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Ecotoxicological information

Endpoint summary

Administrative data

Description of key 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 partially soluble in water. The in house water solubility in DI water was 21.6523 mg/L. Therefore, the stock solution was prepared by dissolving 5 g of the test substance in 5 liters of potable water (passed through reverse osmosis system) with continuous 72 hrs stirring. After stirring the stock analytically detected and solubility in RO water found to be 122.575 mg/L. By using this value of solubility following test concentrations were prepared 6.25mg/L, 12.5mg/L, 25mg/L,50mg/L,100mg/L ,respectively and Zebra Fish Danio rerio were 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 6.25 mg/l. Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic chronic 2 as per the CLP classification criteria.

Long term toxicity to fish:

Based on the prediction done using ECOSAR class, the long term toxicity on fish was predicted for test substance. On the basis of no effects observed in a static freshwater system, the NOEC value for the substance is estimated to be 0.343 mg/l for fish for 28 days of exposure duration. Based on this value, it can be concluded that the test chemical can be considered as toxic to fish at environmentally relevant concentrations and can be considered to be classified in aquatic chronic 2 category as per the CLP classification criteria. 

 

Short term toxicity to aquatic invertebrate:

Daphnia sp., Acute Immobilization Test according to OECD Guideline 203 was conducted for test chemical. The test substance was partially soluble in water having solubility in DI water was 21.6523 mg/L. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 L of ADaM’s media. Kept it for 72 hrs stirring , after stirring stock was analytically detected, the solubility of stock in adams media found to be 316.102 mg/L.Used thses stock for further test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L, 100 mg/L respectively 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 observed to be 6.25 mg/l. Test consider to be valid because it meets the criteria as per the OECD guideline. First in the control, including the control containing the solubilising agent, not more that 10 percent of the daphnids should have been immobilized. Secondly the dissolved oxygen concentration at the end of the test should be 3 mg/l in control and test vessels. 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, does exhibit short term toxicity to Daphnia and consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

Long term toxicity to aquatic invertebrates:

Based on the prediction done using ECOSAR class, the long term toxicity on aquatic invertebrate daphnia was predicted for test substance. On the basis of no effects observed in a static freshwater system, the NOEC value for the substance is estimated to be 0.316 mg/l for daphnia for 21 days of exposure duration. Based on this value, it can be concluded that the test chemical can be considered as toxic to aquatic invertebrate daphnia at environmentally relevant concentrations and can be considered to be classified in aquatic chronic 2 category as per the CLP classification criteria. 

 

Toxicity to aquatic algae and cyanobacteria:

Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance in accordance with OECD Guideline 201 (Alga, Growth Inhibition Test). The stock solution (50 mg/L) was prepared by dissolving colourless liquid in OECD growth medium. Test solutions  of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Conc. of test chemical used for the study were 0, 0.7, 5, 1.2, 2.0, 3.5, 5.8 and 10 mg/L, respectively. The test was performed under static conditions in a static fresh water system at a temp. of 23±2°C. Initial cell density of test organism used was 5x10(3) cells/ml. Determination of cell counting involve the use of microscope with counting chamber Cyrus I or electronic particle counter.. ErC50 was calculated using non-linear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance in Desmodesmus subspicatus was determined to be 1.6 mg/L on the basis of effects on growth rate in a 72 hour study. Thus, based on this value, test chemical can be considered as toxic to aquatic organisms and thus can be classified as aquatic chronic category 2 as per the CLP criteria.

 

 

Toxicity to microorganisms:

toxicity to micro-organisms study was conducted on Photobacterium leiognathi SB strain for 30 mins. Toxscreen bioassay test was performed for the toxicity study.   All stock solutions were prepared in methanol and serially diluted in distilled water to obtain the target concentrations. The methanol concentration in the exposure solutions, including controls, was 0.01% (v/v) in distillate water in the tested solutions. 2-4-dichlorophenol (DCP) was used as control solution.   The test bacteria was exposed to the test substance with exposure duration of 15 and 30 mins. Bacterial luminescence inhibition was measured after 15 and 30 mins of exposure to tested solutions.   Based on decrease in bacterial light production by the test organism Photobacterium leiognathi SB strain, the EC50 value after 15 and 30 exposure duration with the test chemical, EC50 was observed to be 1.3 and 1.6 mg/l, respectively and LOEC value was observed to be 0.25 mg/l.

