Benzyl isovalerate

Endpoint summary

Description of key information

Short term toxicity to fish:

Acute fish toxicity study of 96 hrs was conducted to assess toxic effects of test chemical and the results were predicted. The study was based on the effects of the test compound on fish in a static fresh water system. The lethal concentration (LC50) for chemical was predicted to be 7.390 mg/l on the basis of mortality effects. Thus, based on lethal concentration, it can be concluded that test chemical considered as toxic to fish and thus can be classified as aquatic chronic category 2. Since the test chemical is readily biodegradable in nature, it can be considered as non-toxic to aquatic organisms and thus cannot be classified as hazardous as per the CLP criteria.

Short term toxicity to aquatic invertebrate:

Aim of this study was to access the short term toxicity of test chemical to aquatic invertebrate 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 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 reconstituted test water. Test conducted on different test concentration 0, 0, 6, 10, 17, 29 and 50 mg/l. 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 15.5 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and classified as per the CLP criteria. But as the chemical was readily biodegradable in water thus it concluded to be non-hazardous to aquatic invertebrates and cannot be classified as per the CLP criteria.

 

Toxicity to aquatic algae and cyanobacteria:

Aim of this study was to evaluate the nature of test chemical when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD Guideline 201 (Alga, Growth Inhibition Test). The stock solution 200 mg/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 inoculum culture. 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 (ErC50) for the test substance benzyl 3-methylbutanoate in algae was determined to be 59.9 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 3 category as per the CLP criteria. But as the chemical was readily biodegradable in water thus it concluded to be non-hazardous to aquatic invertebrates and cannot be classified as per the CLP criteria.

Toxicity to microorganism:

WoE 2: Based on the growth inhibition of microorganisms, LOEC was determine to be 236 mg/l for test material in Entosiphon sulcatum when it was exposed for 72 h.

WoE 3: The 50% Effective concentration of test chemical to Photobacterium phosphoreum was determined to be 3.02 mg/l based on viability and sensitivity of bacteria.

Additional information

Short term toxicity to fish:

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

Acute fish toxicity study of 96 hrs was conducted to assess toxic effects of test chemical and the results were predicted. The study was based on the effects of the test compound on fish in a static fresh water system. The lethal concentration (LC50) for chemical was predicted to be 7.390 mg/l on the basis of mortality effects. Thus, based on lethal concentration, it can be concluded that test chemical considered as toxic to fish and thus can be classified as aquatic chronic category 2. Since the test chemical is readily biodegradable in nature, it can be considered as non-toxic to aquatic organisms and thus cannot be classified as hazardous as per the CLP criteria.

 

Prediction done using average value of both models i.e Leadscope and SciMatics SciQSAR model and the result were predicted in Battery model. Based on the Danish (Q)SAR Database, the 96 hours LC50 was estimated to be 3.96 mg/l on Pimephales promelas for substance with mortality effects. Thus based on this value it can be concluded that the substance can be classified as toxic. But as the chemical was readily biodegradable in water thus chemical consider to be nontoxic to fish and not classified as per the CLP classification criteria.

 

Above data was supported by the third study from experimental report. 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). Therefore the stock solution prepared as 2g /4 liter, with the concentration of 500 mg/L, and was kept for 4 hours stirring. After the completion of stirring, the sample was run through HPLC system for getting the actual water solubility of the test substance. The actual solubility obtained was 122.66 mg/L. From this stock solution further test concentrations were prepared for achieving test concentrations of 6.25mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L, respectively. Potable water (passed through reverse osmosis system) was used and Zebra Fish Danio rerio were exposed to this 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 . Based on the LC50, it can be consider that the chemical was toxic and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria. But on the basis of readily biodegradability of test chemical, chemical consider to be nontoxic and not classified as per the CPL classification criteria.

 

In experimental study from peer reviewed journal short term toxicity study to Cyprinus carpio was carried out for 44 hrs. Test was performed under flow through conditions. After exposure of test animal to different dose conc. i.e, at 68, 130 and 146 mg/l of test chemical. Glass-fronted, fiberglass tanks were used in the study, no effects were observed on the test animal. Thus, the NOEC value was consider to be 146 mg/l respectively.

