agent, not more that 10 percent of the daphnids should have been immobiliz ed.
2. The dissolved oxygen concentration at the end of the test should be 3 mg/l in control and test vessels

4: TEST SYSTEM
- Test vessel: glass jars
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 6 L glass jars
- No. of organisms per vessel: 5

TEST MEDIUM / WATER PARAMETERS: Water quality parameters including hardness, alkalinity, pH, conductivity, and dissolved oxygen were routinely monitored following the standard methods by American Public Health Association, American Water Works Association, and Water Pollution Control Federation (1992).
- Intervals of water quality measurement: The culture water was renewed twice a week.

OTHER TEST CONDITIONS
- Photoperiod: Photoperiod of 16L: 8D was maintained

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobility

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: preliminary range finding tests were conducted to determine the concentration range to be used in the definitive tests. Reference tests using sodium chloride as a standard toxicant were carried out on a monthly basis to confirm the comparable sensitivity of the test organisms over time.
The acute toxicity results were reasonably predicted by a quantitative structure activity relationship model using pH-dependent distribution coefficient and molecular orbital energy parameters of the test substance.
For acute tests with the cladoceran, the median effective and lethal concentrations were calculated using ToxStat (version 3.5, West Inc.,Cheyenne, WY, USA).

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI 268.8 - 929.9 mg/l

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

158.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% confidence interval-133.0–184.4

Validity criteria fulfilled:

not specified

Conclusions:

The test chemical is not likely to be toxic to aquatic invertebrate atleast in the dose range of 100 - 500 mg/l

Executive summary:

Data available for the read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate of the test chemical .The studies are as mentioned below:

Aim of this study was to assess the short term toxicity of test material 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 200 mg/l was prepared by dissolving light grey powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.5.0 , 10.0 ,25.0, 50.0 , 100.0 , 200.0 mg/lconcentrations 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 , in Daphnia magna was determined to be 500 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be non-hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.

In another study , daphnia sp., Acute Immobilization Test according to OECD Guideline 202 was conducted for test material to assess the toxic effects of the test compound on the test Daphnids. Beaker containing 20ml of media with 10 Daphinds. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg of the test substance in 500 ml of ADaM’s media. Achieving test concentrations of 100 mg/L, respectively.

A semi-static procedure was used for the study. The nominal concentration selected for the experiment was 100 mg/L and test Daphnids were exposed to this concentration for 48hours. The Effective concentrations EC50 was found to be >100 mg/L.

Thus, according to the CLP Criteria for aquatic classification of thesubstance, it is concluded that test material does not exhibit toxicity to aquatic invertebrate (daphnia magna) andtherefore it cannot be classified as hazardous substance. The above study was further supported by data from peer reviewed journal for another read across substance,The 96 h toxicity test of test chemical with Daphnia magna were conducted in accordance with the recommended procedure outlined in peer reviewed journal.During experiment test material used as vehicle which is not used in excess of 0.5% in the final exposure solution.test condition maintained as 20 deg.C test temp. in static freshwater and Water quality parameters including hardness, alkalinity, pH, conductivity, and dissolved oxygen were routinely monitored following the standard methods by American Public Health Association, American Water Works Association, and Water Pollution Control Federation (1992).

The acute toxicity results were reasonably predicted by a quantitative structure activity relationship model using pH-dependent distribution coefficient and molecular orbital energy parameters of the test.

In experiment theEffective concentration to 50% of test organisms (EC50) on the basis of immobility effect was observed to be 174.4 mg/l with 95% confidence interval-158.4–191.9 for 48 hrs exposure period and 158.8 mg/l with 95% confidence interval-133.0–184.4 for 96 hrs exposure period for test chemical.

Based on the EC50 value, substance is likely to be non-hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.

Description of key information

Short-term toxicity to aquatic invertebrates:

Data available for the read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate of the test chemical .The studies are as mentioned below:

Aim of this study was to assess the short term toxicity of test material 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 200 mg/l was prepared by dissolving light grey powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.5.0 , 10.0 ,25.0, 50.0 , 100.0 , 200.0 mg/lconcentrations 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 , in Daphnia magna was determined to be 500 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be non-hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.

