Ethyltriphenylfosfonium bromide

Endpoint summary

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 soluble in water. Initially, stock solution was prepared by dissolving 1g of the test substance in 1 liter of potable water (passed through reverse osmosis system) with continuous stirring. From this stock solution, further test concentration was prepared for achieving test concentrations of 18 mg/L, 23.4 mg/L, 30.42 mg/L, 39.54 mg/L & 51.4 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 concentrations18 mg/L, 23.4 mg/L, 30.42 mg/L, 39.54 mg/L & 51.4 mg/l

LC50 was determine to be >39.42 mg/l . Based on the LC50,LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 51.4 mg/Lit can be consider that the chemical was toxic and can be consider to be classified aq aquatic chronic 3 as per the CLP classification criteria.

Short term toxicity to aquatic invertebrates:

Data available for the test chemical chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate .The studies are as mentioned below:

1) Short term toxicity to aquatic invertebrate Pomacea canaliculata (golden apple snail) was evaluated for the test material. Aquaria with dimensions of 50 cm x 25 cm x 35 cm were used for maintaining and breeding snails in the laboratory. The top of the aquarium was covered with nylon screen. Lamps were positioned 15 cm above the aquaria with a cycle of 16 hrs “on”, and 8 hrs “off’. The aquarium floor was covered with a layer of small pebbles, and was filled with water to a depth of 25 cm. Tap water reduced the thickness of the snail shell and the number of eggs, dechlorinated tap water caused the snails to secrete mucus. As a result, fresh aerated well water (pH 7.5, 26 °C) was provided in a continuous flow. Wild adult snails, with a diameter greater than 5 cm, were collected from the field. After copulation, fresh eggs were incubated and the young snails were hatched in 10-17 days. The young snails were fed sweet potato leaves in the aquarium for 65-70 days until maturity.The breeding was repeated and the third generation snails of 35-40 days old were used for toxicity testing.

 The bioassay was conducted in 500 mL glass bottles, each was tilled with thirty snails and 450 mL of water. The concentration of test compounds ranged from 0.5 to 250 mg/L (6 different concentrations, three replications). After 72 hrs exposure, the number  of dead snails were recorded. The snail was considered to be dead if there was no response to touch, the operculum shut tight, but with no resistance when the operculum was lifted, and the foot was out and swollen.The lethal effect concentration ( LC50) was determined to be 16.9 mg/l(linear regression (r²)= 2.441x + 8.742(0.942)). Based on the above effect concentration it can be considered that the test substance is toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

 

2)Short term toxicity to aquatic invertebrate Pomacea canaliculata (golden apple snail) was evaluated for the test material. Aquaria with dimensions of 50 cm x 25 cm x 35 cm were used for maintaining and breeding snails in the laboratory. The top of the aquarium was covered with nylon screen. Lamps were positioned 15 cm above the aquaria with a cycle of 16 hrs “on”, and 8 hrs “off’. The aquarium floor was covered with a layer of small pebbles, and was filled with water to a depth of 25 cm. Tap water reduced the thickness of the snail shell and the number of eggs, dechlorinated tap water caused the snails to secrete mucus. As a result, fresh aerated well water (pH 7.5, 26 °C) was provided in a continuous flow. Wild adult snails, with a diameter greater than 5 cm, were collected from the field. After copulation, fresh eggs were incubated and the young snails were hatched in 10-17 days. The young snails were fed sweet potato leaves in the aquarium for 65-70 days until maturity. The breeding was repeated and the third generation snails of 35-40 days old were used for toxicity testing.

 The bioassay was conducted in 500 mL glass bottles, each was tilled with thirty snails and 450 mL of water. The concentration of test compounds ranged from 0.5 to 250 mg/L (6 different concentrations, three replications). After 72 hrs exposure, the number  of dead snails were recorded. The snail was considered to be dead if there was no response to touch, the operculum shut tight, but with no resistance when the operculum was lifted, and the foot was out and swollen. The lethal effect concentration ( LC50) was determined to be 67.4 mg/l linear regression (r²)= 0.617 x + 8.382(0.932). Based on the above effect concentration it can be considered that the test substance is toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

3) Short term toxicity test for aquatic invertebrtae was performed for the test material for 24 h . The EC50 value was observed to be 52.6 mg/l , The EC0 was noted at 31.2 mg/l and the EC100 was observed in the concentration 125 mg/l.Based on the above effect concentartions it can be considered that test material was toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

Based on the above effect concentartions it can be considered that the test substance is toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

Long term toxicity to aquatic invertebrate:

Based on the prediction done using ECOSAR version 1.1, the long term toxicity on aquatic invertebrate was predicted for test substance . On the basis of effects observed in a static freshwater system, the NOEC value for the test substance is estimated to be0.163mg/l for aquatic invertebrate for 21 days of exposure, Based on the NOEC value it can be concluded that the test chemical can be considered as toxic to aquatic invertebrate at environmentally relevant concentrations and can be considered aquatic chronic 2 as per the CLP classification 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 solution was prepared in aseptic condition. The test substance was prepared by adding 60.75 mg of test substance in 250 ml of BBM to get the final concentration of 243 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 10⁴cells/ml. Care was taken to have a homogeneous solution for the experiment.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 substance  to various nominal test concentration, EC50 calculated from equation and graphically through probit analysis was found to be 30.9134 mg/L and 31.62 mg/L respectively.Based on the EC50, it can be concluded that the chemical was toxic to aqutic algae and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.