 

This fifth experimental study also supports the nontoxic nature of test chemical. Study was designed to access the toxic effects of the test compound on the Zebra fish (Danio rerio). Bowl aquaria containing 4 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. 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 were and test fish were exposed to 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L concentrations for 96 hours. The test substance was moderately soluble in water. Initially, stock solution was prepared by dissolving 1ml of the test substance in 1 liter of potable water (passed through reverse osmosis system) with 24 hrs of continuous stirring. From this stock solution, further test concentration was prepared for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively.  The median lethal concentration (LC50) for test chemical on Danio rerio in a 96 hours study on the basis of mortality effect was determine to be >12.5 mg/L. Thus, on the basis of this LC50 value and according to CLP criteria for aquatic classification of the substance, it is concluded that the substance, is classified under aquatic chronic 3 category and exhibits short term toxicity to fish. But as the chemical was readily biodegradable in water, thus on that basis, chemical consider to be nontoxic and not classified as per the CLP classification criteria.

 

Thus based on the all above studies, chemical consider to be nontoxic and not classified as per the CLP classification criteria.

 

Short term toxicity to aquatic invertebrate:

Aim of this study was to access the short term toxicity of test chemical to aquatic invertebrate 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 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 reconstituted test water. Test conducted on different test concentration 0, 0, 6, 10, 17, 29 and 50 mg/l. 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 15.5 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic invertebrates and classified as per the CLP criteria. But as the chemical was readily biodegradable in water thus it concluded to be non-hazardous to aquatic invertebrates and cannot be classified as per the CLP criteria.

 

Toxicity to aquatic algae and cyanobacteria:

Aim of this study was to evaluate the nature of test chemical when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD Guideline 201 (Alga, Growth Inhibition Test). The stock solution 200 mg/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 inoculum culture. 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 (ErC50) for the test substance benzyl 3-methylbutanoate in algae was determined to be 59.9 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Based on the EC50 value, indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 3 category as per the CLP criteria. But as the chemical was readily biodegradable in water thus it concluded to be non-hazardous to aquatic invertebrates and cannot be classified as per the CLP criteria.

Toxicity to microorganism:

Various studies available for the test chemical were reviewed to determine the toxic nature of test chemical on the growth and other activity of microorganisms. The studies are as mentioned below:

 

In the first experimental study from peer reviewed journal toxicity to microorganism study for test material the biological effect was evaluated by using analogous methods of the cell multiplication inhibition test. For maintenance of the test strain of Entosiphon sulcatum continuously inoculate the expected required number of stock cultures at 72 h or 96 h intervals. For this, mix 8 ml of stock solution I and 8 ml of sterile double-distilled water in 300-ml Erlenmeyer flasks stoppered with metal caps and add 2 ml of a stock culture and 2 ml of the adjusted bacterial suspension for Entosiphon stock cultures. Store stock cultures at 25°C. Before inoculation control the flasks containing the stock cultures by means of an inverse microscope. Pretreatment of the bacteria used for feeding preliminary cultures of Entosiphon sulcatum does not differ from that of bacteria used for feeding the stock cultures. Keep preliminary cultures at 25°C over 72 h before using them for inoculation of the test cultures. Before inoculation control each flask containing the preliminary cultures by means of an inverse microscope. The toxicity threshold (TT) of test material was determined to be 236 mg/l for Entosiphon sulcatum. Therefore LOEC was considered to be 236 mg/l for 72h in Entosiphon sulcatum.

 

In the second study toxicity were measured. Photobacterium phosphoreum was test organism used to determine the toxicity of test material done by Microtox test method. The time period was 5 to 30 minute at 15 degree C and pH range from 5 to 9. After exposure, the 50% Effective concentration value for 1 test chemical to Photobacterium phosphoreum was determined to be 3.02 mg/l based on viability and sensitivity of bacteria. As the chemical were readily biodegradable thus degraded faster and consider to be nontoxic.

Thus based on the above all studies and effects observation, chemical consider to be nontoxzic and not classified as per the CLP classification criteria.