Additional information

Summarized result for the toxicity of test chemical on the growth and mortality of aquatic life’s including fish, invertebrates, algae and microorganism were studied and are as mention below:

Short term toxicity to fish:

Based on the short term toxicity studies available for the test chemical, studies were reviewed to determine the toxic nature of test chemical on the growth and mortality of fish. The studies are as mentioned below:

 

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 partially soluble in water. The in house water solubility in DI water was 21.6523 mg/L. Therefore, the stock solution was prepared by dissolving 5 g of the test substance in 5 liters of potable water (passed through reverse osmosis system) with continuous 72 hrs stirring. After stirring the stock analytically detected and solubility in RO water found to be 122.575 mg/L. By using this value of solubility following test concentrations were prepared 6.25mg/L, 12.5mg/L, 25mg/L,50mg/L,100mg/L ,respectively and Zebra Fish Danio rerio were 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 6.25 mg/l. Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic chronic 2 as per the CLP classification criteria.

 

First study was supported by the second study. 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 stock solution was prepared by dissolving 2 gm of the test substance in 2 liter of potable water (passed through reverse osmosis system) with continuous stirring of 72 hrs. After the completion of 72 hrs stirring, the stock sample was analytically detected to calculate the actual solubility and the solubility obtained was then used for achieving test concentrations of 0.625 mg/L,1.25 mg/L,2.5 mg/L,5 mg/L & 10 mg/L, respectively. 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. The nominal concentration selected for the experiment was 0.625mg/L, 1.25mg/L, 2.5mg/L, 5mg/L, 10mg/L and Zebra fish were exposed to this concentration for 96 hours. The lethal concentrations LC50 was observed to be in the range of > 2.5 mg/l to < 5 mg/l. Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test chemical on Zebra Fish Danio rerio was determine to be > 2.5 mg/l to < 5 mg/l. Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aq aquatic chronic 2 as per the CLP classification criteria.

 

Thus based on the both above studies, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

 

Long term toxicity to fish:

Based on the prediction done using ECOSAR class, the long term toxicity on fish was predicted for test substance. On the basis of no effects observed in a static freshwater system, the NOEC value for the substance is estimated to be 0.343 mg/l for fish for 28 days of exposure duration. Based on this value, it can be concluded that the test chemical can be considered as toxic to fish at environmentally relevant concentrations and can be considered to be classified in aquatic chronic 2 category as per the CLP classification criteria. 

 

Short term toxicity to aquatic invertebrate:

Various short term studies available for the test chemical were reviewed to determine the toxic nature of test chemical on the growth and mobility of aquatic invertebrates. The studies are as mentioned below:

 

In the first key study Daphnia sp., Acute Immobilization Test according to OECD Guideline 203 was conducted for test chemical. The test substance was partially soluble in water having solubility in DI water was 21.6523 mg/L. Therefore, the test solution was prepared by dissolving 1 g of the test substance in 1 L of ADaM’s media. Kept it for 72 hrs stirring , after stirring stock was analytically detected, the solubility of stock in adams media found to be 316.102 mg/L.Used thses stock for further test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L, 100 mg/L respectively 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 observed to be 6.25 mg/l. Test consider to be valid because it meets the criteria as per the OECD guideline. First in the control, including the control containing the solubilising agent, not more that 10 percent of the daphnids should have been immobilized. Secondly the dissolved oxygen concentration at the end of the test should be 3 mg/l in control and test vessels. 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, does exhibit short term toxicity to Daphnia and consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

 

Above study was supported by the second study. Determination of the inhibition of the mobility of daphnids was carried out with the test substance according to OECD Guideline 202. The stock solution 100 mg/l was prepared by dissolving colourless liquid in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the concentrations 0, 1.2, 2, 3.5, 5.8 and 10 mg/l under the static system. Effects on immobilisation were observed for 48 hours. The median effective concentration (EC50) for the test substance, in Daphnia magna was determined to be 3.6 mg/L on the basis of mobiity inhibition effects in a 48 hour study. This value indicates that the test chemical is likely to be hazardous to aquatic invertebrates and can be classified as aquatic chronic 2 category as per the CLP criteria.

Thus based on the above effects and studies available on the aquatic invertebrate, test chemical consider to be toxic to the aquatic invertebrates and hence classified in aquatic chronic 2 category as per the CLP classification criteria.

