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Short term toxicity to fish:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was sparingly soluble in water. Therefore, the test solution was prepared by dissolving 2 g of the test substance in 2 liters of potable water (passed through reverse osmosis system) with 72 hrs continuous stirring. After that analytically detect that stock and set up test concentrations of 3.125mg/L,6.25mg/L,12.5mg/L,25mg/L,50mg/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>12.5 mg/L but <25 mg/L.Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified aq aquatic chronic 3 as per the CLP classification criteria.

Short term toxicity to aquatic invertebrate:

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 100.0 g/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted water.10 , 20 , 30 , 45 , 100 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 43.2 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be hazardous to aquatic invertebrate can be classified as aquatic chronic 3 category as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria:

Aim of this study was to evaluate the nature of chemical test chemical when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). The stock solution 200.0 g/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and and inoculum culture. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Test concentration were used 3.8 , 4.7, 9.0 , 11.3, 90.1 mg/l. 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. Effect on the growth of algae was determine after an exposure period of 72 hrs.

The median effective concentration (EC50) for the test substance , in algae was determined to be 111.7 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Based on the EC50 value, which indicates that the substance is likely to be non-hazardous to aquatic algae and cannot be 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:

1)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.

2)In a study of toxicity of test mateial to the Marine Dinoflagellate Gymnodinium breve the effect of Potassium hydrogen phthalate 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 access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was sparingly soluble in water. Therefore, the test solution was prepared by dissolving 2 g of the test substance in 2 liters of potable water (passed through reverse osmosis system) with 72 hrs continuous stirring. After that analytically detect that stock and set up test concentrations of 3.125mg/L,6.25mg/L,12.5mg/L,25mg/L,50mg/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>12.5 mg/L but <25 mg/L.Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified aq aquatic chronic 3 as per the CLP classification criteria.

Short term toxicity to aquatic invertebrate:

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 100.0 g/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted water.10 , 20 , 30 , 45 , 100 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 43.2 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be hazardous to aquatic invertebrate can be classified as aquatic chronic 3 category as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria:

Aim of this study was to evaluate the nature of chemical test chemical when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). The stock solution 200.0 g/l was prepared by dissolving colourless liquid in acetone. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and and inoculum culture. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Test concentration were used 3.8 , 4.7, 9.0 , 11.3, 90.1 mg/l. 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. Effect on the growth of algae was determine after an exposure period of 72 hrs.

The median effective concentration (EC50) for the test substance , in algae was determined to be 111.7 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Based on the EC50 value, which indicates that the substance is likely to be non-hazardous to aquatic algae and cannot be 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:

1)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.

2)In a study of toxicity of test mateial to the Marine Dinoflagellate Gymnodinium breve the effect of Potassium hydrogen phthalate 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.