Toxicity to microorganism:

Data available for the test chemicals has been reviewed to determine the toxicity of microorganism .The studies are as mentioned below:

1) The effect of test material on Pseudomonas putida was observed for its Oxygen consumption. Pseudomonas putida, strain DSM 50026 were cultured on 25 g/l nutrient broth , 18 g/l Agar poured into slant-agar tubes. The stock solution was prepared with Tween 80 as dispersant and was stirred for 17 hours, resulting in a white suspension. The further dilutions were prepared from the stock solution while stirring. The nominal test concentration used were 156.25, 312.5, 625, 1250, 2500, 10000 mg/l. The effect concentartion EC50 was after 30 min of eposure to test organism Pseudomonas putida was observed to be >10000 mg/l. Based on the above effect concentration it can be considered that test substance is not toxic for the microorganisms.

2) Effect of test material on bio luminescence property of Photobacterium sp. was evaluated after 15 min of exposure of test material. The EC50 value of test material on Photobacterium sp. after 15 mins was observed to be 202 mg/l. Based on the effect concentration it can be concluded that test material has no adverse effect on microorganism.

Additional 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 soluble in water. Initially, stock solution was prepared by dissolving 1g of the test substance in 1 liter of potable water (passed through reverse osmosis system) with continuous stirring. From this stock solution, further test concentration was prepared for achieving test concentrations of 18 mg/L, 23.4 mg/L, 30.42 mg/L, 39.54 mg/L & 51.4 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 concentrations18 mg/L, 23.4 mg/L, 30.42 mg/L, 39.54 mg/L & 51.4 mg/l

LC50 was determine to be >39.42 mg/l . Based on the LC50,LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 51.4 mg/Lit can be consider that the chemical was toxic and can be consider to be classified aq aquatic chronic 3 as per the CLP classification criteria.

Short term toxicity to aquatic invertebrates:

Data available for the test chemical chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate .The studies are as mentioned below:

1) Short term toxicity to aquatic invertebrate Pomacea canaliculata (golden apple snail) was evaluated for the test material. Aquaria with dimensions of 50 cm x 25 cm x 35 cm were used for maintaining and breeding snails in the laboratory. The top of the aquarium was covered with nylon screen. Lamps were positioned 15 cm above the aquaria with a cycle of 16 hrs “on”, and 8 hrs “off’. The aquarium floor was covered with a layer of small pebbles, and was filled with water to a depth of 25 cm. Tap water reduced the thickness of the snail shell and the number of eggs, dechlorinated tap water caused the snails to secrete mucus. As a result, fresh aerated well water (pH 7.5, 26 °C) was provided in a continuous flow. Wild adult snails, with a diameter greater than 5 cm, were collected from the field. After copulation, fresh eggs were incubated and the young snails were hatched in 10-17 days. The young snails were fed sweet potato leaves in the aquarium for 65-70 days until maturity.The breeding was repeated and the third generation snails of 35-40 days old were used for toxicity testing.

 The bioassay was conducted in 500 mL glass bottles, each was tilled with thirty snails and 450 mL of water. The concentration of test compounds ranged from 0.5 to 250 mg/L (6 different concentrations, three replications). After 72 hrs exposure, the number  of dead snails were recorded. The snail was considered to be dead if there was no response to touch, the operculum shut tight, but with no resistance when the operculum was lifted, and the foot was out and swollen.The lethal effect concentration ( LC50) was determined to be 16.9 mg/l(linear regression (r²)= 2.441x + 8.742(0.942)). Based on the above effect concentration it can be considered that the test substance is toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

 

2)Short term toxicity to aquatic invertebrate Pomacea canaliculata (golden apple snail) was evaluated for the test material. Aquaria with dimensions of 50 cm x 25 cm x 35 cm were used for maintaining and breeding snails in the laboratory. The top of the aquarium was covered with nylon screen. Lamps were positioned 15 cm above the aquaria with a cycle of 16 hrs “on”, and 8 hrs “off’. The aquarium floor was covered with a layer of small pebbles, and was filled with water to a depth of 25 cm. Tap water reduced the thickness of the snail shell and the number of eggs, dechlorinated tap water caused the snails to secrete mucus. As a result, fresh aerated well water (pH 7.5, 26 °C) was provided in a continuous flow. Wild adult snails, with a diameter greater than 5 cm, were collected from the field. After copulation, fresh eggs were incubated and the young snails were hatched in 10-17 days. The young snails were fed sweet potato leaves in the aquarium for 65-70 days until maturity. The breeding was repeated and the third generation snails of 35-40 days old were used for toxicity testing.

