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Ecotoxicological information

Endpoint summary

Administrative data

Description of key information

Short-term toxicity to fish:

Study was conducted to assess the effect of test chemical on the mortality of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 48 hrscontinuous stirring for achieving test concentrations of 0.3125mg/L,  0.625mg/L, 1.25mg/L, 2.5mg/L, 5mg/L respectively.This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L and Zebra FishDanio reriowere exposed to these concentration for 96 hours.Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to nominal test concentrations, LC50 was determine to be 0.625 mg/l . Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic acute 1 as per the CLP classification criteria.

Short-term toxicity to aquatic invertebrates:

Aim of this study was to assess the short term toxicity of test substance 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 slightly yellow powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0.5, 1, 2, 4 and 8 mg/l nominal 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 , in Daphnia magna was determined to be 2.5 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and can be be classified as aquatic chronic 2 category as per the CLP classification criteria.

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 in each of these test vessels was kept constant which is 60 ml so that a sufficient amount of head space was left. The test substance was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200mg/L. This stock solution was kept for stirring/ sonication for 0 minutes to obtain a homogenous solution for the experiment. The test concentrations0.5 mg/L,1 mg/L,2 mg/L,4 mg/L,8 mg/L,16 mg/Lwere 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.

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 10.580 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was hazrdous and can be consider to be classified as aquatic chronic 2 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:

Photobacterium phosphoreum was test organism used to determined the toxicity of test material done by Microtox test method . The time period was 5 to 30 minute at 15 degree C and pH range from 5 to 9. After exposure, the 50% Effective concentration value for 1 test chemical to Photobacterium phosphoreum was determined to be 3.02 mg/l based on viability and sensitivity of bacteria.

In another study for structurally similar read across substance , Pseudomonas respiratory arrest test was carried out for the test material substance.The 50% Effective concentration (EC50) of the substance5is determined to be >10000 mg/L based on inhibitory effect on Oxygen consumption (respiration).

Additional information

Short-term toxicity to fish:

Study was conducted to assess the effect of test chemical on the mortality of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 48 hrscontinuous stirring for achieving test concentrations of 0.3125mg/L,  0.625mg/L, 1.25mg/L, 2.5mg/L, 5mg/L respectively.This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L and Zebra FishDanio reriowere exposed to these concentration for 96 hours.Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to nominal test concentrations, LC50 was determine to be 0.625 mg/l . Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic acute 1 as per the CLP classification criteria.

Short-term toxicity to aquatic invertebrates:

Short-term toxicity to aquatic invertebrates was evaluated based on two experimental study report,

Aim of this study was to assess the short term toxicity of test substance 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 slightly yellow powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 0.5, 1, 2, 4 and 8 mg/l nominal 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 , in Daphnia magna was determined to be 2.5 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and can be be classified as aquatic chronic 2 category as per the CLP classification criteria.

In another study report ,daphnia sp., Acute Immobilization Test according to OECD Guideline 203 was conducted for test material. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 500 mg of the test substance in 500 ml of ADaM’s media. Achieving test concentrations of 1 g/L, respectively.

The nominal concentration selected for the experiment were 0.625 mg/L,1.25 mg/L, 2.5 mg/L, 5 mg/L, and 10 mg/L and test Daphnia magna were exposed to these concentration for 48 hours. The median lethal concentration (EC50) for test chemical on Daphnia magna in a 48 hours study on the basis of immobilization effect was found to be 0.625 mg/l.Thus, on the basis of this EC50 value and according to CLP criteria for aquatic classification of the substance, it is concluded that the substance exhibits short term toxicity to Daphnia.

After 48 hours of exposure to test item to nominal test concentrations, EC50 was determine to be 0.62mg/l . Based on the EC50, it can be consider that the chemical was and can be consider to be classified as aquatic acute 1 as per the CLP classification criteria.

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 in each of these test vessels was kept constant which is 60 ml so that a sufficient amount of head space was left. The test substance was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200mg/L. This stock solution was kept for stirring/ sonication for 0 minutes to obtain a homogenous solution for the experiment. The test concentrations0.5 mg/L,1 mg/L,2 mg/L,4 mg/L,8 mg/L,16 mg/Lwere 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.

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 10.580 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was hazrdous and can be consider to be classified as aquatic chronic 2 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:

Photobacterium phosphoreum was test organism used to determined the toxicity of test material done by Microtox test method . The time period was 5 to 30 minute at 15 degree C and pH range from 5 to 9. After exposure, the 50% Effective concentration value for 1 test chemical to Photobacterium phosphoreum was determined to be 3.02 mg/l based on viability and sensitivity of bacteria.

In another study for structurally similar read across substance , Pseudomonas respiratory arrest test was carried out for the test material substance.The 50% Effective concentration (EC50) of the substance5is determined to be >10000 mg/L based on inhibitory effect on Oxygen consumption (respiration).