6-aminonaphthalene-2-sulphonic acid

Endpoint summary

Description of key information

Short-term toxicity to fish:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for test substance to assess the toxic effects of the test compound on the Zebra fish(Danio rerio). Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study. The nominal concentration selected for the experiment were 100 mg/Land test fish were exposed to these concentration for 96 hours. The median lethal concentrations LC50 for test material on Danio rerio in a 96 hours study on the basis of mortality effect was found to be > 100 mg/L. Thus, on the basis of this LC50 value and according to CLP criteria for aquatic classification of the substa nce, it is concluded that the test substance, does not exhibit short term toxicity to fish. LC0 (96 hours) (highest loading at which no mortality was observed) =100 mg/L . Hence , based on the above effect concentrationit can be concluded that test material has no effect on aquatic invertebrates and can not be classified as per CLP criteria.

Short-term toxicity to aquatic invertebrates:

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.

Toxicity to aquatic algae and cyanobacteria:

The study was designed to assess the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test).

Test was carried out in 100mL conical flasks which were carefully autoclaved and sterilized.The test solution was prepared in aseptic condition. The test item  was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring for 24 hours to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit.The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial of the culture was kept 1 X 10E4cells/ml.

For the assessment of algal growth, the test was conducted in replicates. The control flask was maintained in triplicates as recommended in the OECD guideline and the test concentration were selected in geometric series which were maintained in duplicates. To obtain a quantitative concentration-response relationship by regression analysis, a linearizing transformation of the response data into probit was performed. Using the same, effective concentration (EC) were determined.

Algal growth was calculated daily by counting the cells microscopically with the help of haemocytometer. For microscopic observations the cultures were observed daily with the help of a microscope to verify a normal and healthy appearance of the algal culture and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test item). Apart from this, the cell count of each test vessel was also noted with the help of a microscope and haemocytometer. By spectrophotometer the absorbance values of each test vessel and control vessel was noted at 680nm.The BBM was taken as blank for both control and test vessels. The absorbance value of each vessel was in line with the average specific growth rate.

As per OECD 201, the biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72 hr test period. This corresponds to a specific growth rate of 0.92 per day. Thus, the observed specific growth rate in the control cultures during the experiment was 0.358 per day. Secondly the mean coefficient of variation for section by section specific growth rates (days 0-1, 1-2 & 2-3, for 72 hr tests) in the control cultures must not exceed 35%. Thus, the observed mean coefficient of variation in the control cultures during the experiment was 33.42%. Thirdly the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 10%. Thus, the observed coefficient of variation of average specific growth rates during the experiment in control cultures was 8.26%. Hence, the test is considered valid as per OECD guideline, 201

After 72 hours of exposure to test item to various nominal test concentrations, EC50 was determine to be >200 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria.

Toxicity to microorganisms:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the toxicity of microorganism of the test chemical .The studies are as mentioned below:

This study was examined to assess the toxic effects of the test compound on Pseudomonas fluorescens in a 24 hours of exposure. The effective concentration (EC0) value of test material in bacteria in a 24 hours of exposure on the basis of mortality effect was observed to be in the range 500- 1000 mg/L.

Additional information

Short-term toxicity to fish:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for test substance to assess the toxic effects of the test compound on the Zebra fish(Danio rerio). Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study. The nominal concentration selected for the experiment were 100 mg/Land test fish were exposed to these concentration for 96 hours. The median lethal concentrations LC50 for test material on Danio rerio in a 96 hours study on the basis of mortality effect was found to be > 100 mg/L. Thus, on the basis of this LC50 value and according to CLP criteria for aquatic classification of the substa nce, it is concluded that the test substance, does not exhibit short term toxicity to fish. LC0 (96 hours) (highest loading at which no mortality was observed) =100 mg/L . Hence , based on the above effect concentrationit can be concluded that test material has no effect on aquatic invertebrates and can not be classified as per CLP criteria.

Short-term toxicity to aquatic invertebrates:

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.

Toxicity to aquatic algae and cyanobacteria:

The study was designed to assess the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test).

Test was carried out in 100mL conical flasks which were carefully autoclaved and sterilized.The test solution was prepared in aseptic condition. The test item  was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring for 24 hours to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit.The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial of the culture was kept 1 X 10E4cells/ml.

For the assessment of algal growth, the test was conducted in replicates. The control flask was maintained in triplicates as recommended in the OECD guideline and the test concentration were selected in geometric series which were maintained in duplicates. To obtain a quantitative concentration-response relationship by regression analysis, a linearizing transformation of the response data into probit was performed. Using the same, effective concentration (EC) were determined.

Algal growth was calculated daily by counting the cells microscopically with the help of haemocytometer. For microscopic observations the cultures were observed daily with the help of a microscope to verify a normal and healthy appearance of the algal culture and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test item). Apart from this, the cell count of each test vessel was also noted with the help of a microscope and haemocytometer. By spectrophotometer the absorbance values of each test vessel and control vessel was noted at 680nm.The BBM was taken as blank for both control and test vessels. The absorbance value of each vessel was in line with the average specific growth rate.

As per OECD 201, the biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72 hr test period. This corresponds to a specific growth rate of 0.92 per day. Thus, the observed specific growth rate in the control cultures during the experiment was 0.358 per day. Secondly the mean coefficient of variation for section by section specific growth rates (days 0-1, 1-2 & 2-3, for 72 hr tests) in the control cultures must not exceed 35%. Thus, the observed mean coefficient of variation in the control cultures during the experiment was 33.42%. Thirdly the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 10%. Thus, the observed coefficient of variation of average specific growth rates during the experiment in control cultures was 8.26%. Hence, the test is considered valid as per OECD guideline, 201

After 72 hours of exposure to test item to various nominal test concentrations, EC50 was determine to be >200 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria.

Toxicity to microorganisms:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the toxicity of microorganism of the test chemical .The studies are as mentioned below:

This study was examined to assess the toxic effects of the test compound on Pseudomonas fluorescens in a 24 hours of exposure. The effective concentration (EC0) value of test material in bacteria in a 24 hours of exposure on the basis of mortality effect was observed to be in the range 500- 1000 mg/L.