Registration Dossier

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 25 µl, 50 µl, 100 µl , 200 µl and 400 µl   of the test substance in 4 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, 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 various nominal test concentrations, LC50 was determine to be> 50 mg/L<100mg/l . Based on the LC50, it can be consider that the chemical was and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.

Short-term toxicity to aquatic invertebrates:

Data available for the structurally and functionally similar read across substance has been reviewed to determine the short term toxicity of aquatic invertebrate .The studies are as mentioned below:

Aim of this study was to assess the short term toxicity of 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 colourless liquid of purity 99.62% 100  mg/l was prepaired by dissolving the substance into reconstitued test water.100 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 inhibition perccentage [%] for the test substance , in Daphnia magna was determined to be 4.0% on the basis of mobility inhibition effects in a 48 hour study.24 out of 25 daphnids were observed to be mobile in the sample. Based on the [I%] value, indicates that the substance is likely to be non-hazardous to aquatic invertebrates and cannot as per the CLP criteria.

The above eperimental data was further supported by data summaried in authoritative database further explaining the toxicity of structurally similar read across substance for its potential to cause toxicity to aquatic invertebrate.

Short term toxicity to aquatic invertebrate was evaluated for 48 h according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The EC50 value after 48 h of test material exposure was observed to be 230 mg/l.

Based on the above effect concentration (EC50) ,it can be considered that test substance is not toxic for fish and can not be classified as per CLP criteria.

Experimental data available for functionally similar read across substance was used to further support the toxicity of aquatic invertebrate,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 10mg/l was prepared by dissolving white powder in reconstituted water. The test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 50, 70,100,140 and 200 mg/L 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 189.2 mg/L on the basis of mobiity inhibition effects in a 48 hour study.

Based on the median effective concentration value, it is concluded that the substance is likely to be non-hazardous to aquatic invertebrates and cannot be classified as per the CLP criteria.

Toxicity to aquatic algae and cyanobacteria:

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

For a read across substance ,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).

The test substance test material was prepared by adding 75 mg of test substance in 150 ml of BBM to get the final concentration of 500 mg/L. The sock solution was ultrasonically agitated for 30 minutes to obtain a homogenous solution for the experiment. The remaining test solutions were prepared by dilution from the above prepared stock solution under aseptic conditions. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104 cells/ml.

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 concentrations6.25 mg/l, 12.5 mg/l, 25 mg/l, 50 mg/l, 100 mg/l and 200mg/l 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.

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

For another read across , 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 substance , was prepared by adding 106.25 mg of test substance in 250 ml of BBM to get the final concentration of 425 mg/L. The sock solution was ultrasonically agitated for 30 minutes to obtain a homogenous solution for the experiment. The remaining test solutions were prepared by dilution from the above stock solution, under aseptic condition. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104 cells/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 138.96 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was non-toxic and can be consider to be not classified as per the CLP classification criteria.

Data from read across substance was used to further support the above data , 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 concentrations 6.25 mg/l, 12.5 mg/l, 25 mg/l, 50 mg/l, 100 mg/l and 200mg/l 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. 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 to various nominal test concentrations, EC50 was determine to be >200 mg/l graphically and through probit analysis.

Based on the EC50 values , 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 microorganism:

Data available for the test chemicals has been reviewed to determine the toxicity of microorganism of the test chemical .The studies are as mentioned below:

2)Test material was used as a test material to evaluate toxicity to microorganisms as per OECD guideline 209. Activated sludge was used as the test culture at pH 7.5 for 3 h The EC50 value on the basis of respiration inhibition test was observed to be >1000 mg/l.

3)In a study of toxicity of test mateial to the Marine Dinoflagellate Gymnodinium breve the effect of test material was evaluated. The test substance was tested in a concentration of 0,1,10, 20, 50, 100, 200, 500, 1000 and 1000 ppm. The results show Median growth when concentration of test material (ppm) versus the growth rate (linear axis of graph), expressed as a percentage of the growth of a "no-add" control at 498.52 and 997 mg/l. Therefore, EC50 of test material was considered to be be 498.52 and 997 mg/l when tested on Marine Dinoflagellate Gymnodinium breve for 96 hours.

