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Ecotoxicological information

Endpoint summary

Administrative data

Description of key information

Short term toxicity to fish:

Fish Acute Toxicity 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 62.5 mg, 125 mg, 250 mg, 500 mg& 1 g of the test substance in 10 liters of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/l & 100 mg/L, respectively.The nominal concentration selected for the experiment were 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/l & 100 mg/L and Zebra fish were exposed to these concentration for 96 hours. The fishes were slowly swimming without showing any abnormal symptoms. No mortalities were found in the control aquaria.The lethal concentrations LC0 was found to be 50 mg/L. And LC50 was calculated to be 75 mg/l.

Long term toxicity to fish:

The long term toxicity on fish was predicted for test substance by using EPI suite ECOSAR version 1.1. On the basis of effects observed in a static freshwater system, the NOEC value for the test substance was estimated to be 14.247 mg/l for fish for 28 days of exposure duration. By considering the NOEC value, it can be concluded that the test chemical can not be considered as toxic to fish at environmentally relevant concentrations and cannot be classified aquatic as per the CLP classification criteria.

Short term toxicity to aquatic invertebrate:

The Lethal concentration (LC50) value of test material in aquatic invertebrate (Daphnia Magna) in a 48 hr study based on mortality effect was estimated to be 82.747 mg/L. Thus considering the value from CLP Criteria for aquatic classification of the substance , it is concluded that test material exhibit short term toxicity aquatic invertebrate (Daphnia Magna) and can be classified aquatic chronic 3 .

Long term toxicity to aquatic invertebrate:

The long term toxicity on aquatic invertebrate was predicted for test substance by using ECOSAR version 1.1. On the basis of effects observed in a static freshwater system, the NOEC value for the substance was estimated to be 8.086 mg/l for aquatic invertebrate for 21 days of exposure duration. By considering NOEC value, it is concluded that the test chemical can be considered as not toxic to aquatic invertebrate and can not be classified 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 item  was prepared by adding 25mg of test item in 250 ml of BBM to get the final concentration of 100mg/L. This stock solution was kept for sonication for 45 minutes to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment.

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 53.190 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was toxic and can be consider to be not classified as per the CLP classification criteria.

Toxicity to microorganism:

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

Toxicity of test chemical were studied based on the effect observed on microorganisms. The inhibitory growth concentration (IGC50) value of test chemical in microorganism (Tetrahymena pyriformis) in a 48 hrs study on the basis of growth inhibition effect was determined to be 17 mg/L.

In another study of functionally similar read across substance study was conducted to evaluate the effect of test chemical on the Proliferation ratio of Tetrahymena pyriformis (Ciliate). Test conducted under the static system for 60 hrs. Based on the inhibition of proliferation rate of test organism Tetrahymena pyriformis (Ciliate) by the chemical exposure for 60 hrs, the EC50 was determine to be 66.34 mg/l.The effect concentration of test material on Tetrahymena pyriformis (Ciliate) was observed to be in the range of 17-66.34 mg/l

Additional information

Short term toxicity to fish:

Fish Acute Toxicity 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 62.5 mg, 125 mg, 250 mg, 500 mg& 1 g of the test substance in 10 liters of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/l & 100 mg/L, respectively.The nominal concentration selected for the experiment were 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/l & 100 mg/L and Zebra fish were exposed to these concentration for 96 hours. The fishes were slowly swimming without showing any abnormal symptoms. No mortalities were found in the control aquaria.The lethal concentrations LC0 was found to be 50 mg/L. And LC50 was calculated to be 75 mg/l.

Long term toxicity to fish:

The long term toxicity on fish was predicted for test substance by using EPI suite ECOSAR version 1.1. On the basis of effects observed in a static freshwater system, the NOEC value for the test substance was estimated to be 14.247 mg/l for fish for 28 days of exposure duration. By considering the NOEC value, it can be concluded that the test chemical can not be considered as toxic to fish at environmentally relevant concentrations and cannot be classified aquatic as per the CLP classification criteria.

Short term toxicity to aquatic invertebrate:

The short term toxicity of test material on aquatic invertebrate was evaluated using prediction along with data for functionally and structurally similar read across substance:

The Lethal concentration (LC50) value of test material in aquatic invertebrate (Daphnia Magna) in a 48 hr study based on mortality effect was estimated to be 82.747 mg/L. Thus considering the value from CLP Criteria for aquatic classification of the substance , it is concluded that test material exhibit short term toxicity aquatic invertebrate (Daphnia Magna) and can be classified aquatic chronic 3 .

The above prediction was supported by data for structurally similar read across substance,determination of the inhibition of the mobility of daphnids was carried out with the test substance according to OECD Guideline 202. The stock solution (200 g/L) was prepared by dissolving white powder in DMSO. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the concentrations 0, 5, 10, 20, 30, 40, 80 mg/L and the immobilisation effects were observed for 48 hours. 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 26 mg/L on the basis of mobiity inhibition effects in a 48 hour study. This value indicates that the substance is likely to be hazardous to aquatic invertebrates can can be classified as Aquatic chronic category 3 as per the CLP criteria.

The above study was further supported by data for functionally similar read accross substance, experimental study for inhibition of the mobility of daphnids was carried out with the benzimidazole according to OECD Guideline 202. The animals used for the test shall be less than 24 h old and should not be of first brood progeny. Test solutions of required concentration were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the concentrations 0, 7.5, 15, 30, 60 and 120 mg/L. The test was performed under static conditions in a static fresh water system at 20±1°C temperature. Effects on immobilisation were observed for 48 hours. EC50 was calculated using non linear regression by the software Prism 4.0. After the experiment, the median effective concentration (EC50) for the test substance, benzimidazole, in Daphnia magna was determined to be 55.5 mg/L on the basis of immobilisation effects. Based on the EC50 value chemical consider to be toxic and classified as aquatic chronic 3 as per the CLP classification criteria.

Long term toxicity to aquatic invertebrate:

The long term toxicity on aquatic invertebrate was predicted for test substance by using ECOSAR version 1.1. On the basis of effects observed in a static freshwater system, the NOEC value for the substance was estimated to be 8.086 mg/l for aquatic invertebrate for 21 days of exposure duration. By considering NOEC value, it is concluded that the test chemical can be considered as not toxic to aquatic invertebrate and can not be classified 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 item  was prepared by adding 25mg of test item in 250 ml of BBM to get the final concentration of 100mg/L. This stock solution was kept for sonication for 45 minutes to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment.

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 53.190 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was toxic and can be consider to be not classified as per the CLP classification criteria.

Toxicity to microorganism:

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

Toxicity of test chemical were studied based on the effect observed on microorganisms. The inhibitory growth concentration (IGC50) value of test chemical in microorganism (Tetrahymena pyriformis) in a 48 hrs study on the basis of growth inhibition effect was determined to be 17 mg/L.

In another study of functionally similar read across substance study was conducted to evaluate the effect of test chemical on the Proliferation ratio of Tetrahymena pyriformis (Ciliate). Test conducted under the static system for 60 hrs. Based on the inhibition of proliferation rate of test organism Tetrahymena pyriformis (Ciliate) by the chemical exposure for 60 hrs, the EC50 was determine to be 66.34 mg/l.The effect concentration of test material on Tetrahymena pyriformis (Ciliate) was observed to be in the range of 17-66.34 mg/l