Registration Dossier

Ecotoxicological information

Endpoint summary

Administrative data

Description of key information

Short term toxicity to fish

1: Based on the mortality of freshwater fish Brachydanio rerio by the chemical exposure for 96 hrs, the LC50 was determine to be 17.6 mg/l and the LC0 and LC100 was 11.3 mg/l and 26.2 mg/l.

2: Based on the mortality of fishes Pimephales promelas after the exposure of chemical for 96 hrs, the LC50 was determine to be 18.9 mg/l.

Thus based on the above both studies, chemical consider to be toxic and classified as aquatic chronic 3 as per the CLP classification criteria.

 

Long term toxicity to fish

Using the EPI Suite ECOSAR, the long term toxicity on fish was predicted for test substance. On the basis of no effects observed in a flow-through system, the no observed effect concentration (NOEC) value for the substance is estimated to be 0.164 mg/l for saltwater fish for 28 d duration. Thus on the basis of NOEC value, chemical consider to be toxic and classified in aquatic chronic category 2 as per the CLP classification criteria.

 

Short term toxicity to aquatic invertebrates

An acute immobilisation test was used to test how a range of concentrations of test chemical exerts different degrees of toxic effects on the swimming capability of Daphnia magna under otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 202. The testing aim was to determine a EC50 after 48 hours of exposure to D. magna. The stock solution used for the exposure assessment was prepared by dissolving colourless dense liquid in acetone. Daphnids were exposed to chemical in 50 ml beakers in a volume of 25 ml of liquid solution containing both the chemical and media as specified in OECD 202. The beakers were placed in a temperature controlled room at 20±1 degrees Celsius. The D. magna (age ≤24) used for the test. The animals were exposed to stock solution prepared by adding dense colourless liiquid in acetone. There were 5 Daphnia per test vessels and 4 replicates per concentration. The positive control/reference substance used in the teste showed an expected result and gave a EC50 that corresponded to previous exposures with this chemical in D. magna. Immobilisation effects in D. mangaby chemical exposure was evident after 48 hours in exposure concentrations above 1 mg/L (i.e. EC20). The EC50 was defined as a concentration that immobilizes 50% of the exposed D. magna. The % of immobilization in D. magna after 48 hours of exposure to test chemical were used in a nonlinear regression by Graphpad Software Prism 4.0 (San Diego, US). Based on the immobility of daphnia magna by the chemical exposure for 48 hrs, the EC50 was determine to be 2.59 mg/l (95% C.I: 1.81-3.7 mg/L). 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 classification criteria.

 

Long term toxicity to aquatic invertebrates

The test substance was used to evaluate its adverse effects to Daphnia magna. The test was conducted in accordance with the OECD 211 guidleines, for the period of 21 days. The test solutions was prepared in the ADAMS medium, the test concentration chosen for the exposure was 0.25 - 4 mg/L, with the geometric series of 2. the test was performed in semi static conditions, wherein the test chemical was renewed every alternate day, of the exposure period. The EC10 was calculated to be 0.148±0.85 mg/L. Based on the Effective concentration it can be classified into chronic 2 category of CLP criteria.

 

Toxicity to aquatic algae and cyanobacteria

Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance according to the OECD Guideline 201. The stock solution (32 g/L) was prepared by dissolving colourless dense liquid in acetone. 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, 2, 4, 8.5, 16, 32 and 68 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 (ErC50) for the test substance in Desmodesmus subspicatus was determined to be 47.0 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 3 as per the CLP classification criteria.

 

Toxicity to micro-organisms

The test chemical is likely to be toxic to microorganism  atleast in the concentartion range of 1 to 8 mg/l.

Additional information

Short term toxicity to fish:

Data available for the test chemical, structually and functionally similar read across chemicals has been reviewed to determine its effect on the mortality of fishes. The studies are as mentioned below:

 

Aim of this study was to determine the nature of chemical on the mortality of freshwater fish Brachydanio rerio for total exposure of 96 hrs. Chemical was analytically monetarized by GC. Test conducted under static system. After incubation period of 96 hrs, slow and inactive swimming behaviour and loss of equilibrium (uncontrolled movements) were also observed. Based on the mortality of freshwater fish Brachydanio rerio by the chemical exposure for 96 hrs, the LC50 was determine to be 17.6 mg/l and the LC0 and LC100 was 11.3 mg/l and 26.2 mg/l. Based on the LC50, chemical was consider as toxic and can be consider to to be classfied as aquatic chronic 3 category as per the CLP classification criteria.

 

Similar study was also conducted on other fish. Aim of this study was to determine the nature of chemical on the mortality of fish Pimephales promelas for total exposure of 96 hrs. Study was conducted comparable to standard method and in accordance with general accepted scientific standards. Nominal 5 concentrations in the range of 4.39 to 24.6 mg/l were used and the sample was analytically monetarized by GC method. 0.079 g 30 days old Pimephales promelas (Fathead Minnow) were used. Test conducted under the flow- through system. Observations of fish behaviour and body morphology at regular intervals. Affected fish lost schooling behavior, were hypoactive and underreactive to external stimuli. They had increased respiration were darkly colored and lost equilibrium prior to death. Based on the mortality of fishes Pimephales promelas after the exposure of chemical for 96 hrs, the LC50 was determine to be 18.9 mg/l. based on the LC50, chemical was consider as toxic and can be consider to not classified as aquatic chronic 3 category as per the CLP classification criteria.

Thus based on the above both studies, chemical consider to be toxic and classified as aquatic chronic 3 as per the CLP classification criteria.

