Sulisobenzone

Endpoint summary

Description of key information

Short term toxicity to fish:

Short term toxicity to fish Leuciscus idus for 96 hrs for the test chemical was examined in accordance with method closely followed the guideline of DIN 38 412 "Test procedure with water organisms (group l). General information The planning, implementation and evaluation of biological test procedures (l1) "and" determination of the effect of water on ingredients In fish - fish test (l15) ", june 1982 using a static procedure.The fish were kept in a flow-through tank in tap water cleaned by active carbon and aerated with oil-free air .Nominal concentrations used were 46.4, 100, 215, 464 mg/L. Short term toxicity to fish Leuciscus idus for 96 hrs fo test chemical examined at effect level as  NOEC: 215 mg/L and LC50: > 215 - <464 mg/L and LC100: 416 mg/L this all values indicate that the chemical dose not exhibit short term toxicity to fish and can be considered to be not classiffied as per CLP criteria.

Long term toxicity to fish:

The 14-days fish experiments were conducted using juvenile, sexually undifferentiated fathead minnows (Pimephales promelas), between 2

and 3 months of age and with a total body length between 19 and 27 mm . Fish were held in well-aerated reconstituted tap water medium.

The studies were conducted using a 24-h static-renewal procedure with daily renewal of total aquaria water. For exposure, 10 randomly selected fish were each placed in stainless-steel tanks (10 liter) and exposed to individual UV filters for 14 days. Long term toxicity (14 d) value of test material to Pimephales promelas examined of NOEC:1.048 mg/l (on the basis of body size) and NOEC: >4.897 mg/l (on the basis of mortality). Based on the above effect concentration it is considered to be non - hazardous to aquatic environment and can not be classified as per CLP criteria .

Short-term toxicity to aquatic invertebrates:

Daphnia sp., Acute Immobilization Test according to OECD Guideline 203 was conducted for test material. The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 200 mg of the test substance in 200 ml of ADaM’s media. Achieving test concentrations of 6.25 mg/L,12.5 mg/L,25 mg/L,50 mg/L, and 100 mg/L, respectively. Daphnia magna were exposed to these concentration for 48 hours. The median lethal concentration (EC0) for test material on Daphnia magna in a 48 hours study on the basis of immobilization effect was found to be 100 mg/l. Thus, on the basis of this EC0 value and according to CLP criteria for aquatic classification of the substance, it is concluded that the substance, does not exhibit short term toxicity to Daphnia.

Based on the EC50, it can be consider that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria.

Long term toxicity to aquatic invertebrate:

21 d reproduction study performed according to OECD guideline 211. Renewal of medium took place every 48 h, and the offsprings produced by each parent animal were counted daily. The exposure concentration was in the range of a tenth of the acute toxicity value. No adverse effects on either the number or sex of offsprings, or body size were observed after exposure to five concentrations each .Long term toxicity (21 d) value of sulisobenzone (benzophenone-4) to Daphnia magna resulted in a no observed effect concentration ( NOEC) ranging between 0.128–5 mg/L. Based on the above effect concentration it is considered to be non - hazardous to aquatic environment and can not be classified as per CLP criteria .

Toxicity to aquatic algae and cyanobacteria:

The study was designed to assess the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test).

Test was carried out in 100mL conical flasks which were carefully autoclaved and sterilized.The test item Sulisobenzone was prepared by adding 100 mg of test item in 500 ml of BBM to get the final concentration of 200 mg/L. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit.

The test concentration chosen for the study were 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L,200mg/L. The test concentrations were prepared using stock solution of the test item using mineral media.

For the assessment of algal growth, the test was conducted in replicates. The control flask was maintained in triplicates as recommended in the OECD guideline and the test concentration were selected in geometric series which were maintained in duplicates. To obtain a quantitative concentration-response relationship by regression analysis, a linearizing transformation of the response data into probit was performed. Using the same, effective concentration (EC) were determined.

Algal growth was calculated daily by counting the cells microscopically with the help of haemocytometer. For microscopic observations the cultures were observed daily with the help of a microscope to verify a normal and healthy appearance of the algal culture and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test item). Apart from this, the cell count of each test vessel was also noted with the help of a microscope and haemocytometer. By spectrophotometer the absorbance values of each test vessel and control vessel was noted at 680nm.The BBM was taken as blank for both control and test vessels. The absorbance value of each vessel was in line with the average specific growth rate.