 

Long term toxicity to aquatic invertebrates:

Based on the various long term studies available for the test chemical, studies were reviewed to determine the toxic nature of test chemical on the mobility and reproduction of aquatic invertebrates. The studies are as mentioned below:

 

Based on the prediction done using ECOSAR class, the long term toxicity on aquatic invertebrate daphnia was predicted for test substance. On the basis of no effects observed in a static freshwater system, the NOEC value for the substance is estimated to be 0.316 mg/l for daphnia for 21 days of exposure duration. Based on this value, it can be concluded that the test chemical can be considered as toxic to aquatic invertebrate daphnia at environmentally relevant concentrations and can be considered to be classified in aquatic chronic 2 category as per the CLP classification criteria. 

 

Above study was supported by the second study. 21 days long term toxicity tests of test chemical were performed on daphnia magna. After the exposure of test chemical with daphnia magna for 21 days, no effects were observed on the reproduction rate of daphnia magna. Thus the NOEC was observed at 0.84 mg/l. Based on the NOEC value, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

Similar aim of this study was to access the effect of test chemical on the reproduction and growth rate of aquatic invertebrate daphnia magna. Test conducted for 10 days. Based on the low effect of test chemical on the growth and reproduction of daphnia magna after the exposure of 10 days, LOEC was observed at 0.1 mg/l. Based on the LOEC, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

 

In the fourth study long term toxicity test was performed on Daphnia magna for 21 days according to the OECD guideline 211. 20 ml test tubes were used as a vessel with different concentrations in which 1 larva at each test tubes were kept for experiment. After 48 hrs test water were replaced and count the cumulative numbers of broods. Based on that both Immobilized bodies and cumulative numbers of broods NOEC was determined. On the basis of above experiment the NOEC value for test chemical to Daphnia magna was found to be 0.84 mg/l concentration.

Thus based on the above effects and studies available on the long term effect on aquatic invertebrate, test chemical consider to be toxic to the aquatic invertebrates and hence classified in aquatic chronic 2 category as per the CLP classification criteria.

 

Toxicity to aquatic algae and cyanobacteria:

Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance in accordance with OECD Guideline 201 (Alga, Growth Inhibition Test). The stock solution (50 mg/L) was prepared by dissolving colourless liquid in OECD growth medium. Test solutions  of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Conc. of test chemical used for the study were 0, 0.7, 5, 1.2, 2.0, 3.5, 5.8 and 10 mg/L, respectively. The test was performed under static conditions in a static fresh water system at a temp. of 23±2°C. Initial cell density of test organism used was 5x10(3) cells/ml. Determination of cell counting involve the use of microscope with counting chamber Cyrus I or electronic particle counter.. ErC50 was calculated using non-linear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance in Desmodesmus subspicatus was determined to be 1.6 mg/L on the basis of effects on growth rate in a 72 hour study. Thus, based on this value, test chemical can be considered as toxic to aquatic organisms and thus can be classified as aquatic chronic category 2 as per the CLP criteria.

 

 

Toxicity to microorganisms:

Data from various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

 

In the first study toxicity to micro-organisms study was conducted on Photobacterium leiognathi SB strain for 30 mins. Toxscreen bioassay test was performed for the toxicity study.   All stock solutions were prepared in methanol and serially diluted in distilled water to obtain the target concentrations. The methanol concentration in the exposure solutions, including controls, was 0.01% (v/v) in distillate water in the tested solutions. 2-4-dichlorophenol (DCP) was used as control solution.   The test bacteria was exposed to the test substance with exposure duration of 15 and 30 mins. Bacterial luminescence inhibition was measured after 15 and 30 mins of exposure to tested solutions.   Based on decrease in bacterial light production by the test organism Photobacterium leiognathi SB strain, the EC50 value after 15 and 30 exposure duration with the test chemical, EC50 was observed to be 1.3 and 1.6 mg/l, respectively and LOEC value was observed to be 0.25 mg/l.

 

Similar All stock solutions were prepared in methanol and serially diluted in distilled water to obtain the target concentrations. The methanol concentration in the exposure solutions, including controls, was 0.01% (v/v) in distillate water in the tested solutions. Potassium dichromate (K2Cr2O7) was used as control substance. The test organism was exposed to the test substance with exposure duration 28 hrs. Inhibition in bacterial growth was measured after 24 and 28 hrs of exposure to tested solutions. Based on growth rate inhibition of test organism Tetrahymena thermophila, the EC50 value after 24 and 28 hrs exposure duration was observed to be 4.3 and 5.7 mg/l, respectively and LOEC value ewas determine to be 0.48 mg/l.

 

Thus based on both studies chemical toxicity value ranges from 0.48 to 1.6 mg/l.

Based on the above effects chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.a