2'-acetonaphthone

Endpoint summary

Description of key information

Short-Term Toxicity to Fish

This study was conducted as per OECD 203 (2019) to assess the acute toxicity effects of test chemical on zebrafish (Danio rerio) following exposure up to 96 h under static condition. Juvenile fish of same age and normal in appearance were used in this (originate from same source and population). The average length and weight (10 fish) were observed 1.4 cm and 0.012g, respectively. Fish were fed (commercial fish food) daily during acclimatization. Photoperiod and light intensity were maintained such as16 h light- 8 h dark, 732-804 Lux during experiment. Hardness of water was measured once during acclimatization and found to be 165 mg CaCO3/L, temperature, pH and dissolve oxygen were maintained between 22.1-22.7 °C,6.4-7.1,6.8-8.5 mg/L, respectively, throughout the test. Fish were acclimatized for 7 days prior dosing. Natural water was used as dilution control and acetone in natural water (500µL/5000 mL conc) was used as vehicle control and the same concentration was used for formulation of test chemical. The study was initiated with a range finding test by using following concentrations of 0 (control), 0 (vehicle control), 2.35, 4.23, 7.61, 13.71, 24.67 mg/L. During range finding test, no mortality or abnormality was found in control groups and 2.35, 4.23, 7.61 mg/L conc., but 100% mortality were found in 13.71, 24.67 mg/L concentrations. Hence, a main study was conducted using 0 (control), 0 (vehicle control), 6.61, 7.93, 9.52, 11.42, 13.71 mg/L concentrations. No mortality or abnormality was found in control groups and 6.61, 7.93 mg/L concentration but abnormal behavioural responses like head shaking, loss of buoyancy of control, corkscrew swimming were observed in 9.52, 11.42, 13.71 mg/L conc. 7 fish were used/concentration during range finding and main study. HPLC method was used for method validation and active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 h and 96 h avg. recovery for 6.61, 9.52, 13.71 mg/L conc were 96.80%, 95.72%, 95.72% & 96.92%, 95.83%, 95.58%, respectively, were found in acceptable range. The test is valid as all the validity criteria are fulfilled: No mortality in control or vehicle control were found throughout the 96-h test duration; Dissolve oxygen conc. was maintained above 60% in all test vessels throughout the test; The recovery active ingredient content was found between 80-120% up to 96h. The 96 h LC50 of test chemical to zebrafish (Danio rerio) was determined to be 10.44 mg/L with 95% confidence upper and lower limit were found to be 10.79 and 10.09 mg/L, respectively. 96h LC50 value with 95% confidence limits (upper limit, lower limit) were calculated by probit analysis using NCSS Software 2019, version 19.0.3. Hence, as per CLP classification category, the test chemical can be categorized as aquatic chronic category 3.

Long-Term Toxicity to Fish

The 28 days chronic toxicity to fish was predicted by using the ECOSAR 1.11 version. The predicted NOEC value for test chemical was 4.219 mg/L. This NOEC suggests that test chemical could not exhibit long term toxicity to fish and thus can be considered to not classified according to CLP criteria.