As per OECD 201, the biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72 hr test period. This corresponds to a specific growth rate of 0.92 per day. Thus, the observed specific growth rate in the control cultures during the experiment was 0.358 per day. Secondly the mean coefficient of variation for section by section specific growth rates (days 0-1, 1-2 & 2-3, for 72 hr tests) in the control cultures must not exceed 35%. Thus, the observed mean coefficient of variation in the control cultures during the experiment was 33.42%. Thirdly the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 10%. Thus, the observed coefficient of variation of average specific growth rates during the experiment in control cultures was 8.26%. Hence, the test is considered valid as per OECD guideline, 201

After 72 hours of exposure to test item to various nominal test concentrations, EC50 was determine to be 109.55 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was not hazardous and can be consider to be not classified as per the CLP classification criteria.

Toxicity to microorganisms:

20 , 50 and 80% luminescence inhibition of test organism Vibrio fischeri NRRL-B-11177 was evaluted after 30 minutes of exposure of test material. The luminescence of bacteria was measured on a LUMIStox 300 luminometer . The 30 min inhibitory concentrations, IC20, IC50 and IC80 values were calculated using a log-linear regression analysis. The 30 min inhibitory concentrations, IC20, IC50 and IC80 was observed to be 67.3 , 301 , 1350 mg/l respectively.

Additional information

Short term toxicity to fish:

The effect of test material was observed on fish considering two experimental reports as mentioned below:

Short term toxicity to fish Leuciscus idus for 96 hrs for the test chemical was examined in accordance with method closely followed the guideline of DIN 38 412 "Test procedure with water organisms (group l). General information The planning, implementation and evaluation of biological test procedures (l1) "and" determination of the effect of water on ingredients In fish - fish test (l15) ", june 1982 using a static procedure.The fish were kept in a flow-through tank in tap water cleaned by active carbon and aerated with oil-free air .Nominal concentrations used were 46.4, 100, 215, 464 mg/L. Short term toxicity to fish Leuciscus idus for 96 hrs fo test chemical examined at effect level as  NOEC: 215 mg/L and LC50: > 215 - <464 mg/L and LC100: 416 mg/L this all values indicate that the chemical dose not exhibit short term toxicity to fish.

In another 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).The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 12.5 mg , 25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L & 400 mg/L of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 3.125 mg/L, 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 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. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be 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 3 as per the CLP classification criteria. The test substance is redialy degradable in aquatic environment and has low chances to be hazardous to aquatic environment . Hence , it can be considered to be not classified as per CLP criteria.

Long term toxicity to fish:

The 14-days fish experiments were conducted using juvenile, sexually undifferentiated fathead minnows (Pimephales promelas), between 2

and 3 months of age and with a total body length between 19 and 27 mm . Fish were held in well-aerated reconstituted tap water medium.

The studies were conducted using a 24-h static-renewal procedure with daily renewal of total aquaria water. For exposure, 10 randomly selected fish were each placed in stainless-steel tanks (10 liter) and exposed to individual UV filters for 14 days. Long term toxicity (14 d) value of test material to Pimephales promelas examined of NOEC:1.048 mg/l (on the basis of body size) and NOEC: >4.897 mg/l (on the basis of mortality). Based on the above effect concentration it is considered to be non - hazardous to aquatic environment and can not be classified as per CLP criteria .

Short-term toxicity to aquatic invertebrates:

Effect of test material was observed on aquatic invertebrate in an experimental report along with the data from peer reviewed journal.

In experimental report ,daphnia sp., Acute Immobilization Test according to OECD Guideline 203 was conducted for test material. The test substance was soluble in water. Therefore, the stock solution was prepared by dissolving 200 mg of the test substance in 200 ml of ADaM’s media. Achieving test concentrations of 6.25 mg/L,12.5 mg/L,25 mg/L,50 mg/L, and 100 mg/L, respectively. Daphnia magna were exposed to these concentration for 48 hours. The median lethal concentration (EC0) for test material on Daphnia magna in a 48 hours study on the basis of immobilization effect was found to be 100 mg/l. Thus, on the basis of this EC0 value and according to CLP criteria for aquatic classification of the substance, it is concluded that the substance, does not exhibit short term toxicity to Daphnia.