In another study , daphnia sp., Acute Immobilization Test according to OECD Guideline 202 was conducted for test material to assess the toxic effects of the test compound on the test Daphnids. Beaker containing 20ml of media with 10 Daphinds. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg of the test substance in 500 ml of ADaM’s media. Achieving test concentrations of 100 mg/L, respectively.

A semi-static procedure was used for the study. The nominal concentration selected for the experiment was 100 mg/L and test Daphnids were exposed to this concentration for 48hours. The Effective concentrations EC50 was found to be >100 mg/L.

Thus, according to the CLP Criteria for aquatic classification of thesubstance, it is concluded that test material does not exhibit toxicity to aquatic invertebrate (daphnia magna) andtherefore it cannot be classified as hazardous substance. The above study was further supported by data from peer reviewed journal for another read across substance,The 96 h toxicity test of test chemical with Daphnia magna were conducted in accordance with the recommended procedure outlined in peer reviewed journal.During experiment test material used as vehicle which is not used in excess of 0.5% in the final exposure solution.test condition maintained as 20 deg.C test temp. in static freshwater and Water quality parameters including hardness, alkalinity, pH, conductivity, and dissolved oxygen were routinely monitored following the standard methods by American Public Health Association, American Water Works Association, and Water Pollution Control Federation (1992).

The acute toxicity results were reasonably predicted by a quantitative structure activity relationship model using pH-dependent distribution coefficient and molecular orbital energy parameters of the test.

In experiment theEffective concentration to 50% of test organisms (EC50) on the basis of immobility effect was observed to be 174.4 mg/l with 95% confidence interval-158.4–191.9 for 48 hrs exposure period and 158.8 mg/l with 95% confidence interval-133.0–184.4 for 96 hrs exposure period for test chemical.

Based on the EC50 value, substance is likely to be non-hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

174.4 mg/L

Additional information

Short-term toxicity to aquatic invertebrates:

Data available for the read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate of the test chemical .The studies are as mentioned below:

Aim of this study was to assess the short term toxicity of test material 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 200 mg/l was prepared by dissolving light grey powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.5.0 , 10.0 ,25.0, 50.0 , 100.0 , 200.0 mg/lconcentrations 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 , in Daphnia magna was determined to be 500 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be non-hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.

In another study , daphnia sp., Acute Immobilization Test according to OECD Guideline 202 was conducted for test material to assess the toxic effects of the test compound on the test Daphnids. Beaker containing 20ml of media with 10 Daphinds. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg of the test substance in 500 ml of ADaM’s media. Achieving test concentrations of 100 mg/L, respectively.

A semi-static procedure was used for the study. The nominal concentration selected for the experiment was 100 mg/L and test Daphnids were exposed to this concentration for 48hours. The Effective concentrations EC50 was found to be >100 mg/L.

Thus, according to the CLP Criteria for aquatic classification of thesubstance, it is concluded that test material does not exhibit toxicity to aquatic invertebrate (daphnia magna) andtherefore it cannot be classified as hazardous substance. The above study was further supported by data from peer reviewed journal for another read across substance,The 96 h toxicity test of test chemical with Daphnia magna were conducted in accordance with the recommended procedure outlined in peer reviewed journal.During experiment test material used as vehicle which is not used in excess of 0.5% in the final exposure solution.test condition maintained as 20 deg.C test temp. in static freshwater and Water quality parameters including hardness, alkalinity, pH, conductivity, and dissolved oxygen were routinely monitored following the standard methods by American Public Health Association, American Water Works Association, and Water Pollution Control Federation (1992).

The acute toxicity results were reasonably predicted by a quantitative structure activity relationship model using pH-dependent distribution coefficient and molecular orbital energy parameters of the test.

In experiment theEffective concentration to 50% of test organisms (EC50) on the basis of immobility effect was observed to be 174.4 mg/l with 95% confidence interval-158.4–191.9 for 48 hrs exposure period and 158.8 mg/l with 95% confidence interval-133.0–184.4 for 96 hrs exposure period for test chemical.

Based on the EC50 value, substance is likely to be non-hazardous to aquatic invertebrate and cannot be classified as aquatic as per the CLP classification criteria.