Short-Term Toxicty to Aquatic Invertebrates

This study was designed (as per OECD 202, adopted in 2004) to assess the acute toxicity of test chemical following exposure of daphnids up to 48h by static method.The brood daphnids were acclimatized 48 hours prior to the test item exposure. Less than 24 h old daphnids were collected from the acclimatized gravid females and exposed to the test item. After exposure on day 0, daphnids were observed for immobilization at 24 and 48 h. Natural water was used as control, acetone with natural water was used as vehicle control and the same was used for test item formulation and test medium (15 µL acetone/150 mL natural water concentration was used). 25 mL glass beakers having a solution volume of 20 mL were used in the test. A range finding test (2 replicates/concentration having 5 daphnids/replicate) with the test concentrations of 0 (control), 0 (vehicle control), 1.5, 3, 6, 12, 24 mg/L was done prior to main study. No immobilization or abnormality was found in control groups and 1.5, 3, 6.0 mg/L conc., but 20% and 100% mortality were found in 12, 24 mg/L conc, respectively. Hence, a main study (using a spacing factor of 1.4) was conducted using 0 (control), 0 (vehicle control), 6.0, 8.4, 11.8, 16.5, 23.0 mg/L concentrations. 4 replicates/concentration having 5 daphnids/replicate was used for the main study. Normal behavioural response and no immobilization (0% mortality) were observed up to 48 h followed by control groups and 6.0, 8.4 mg/L but 20%, 35% and 85% mortality were observed in the test concentrations of 11.8, 16.5, 23.0 mg/L, respectively. HPLC method was used for method validation and active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 h and 48 h avg. recovery for 6.0, 11.8, 23.0 mg/L conc were 95.80%, 95.30%, 95.70% & 93.93%, 95.13%, 95.52%, respectively, were found in acceptable range. Hence the results were based on nominal concentration, since the deviation in the initial measured concentration didn’t exceed 20%. Environmental parameters such as pH (7.3-7.9), temperature (20-21 °C), dissolve oxygen (6.8-7.2 mg/L), hardness (165 mg CaCO3/L), conductivity (0.227 µS/cm), photoperiod (16 h light- 8 h dark) and light intensity (1337-1346 Lux) was maintained in acceptable range throughout the test. Feed was not provided during the test. The 24-h and 48-h EC50 of test chemical to daphnid,Daphnia magnaare 20.34 and 17.13 mg/L, respectively. The 48-h EC50 of reference item (Potassium dichromate) to daphnid,Daphnia magna(found to be in acceptable range) is 0.690 mg/L. Hence, the results of the test with reference item establish the acceptability of the test system response, test procedures followed and results obtained with test item. 48h EC50 value with 95% confidence limits (upper limit, lower limit) were calculated by probit analysis using NCSS Software 2019, version 19.0.3. Hence, as per CLP classification category, the test chemical can be categorized as aquatic chronic category 3.

Long-Term Toxicty to Aquatic Invertebrates

The 21 days chronic toxicity to fish (ChV) was predicted using the ECOSAR 1.11 version. The predicted ChV value for test chemical was 2.76 mg/L. Based on the ChV chemical cannot be classified as per CLP category

Toxicity to Aquatic Algae and Cyanobacteria

This study was conducted as per OECD 201 (2011) to assess the acute toxicity effects of test chemical on the growth of green alga Pseudokirchneriella subcapitata(ATCC)following exposure of alga up to 72 h under static condition. Test system was initially procured from “American Type Culture Collection”, Chromachemie Laboratory Private Limited, later test system was sub-cultured (using OECD growth medium, OECD 201) in the test facility. 250 mL sterilized conical flasks covered with cap were used in the study. The test item was found to be soluble in acetone. Hence, acetone (concentration <0.1mL/L) with OECD growth medium was used as test medium. An inoculum culture in the test medium was prepared 3-4 days prior to test and culture conditions were maintained same as the test conditions. A range finding study (3 replicates/ concentration, 6 replicates for control group)with the test concentrations of1.5, 3, 6, 12, 24 mg/L were tested along with a control (0), vehicle control (0) group, prior to the main study. The inhibition in growth rate (0%, 0%, 3.04%, 22.18%, 31.09%, 87.92%, 100%) and yield (0%, 0%, 11.69%, 60.46%, 73.02%, 98.85%, 100%) were observed at 72 h in the test concentrations of 0, 0, 1.5, 3, 6, 12, 24 mg/L. Based on the results of range finding test following concentrations of 0 (control), 0 (vehicle control), 0.8, 1.6, 3.2, 6.4, 12.8 mg/L were selected for the main study(3 replicates/ concentration, 6 replicates for control groups)as significant changes (inhibition algal growth rate) were observed in treatment groups during 72h test period. The inhibition in growth rate (0%, 0%, 1.78%, 6.68%, 24.76%, 33.68%, 90.82%) and yield (0%, 0%, 7.17%, 24.49%, 65.09%, 76.32%, 99.19%) were observed during 72h test period in the test concentrations of 0 (control), 0 (vehicle control), 0.8, 1.6, 3.2, 6.4, 12.8 mg/L. A reference standard (Potassium dichromate) was tested to ensure the authenticity of the test in the lab. And the inhibition of growth rate (ErC50-72h) and yield (EyC50-72h) were found to be 0.620 mg/L & 0.334 mg/L, respectively; and were found to be in acceptable range. The initial cell density was 10000 cells/mL.The algal cells in the control and vehicle control increased by 64.25, 62.58 times (>16 times) of the initial cell count during the 72-h exposure period, respectively. The mean coefficient of variation for section-by-section growth rate for the control cultures over the test period (0-72-h) was 32.8% (<35%). The coefficient of variation of average specific growth rate was 0.6% (<7%). Hence, fulfilling the all the validity criteria of the test. HPLC method was used for method validation and active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 hr and 72 hr avg. recovery for 0.8, 3.2, 12.8 mg/L conc were 94.34%, 94.44%, 95.84% & 94.11%, 94.59%, 96.11%, respectively, were found in acceptable range. Continuous light having and average light intensity of 6766-6794 Lux, pH of 7.6-8.1, temperature 21.7-23.9 °C along with a shaking speed of 110 RPM were provided and maintained for the test system throughout the test. 72h EC50 (growth rate and yield) values with 95% confidence limits were calculated by probit analysis. NOEC and LOEC of growth rate and yield were calculated by one way ANOVA (Kruskal-Wallis-Comparison Z-Value Test) using NCSS Software 2019, version 19.0.3. The cells were counted using Haemocytometer under illumination of the microscope at 24, 48, and 72 h after inoculation. 72-h EC50 of test item to alga on growth rate and yield are 6.19, 2.73 mg/L, respectively. 72-h LOEC of test item to alga on growth rate and yield is 3.2 mg/L. 72-h NOEC of test item to alga on growth rate and yield is 1.6 mg/L. Thus based on the outcomes test chemical was classified into aquatic chronic category 2 as CLP classification criteria.