Based on the EC50, it can be consider that the chemical was not toxic and can be consider to be not classified as per the CLP classification criteria.

In peer reviewed journal ,the acute toxicity was determined according to the OECD guideline 202 (48 h acute immobilization assay). After 48 h, motile and immobilized D. magna were counted. The LC50 value in Daphnia magna was observed to be 50 mg/l after 48 h.Test substance is redialy degradable in aquatic environment , hence test substance is considered to be not classified as per CLP criteria.

Long term toxicity to aquatic invertebrate:

21 d reproduction study performed according to OECD guideline 211. Renewal of medium took place every 48 h, and the offsprings produced by each parent animal were counted daily. The exposure concentration was in the range of a tenth of the acute toxicity value. No adverse effects on either the number or sex of offsprings, or body size were observed after exposure to five concentrations each .Long term toxicity (21 d) value of sulisobenzone (benzophenone-4) to Daphnia magna resulted in a no observed effect concentration ( NOEC) ranging between 0.128–5 mg/L. Based on the above effect concentration it is considered to be non - hazardous to aquatic environment and can not be classified as per CLP criteria .

Toxicity to aquatic algae and cyanobacteria:

The study was designed to assess the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test).

Test was carried out in 100mL conical flasks which were carefully autoclaved and sterilized.The test item Sulisobenzone was prepared by adding 100 mg of test item in 500 ml of BBM to get the final concentration of 200 mg/L. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit.

The test concentration chosen for the study were 6.25mg/L,12.5mg/L,25mg/L,50mg/L,100mg/L,200mg/L. The test concentrations were prepared using stock solution of the test item using mineral media.

For the assessment of algal growth, the test was conducted in replicates. The control flask was maintained in triplicates as recommended in the OECD guideline and the test concentration were selected in geometric series which were maintained in duplicates. To obtain a quantitative concentration-response relationship by regression analysis, a linearizing transformation of the response data into probit was performed. Using the same, effective concentration (EC) were determined.

Algal growth was calculated daily by counting the cells microscopically with the help of haemocytometer. For microscopic observations the cultures were observed daily with the help of a microscope to verify a normal and healthy appearance of the algal culture and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test item). Apart from this, the cell count of each test vessel was also noted with the help of a microscope and haemocytometer. By spectrophotometer the absorbance values of each test vessel and control vessel was noted at 680nm.The BBM was taken as blank for both control and test vessels. The absorbance value of each vessel was in line with the average specific growth rate.

As per OECD 201, the biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72 hr test period. This corresponds to a specific growth rate of 0.92 per day. Thus, the observed specific growth rate in the control cultures during the experiment was 0.358 per day. Secondly the mean coefficient of variation for section by section specific growth rates (days 0-1, 1-2 & 2-3, for 72 hr tests) in the control cultures must not exceed 35%. Thus, the observed mean coefficient of variation in the control cultures during the experiment was 33.42%. Thirdly the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 10%. Thus, the observed coefficient of variation of average specific growth rates during the experiment in control cultures was 8.26%. Hence, the test is considered valid as per OECD guideline, 201

After 72 hours of exposure to test item to various nominal test concentrations, EC50 was determine to be 109.55 mg/l graphically and through probit analysis. Based on the EC50, it can be concluded that the chemical was not hazardous and can be consider to be not classified as per the CLP classification criteria.

Toxicity to microorganisms:

20 , 50 and 80% luminescence inhibition of test organism Vibrio fischeri NRRL-B-11177 was evaluted after 30 minutes of exposure of test material. The luminescence of bacteria was measured on a LUMIStox 300 luminometer . The 30 min inhibitory concentrations, IC20, IC50 and IC80 values were calculated using a log-linear regression analysis. The 30 min inhibitory concentrations, IC20, IC50 and IC80 was observed to be 67.3 , 301 , 1350 mg/l respectively.