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 25 µl, 50 µl, 100 µl , 200 µl and 400 µl   of the test substance in 4 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, 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 various nominal test concentrations, LC50 was determine to be> 50 mg/L<100mg/l . Based on the LC50, it can be consider that the chemical was and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.

Short-term toxicity to aquatic invertebrates:

Data available for the structurally and functionally similar read across substance has been reviewed to determine the short term toxicity of aquatic invertebrate .The studies are as mentioned below:

Aim of this study was to assess the short term toxicity of 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 colourless liquid of purity 99.62% 100  mg/l was prepaired by dissolving the substance into reconstitued test water.100 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 inhibition perccentage [%] for the test substance , in Daphnia magna was determined to be 4.0% on the basis of mobility inhibition effects in a 48 hour study.24 out of 25 daphnids were observed to be mobile in the sample. Based on the [I%] value, indicates that the substance is likely to be non-hazardous to aquatic invertebrates and cannot as per the CLP criteria.

The above eperimental data was further supported by data summaried in authoritative database further explaining the toxicity of structurally similar read across substance for its potential to cause toxicity to aquatic invertebrate.

Short term toxicity to aquatic invertebrate was evaluated for 48 h according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The EC50 value after 48 h of test material exposure was observed to be 230 mg/l.

Based on the above effect concentration (EC50) ,it can be considered that test substance is not toxic for fish and can not be classified as per CLP criteria.

Experimental data available for functionally similar read across substance was used to further support the toxicity of aquatic invertebrate,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 10mg/l was prepared by dissolving white powder in reconstituted water. The test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. 0, 50, 70,100,140 and 200 mg/L 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 189.2 mg/L on the basis of mobiity inhibition effects in a 48 hour study.

Based on the median effective concentration value, it is concluded that the substance is likely to be non-hazardous to aquatic invertebrates and cannot be classified as per the CLP criteria.

Toxicity to aquatic algae and cyanobacteria:

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

For a read across substance ,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).

The test substance test material was prepared by adding 75 mg of test substance in 150 ml of BBM to get the final concentration of 500 mg/L. The sock solution was ultrasonically agitated for 30 minutes to obtain a homogenous solution for the experiment. The remaining test solutions were prepared by dilution from the above prepared stock solution under aseptic conditions. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104 cells/ml.

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 concentrations6.25 mg/l, 12.5 mg/l, 25 mg/l, 50 mg/l, 100 mg/l and 200mg/l 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.

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

For another read across , 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 substance , was prepared by adding 106.25 mg of test substance in 250 ml of BBM to get the final concentration of 425 mg/L. The sock solution was ultrasonically agitated for 30 minutes to obtain a homogenous solution for the experiment. The remaining test solutions were prepared by dilution from the above stock solution, under aseptic condition. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104 cells/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 138.96 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was non-toxic and can be consider to be not classified as per the CLP classification criteria.

Data from read across substance was used to further support the above data , 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 concentrations 6.25 mg/l, 12.5 mg/l, 25 mg/l, 50 mg/l, 100 mg/l and 200mg/l 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. 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 to various nominal test concentrations, EC50 was determine to be >200 mg/l graphically and through probit analysis.

Based on the EC50 values , 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 microorganism:

Data available for the test chemicals has been reviewed to determine the toxicity of microorganism of the test chemical .The studies are as mentioned below:

2)Test material was used as a test material to evaluate toxicity to microorganisms as per OECD guideline 209. Activated sludge was used as the test culture at pH 7.5 for 3 h The EC50 value on the basis of respiration inhibition test was observed to be >1000 mg/l.

3)In a study of toxicity of test mateial to the Marine Dinoflagellate Gymnodinium breve the effect of test material was evaluated. The test substance was tested in a concentration of 0,1,10, 20, 50, 100, 200, 500, 1000 and 1000 ppm. The results show Median growth when concentration of test material (ppm) versus the growth rate (linear axis of graph), expressed as a percentage of the growth of a "no-add" control at 498.52 and 997 mg/l. Therefore, EC50 of test material was considered to be be 498.52 and 997 mg/l when tested on Marine Dinoflagellate Gymnodinium breve for 96 hours.