Toxicity to Microorganisms

In different studies, the given test chemical, has been investigated for toxicity to microorganisms to a greater or lesser extent. The studies are summarized as below –

In this study both the test chemicals were studied to understand its effects on microorganisms, both the tests were performed in the static regime and the inhibitory growth concentration (IGC50) value was determined. In the first WoE experiment tenure was for 60 hours and the second WoE study was for 48 hours.

It was determined that the IGC for study 2 and study 3 is 60.61 and 17 mg/L respectively

Thus from the above effects chemical toxicity value ranges from 17 -60.61 mg/L

Additional information

Short-Term Toxicity to Fish

This study was conducted as per OECD 203 (2019) to assess the acute toxicity effects of test chemical on zebrafish (Danio rerio) following exposure up to 96 h under static condition. Juvenile fish of same age and normal in appearance were used in this (originate from same source and population). The average length and weight (10 fish) were observed 1.4 cm and 0.012g, respectively. Fish were fed (commercial fish food) daily during acclimatization. Photoperiod and light intensity were maintained such as16 h light- 8 h dark, 732-804 Lux during experiment. Hardness of water was measured once during acclimatization and found to be 165 mg CaCO3/L, temperature, pH and dissolve oxygen were maintained between 22.1-22.7 °C,6.4-7.1,6.8-8.5 mg/L, respectively, throughout the test. Fish were acclimatized for 7 days prior dosing. Natural water was used as dilution control and acetone in natural water (500µL/5000 mL conc) was used as vehicle control and the same concentration was used for formulation of test chemical. The study was initiated with a range finding test by using following concentrations of 0 (control), 0 (vehicle control), 2.35, 4.23, 7.61, 13.71, 24.67 mg/L. During range finding test, no mortality or abnormality was found in control groups and 2.35, 4.23, 7.61 mg/L conc., but 100% mortality were found in 13.71, 24.67 mg/L concentrations. Hence, a main study was conducted using 0 (control), 0 (vehicle control), 6.61, 7.93, 9.52, 11.42, 13.71 mg/L concentrations. No mortality or abnormality was found in control groups and 6.61, 7.93 mg/L concentration but abnormal behavioural responses like head shaking, loss of buoyancy of control, corkscrew swimming were observed in 9.52, 11.42, 13.71 mg/L conc. 7 fish were used/concentration during range finding and main study. HPLC method was used for method validation and active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 h and 96 h avg. recovery for 6.61, 9.52, 13.71 mg/L conc were 96.80%, 95.72%, 95.72% & 96.92%, 95.83%, 95.58%, respectively, were found in acceptable range. The test is valid as all the validity criteria are fulfilled: No mortality in control or vehicle control were found throughout the 96-h test duration; Dissolve oxygen conc. was maintained above 60% in all test vessels throughout the test; The recovery active ingredient content was found between 80-120% up to 96h. The 96 h LC50 of test chemical to zebrafish (Danio rerio) was determined to be 10.44 mg/L with 95% confidence upper and lower limit were found to be 10.79 and 10.09 mg/L, respectively. 96h LC50 value with 95% confidence limits (upper limit, lower limit) were calculated by probit analysis using NCSS Software 2019, version 19.0.3. Hence, as per CLP classification category, the test chemical can be categorized as aquatic chronic category 3.