Long term toxicity to fish:

Various experimental and predicted studies available for the test chemical and functionally similar read across chemical were reviewed to determine the toxic nature of test chemical on the fish. The studies are as mentioned below:

 

Using the EPI Suite ECOSAR, the long term toxicity on fish was predicted for test substance. On the basis of no effects observed in a flow-through system, the no observed effect concentration (NOEC) value for the substance is estimated to be 0.164 mg/l for saltwater fish for 28 d duration. Thus on the basis of NOEC value, chemical consider to be toxic and classified in aquatic chronic category 2 as per the CLP classification criteria.

 

Above data was supported by the second study from authoritative database. Aim of this study was to determine the prolonged effect of test chemical on the freshwater fish. Test conducted for 14 days under semi-static system. After the exposure of test chemical with fish for 14 days, LC50 was determine at 0.78 mg/l and no effect (NOEC) were observed at 0.14 mg/l. Thus based on the LC50 value, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

 

Objective of this third study was to determine the prolonged effect of test chemical on the fish by providing the exposure period of 14 days. After the exposure of chemical with fish for 14 days, no effects were observed on fish at concentration 0.75 mg/l. Thus on the basis of NOEC value, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

 

Thus by considering the above effects, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

 

Short term toxicity to aquatic invertebrate:

An acute immobilisation test was used to test how a range of concentrations of test chemical exerts different degrees of toxic effects on the swimming capability of Daphnia magna under otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 202. The testing aim was to determine a EC50 after 48 hours of exposure to D. magna. The stock solution used for the exposure assessment was prepared by dissolving colourless dense liquid in acetone. Daphnids were exposed to chemical in 50 ml beakers in a volume of 25 ml of liquid solution containing both the chemical and media as specified in OECD 202. The beakers were placed in a temperature controlled room at 20±1 degrees Celsius. The D. magna (age ≤24) used for the test. The animals were exposed to stock solution prepared by adding dense colourless liquid in acetone. There were 5 Daphnia per test vessels and 4 replicates per concentration. The positive control/reference substance used in the teste showed an expected result and gave a EC50 that corresponded to previous exposures with this chemical in D. magna. Immobilisation effects in D. manga by chemical exposure was evident after 48 hours in exposure concentrations above 1 mg/L (i.e. EC20). The EC50 was defined as a concentration that immobilizes 50% of the exposed D. magna. The % of immobilization in D. magna after 48 hours of exposure to test chemical were used in a nonlinear regression by Graphpad Software Prism 4.0 (San Diego, US). Based on the immobility of daphnia magna by the chemical exposure for 48 hrs, the EC50 was determine to be 2.59 mg/l (95% C.I: 1.81-3.7 mg/L). 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 classification criteria.

 

Long term toxicity to aquatic invertebrate:

The test substance was used to evaluate its adverse effects to Daphnia magna. The test was conducted in accordance with the OECD 211 guidleines, for the period of 21 days. The test solutions was prepared in the ADAMS medium, the test concentration chosen for the exposure was 0.25 - 4 mg/L, with the geometric series of 2. the test was performed in semi static conditions, wherein the test chemical was renewed every alternate day, of the exposure period. The EC10 was calculated to be 0.148±0.85 mg/L. Based on the Effective concentration it can be classified into chronic 2 category of CLP criteria.

 

 

Toxicity to algae and cyanobacteria:

Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance according to the OECD Guideline 201. The stock solution (32 g/L) was prepared by dissolving colourless dense liquid in acetone. 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, 2, 4, 8.5, 16, 32 and 68 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 (ErC50) for the test substance in Desmodesmus subspicatus was determined to be 47.0 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 3 as per the CLP classification criteria.

 

Toxicity to microorganisms:

Data available for the test chemical and read across chemicals selected on the basis of structural and functional similarity has been reviewed to determine its effect on microorganism. The studies are as mentioned below:

 

In the first study toxicity of test material was evaluated for microorganisms by disc diffusion method and the minimum inhibition concentration was evaluated using broth dilution method. The organisms used for the test were S. aureus, B. cereus , E. coli , P. aeruginosa. The turbidity of microorganisms was adjusted to 0.5. The inoculum were 1×1081×106CFU/ml for bacteria and fungi respectively. Inoculate was swabbed on Muller Hinton Agar. Sterile blank discs impregnated with 3, 5, 10 μl of oil in 10 μl of dimethyl sulfoxide (DMSO) were used and put on cultured plates. Disc containing DMSO and antibiotics were used as control. The minimum inhibition concentration of test material on bacteria S. aureus, B. cereus , E. coli , P. aeruginosa was observed to be > 8 mg/l.

 

Above study was supported by the second from secondary source. Toxicity of test material was evaluated for microorganisms by disc diffusion method and the minimum inhibition concentration was evaluated using broth dilution method. The organisms used for the test were S. aureus, B. cereus , E. coli , P. aeruginosa. The turbidity of microorganisms was adjusted to 0.5. The inoculum were 1×1081×106CFU/ml for bacteria and fungi respectively. Inoculate was swabbed on Muller Hinton Agar. Sterile blank discs impregnated with 3, 5, 10 μl of oil in 10 μl of dimethyl sulfoxide (DMSO) were used and put on cultured plates. Disc containing DMSO and antibiotics were used as control. The minimum inhibition concentration of test material on bacteria S. aureus, B. cereus, E. coli , P. aeruginosa was observed to be 1 mg/l.

 

The test chemical is likely to be toxic to microorganism atleast in the concentration range of 1 to 8 mg/l.

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