 The bioassay was conducted in 500 mL glass bottles, each was tilled with thirty snails and 450 mL of water. The concentration of test compounds ranged from 0.5 to 250 mg/L (6 different concentrations, three replications). After 72 hrs exposure, the number  of dead snails were recorded. The snail was considered to be dead if there was no response to touch, the operculum shut tight, but with no resistance when the operculum was lifted, and the foot was out and swollen. The lethal effect concentration ( LC50) was determined to be 67.4 mg/l linear regression (r²)= 0.617 x + 8.382(0.932). Based on the above effect concentration it can be considered that the test substance is toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

3) Short term toxicity test for aquatic invertebrtae was performed for the test material for 24 h . The EC50 value was observed to be 52.6 mg/l , The EC0 was noted at 31.2 mg/l and the EC100 was observed in the concentration 125 mg/l.Based on the above effect concentartions it can be considered that test material was toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

Based on the above effect concentartions it can be considered that the test substance is toxic to aquatic invertebrate and can be classified as aquatic chronic 3 as per CLP criteria.

Long term toxicity to aquatic invertebrate:

Based on the prediction done using ECOSAR version 1.1, the long term toxicity on aquatic invertebrate was predicted for test substance . On the basis of effects observed in a static freshwater system, the NOEC value for the test substance is estimated to be0.163mg/l for aquatic invertebrate for 21 days of exposure, Based on the NOEC value it can be concluded that the test chemical can be considered as toxic to aquatic invertebrate at environmentally relevant concentrations and can be considered aquatic chronic 2 as per the CLP classification criteria.

Long term toxicity to fish:

Long term toxicity for test substance was evaluated using prediction for the test material and another data from authorative database:

1)Based on the prediction done using ECOSAR version 1.1, the long term toxicity on fish was predicted for test substance . On the basis of effects observed in a static fresh water system, the NOEC value for the substance is estimated to be0.156mg/l for fish for 28 days of exposure duration.Based on the NOEC value it can be concluded that the test chemical is toxic to fish at environmentally relevant concentrations and can be considered aquatic cronic 2 as per the CLP classification criteria. 

2)Long term toxicity of test material was performed on fish for 21 days according to OECD Guideline 204. 20 test animals per vessele were treated , the test concentation used for the evaluation of toxicity were 0.041, 0.092, 0.204, 0.479, 1.100, 2.546 mg/L (geometric ratio: 2.3). The no effect concentration (NOEC) was observed to be 0.18 mg/l after 21 days of exposure of test material on fish .Effect concentration was indicated based on arithmetic mean values of measurement concentrations, because some analysis results exceeded ±20 % of nominal concentrations in concentration areas.

Based on the above NOEC value it can be concluded that test substance is toxic to fish in the long term and can be classified as aquatic chronic 2 as per CLP

 

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 solution was prepared in aseptic condition. The test substance was prepared by adding 60.75 mg of test substance in 250 ml of BBM to get the final concentration of 243 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 10⁴cells/ml. Care was taken to have a homogeneous solution for the experiment.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 substance  to various nominal test concentration, EC50 calculated from equation and graphically through probit analysis was found to be 30.9134 mg/L and 31.62 mg/L respectively.Based on the EC50, it can be concluded that the chemical was toxic to aqutic algae and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.

Toxicity to microorganism:

Data available for the test chemicals has been reviewed to determine the toxicity of microorganism .The studies are as mentioned below:

1) The effect of test material on Pseudomonas putida was observed for its Oxygen consumption. Pseudomonas putida, strain DSM 50026 were cultured on 25 g/l nutrient broth , 18 g/l Agar poured into slant-agar tubes. The stock solution was prepared with Tween 80 as dispersant and was stirred for 17 hours, resulting in a white suspension. The further dilutions were prepared from the stock solution while stirring. The nominal test concentration used were 156.25, 312.5, 625, 1250, 2500, 10000 mg/l. The effect concentartion EC50 was after 30 min of eposure to test organism Pseudomonas putida was observed to be >10000 mg/l. Based on the above effect concentration it can be considered that test substance is not toxic for the microorganisms.

2) Effect of test material on bio luminescence property of Photobacterium sp. was evaluated after 15 min of exposure of test material. The EC50 value of test material on Photobacterium sp. after 15 mins was observed to be 202 mg/l. Based on the effect concentration it can be concluded that test material has no adverse effect on microorganism.