Long-Term Toxicity to Fish

The 28 days chronic toxicity to fish was predicted by using the ECOSAR 1.11 version. The predicted NOEC value for test chemical was 4.219 mg/L. This NOEC suggests that test chemical could not exhibit long term toxicity to fish and thus can be considered to not classified according to CLP criteria.

Short-Term Toxicty to Aquatic Invertebrates

This study was designed (as per OECD 202, adopted in 2004) to assess the acute toxicity of test chemical following exposure of daphnids up to 48h by static method.The brood daphnids were acclimatized 48 hours prior to the test item exposure. Less than 24 h old daphnids were collected from the acclimatized gravid females and exposed to the test item. After exposure on day 0, daphnids were observed for immobilization at 24 and 48 h. Natural water was used as control, acetone with natural water was used as vehicle control and the same was used for test item formulation and test medium (15 µL acetone/150 mL natural water concentration was used). 25 mL glass beakers having a solution volume of 20 mL were used in the test. A range finding test (2 replicates/concentration having 5 daphnids/replicate) with the test concentrations of 0 (control), 0 (vehicle control), 1.5, 3, 6, 12, 24 mg/L was done prior to main study. No immobilization or abnormality was found in control groups and 1.5, 3, 6.0 mg/L conc., but 20% and 100% mortality were found in 12, 24 mg/L conc, respectively. Hence, a main study (using a spacing factor of 1.4) was conducted using 0 (control), 0 (vehicle control), 6.0, 8.4, 11.8, 16.5, 23.0 mg/L concentrations. 4 replicates/concentration having 5 daphnids/replicate was used for the main study. Normal behavioural response and no immobilization (0% mortality) were observed up to 48 h followed by control groups and 6.0, 8.4 mg/L but 20%, 35% and 85% mortality were observed in the test concentrations of 11.8, 16.5, 23.0 mg/L, respectively. HPLC method was used for method validation and active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 h and 48 h avg. recovery for 6.0, 11.8, 23.0 mg/L conc were 95.80%, 95.30%, 95.70% & 93.93%, 95.13%, 95.52%, respectively, were found in acceptable range. Hence the results were based on nominal concentration, since the deviation in the initial measured concentration didn’t exceed 20%. Environmental parameters such as pH (7.3-7.9), temperature (20-21 °C), dissolve oxygen (6.8-7.2 mg/L), hardness (165 mg CaCO3/L), conductivity (0.227 µS/cm), photoperiod (16 h light- 8 h dark) and light intensity (1337-1346 Lux) was maintained in acceptable range throughout the test. Feed was not provided during the test. The 24-h and 48-h EC50 of test chemical to daphnid,Daphnia magnaare 20.34 and 17.13 mg/L, respectively. The 48-h EC50 of reference item (Potassium dichromate) to daphnid,Daphnia magna(found to be in acceptable range) is 0.690 mg/L. Hence, the results of the test with reference item establish the acceptability of the test system response, test procedures followed and results obtained with test item. 48h EC50 value with 95% confidence limits (upper limit, lower limit) were calculated by probit analysis using NCSS Software 2019, version 19.0.3. Hence, as per CLP classification category, the test chemical can be categorized as aquatic chronic category 3.

Long-Term Toxicty to Aquatic Invertebrates

The 21 days chronic toxicity to fish (ChV) was predicted using the ECOSAR 1.11 version. The predicted ChV value for test chemical was 2.76 mg/L. Based on the ChV chemical cannot be classified as per CLP category

Toxicity to Aquatic Algae and Cyanobacteria

This study was conducted as per OECD 201 (2011) to assess the acute toxicity effects of test chemical on the growth of green alga Pseudokirchneriella subcapitata(ATCC)following exposure of alga up to 72 h under static condition. Test system was initially procured from “American Type Culture Collection”, Chromachemie Laboratory Private Limited, later test system was sub-cultured (using OECD growth medium, OECD 201) in the test facility. 250 mL sterilized conical flasks covered with cap were used in the study. The test item was found to be soluble in acetone. Hence, acetone (concentration <0.1mL/L) with OECD growth medium was used as test medium. An inoculum culture in the test medium was prepared 3-4 days prior to test and culture conditions were maintained same as the test conditions. A range finding study (3 replicates/ concentration, 6 replicates for control group)with the test concentrations of1.5, 3, 6, 12, 24 mg/L were tested along with a control (0), vehicle control (0) group, prior to the main study. The inhibition in growth rate (0%, 0%, 3.04%, 22.18%, 31.09%, 87.92%, 100%) and yield (0%, 0%, 11.69%, 60.46%, 73.02%, 98.85%, 100%) were observed at 72 h in the test concentrations of 0, 0, 1.5, 3, 6, 12, 24 mg/L. Based on the results of range finding test following concentrations of 0 (control), 0 (vehicle control), 0.8, 1.6, 3.2, 6.4, 12.8 mg/L were selected for the main study(3 replicates/ concentration, 6 replicates for control groups)as significant changes (inhibition algal growth rate) were observed in treatment groups during 72h test period. The inhibition in growth rate (0%, 0%, 1.78%, 6.68%, 24.76%, 33.68%, 90.82%) and yield (0%, 0%, 7.17%, 24.49%, 65.09%, 76.32%, 99.19%) were observed during 72h test period in the test concentrations of 0 (control), 0 (vehicle control), 0.8, 1.6, 3.2, 6.4, 12.8 mg/L. A reference standard (Potassium dichromate) was tested to ensure the authenticity of the test in the lab. And the inhibition of growth rate (ErC50-72h) and yield (EyC50-72h) were found to be 0.620 mg/L & 0.334 mg/L, respectively; and were found to be in acceptable range. The initial cell density was 10000 cells/mL.The algal cells in the control and vehicle control increased by 64.25, 62.58 times (>16 times) of the initial cell count during the 72-h exposure period, respectively. The mean coefficient of variation for section-by-section growth rate for the control cultures over the test period (0-72-h) was 32.8% (<35%). The coefficient of variation of average specific growth rate was 0.6% (<7%). Hence, fulfilling the all the validity criteria of the test. HPLC method was used for method validation and active ingredient analysis along with stability of the test item in the test medium. Test item was found to be stable in the test medium. The active ingredient content results were considered acceptable (80-120%) as during main study 0 hr and 72 hr avg. recovery for 0.8, 3.2, 12.8 mg/L conc were 94.34%, 94.44%, 95.84% & 94.11%, 94.59%, 96.11%, respectively, were found in acceptable range. Continuous light having and average light intensity of 6766-6794 Lux, pH of 7.6-8.1, temperature 21.7-23.9 °C along with a shaking speed of 110 RPM were provided and maintained for the test system throughout the test. 72h EC50 (growth rate and yield) values with 95% confidence limits were calculated by probit analysis. NOEC and LOEC of growth rate and yield were calculated by one way ANOVA (Kruskal-Wallis-Comparison Z-Value Test) using NCSS Software 2019, version 19.0.3. The cells were counted using Haemocytometer under illumination of the microscope at 24, 48, and 72 h after inoculation. 72-h EC50 of test item to alga on growth rate and yield are 6.19, 2.73 mg/L, respectively. 72-h LOEC of test item to alga on growth rate and yield is 3.2 mg/L. 72-h NOEC of test item to alga on growth rate and yield is 1.6 mg/L. Thus based on the outcomes test chemical was classified into aquatic chronic category 2 as CLP classification criteria.

Toxicity to Microorganisms

In different studies, the given test chemical, has been investigated for toxicity to microorganisms to a greater or lesser extent. The studies are summarized as below –

In this study both the test chemicals were studied to understand its effects on microorganisms, both the tests were performed in the static regime and the inhibitory growth concentration (IGC50) value was determined. In the first WoE experiment tenure was for 60 hours and the second WoE study was for 48 hours.

It was determined that the IGC for study 2 and study 3 is 60.61 and 17 mg/L respectively

Thus from the above effects chemical toxicity value ranges from 17 -60.61 mg/L

Based on the outcomes from the aquatic toxicity the test chemical can be classified into aquatic chronic 2 category.