2-[2-[4-(diethylamino)phenyl]vinyl]-1,3,3-trimethyl-3H-indolium acetate

Endpoint summary

Description of key information

Short term toxicity to fish:

WoE 2: Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test chemical on Zebra fish (Danio rerio) was determined to be < 0.1562 mg/L by observing the mortality. Thus based on LC50, it can be concluded that the chemical was toxic.

WoE 3: Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test chemical on Zebra fish(Danio rerio) was observed to be 0.625 mg/L.

Under the test condition it is observed that the toxicity ranges from 0.625 to <0.1562 mg/l.

Short term toxicity to aquatic invertebrates:

2: Based on nominal concentrations, experimental median effective Concentrations [EC-50 (48 h)] of test chemical on test Daphnia was determine to be 0.5 mg/L.

3: Based on nominal concentrations, experimental median effective Concentrations [EC-50 (48 h)] of test chemical on Daphnia magna was determine to be <0.25 mg/L but >0.125 mg/L.

4: The median effective concentration (EC50) for the test substance on Daphnia magna was determined to be 0.016 mg/L on the basis of mobility inhibition effects in a 48 hour study.

Under the test condition by observing the effects on the mobility of Daphnia magna, it was consider that the chemical was toxic and can be consider to be classified in aquatic acute 1/chronic 1 as per the CLP classification criteria.

Toxicity to algae and cyanobacteria:

WoE 2: Based on the growth inhibition of green alga Pseudokirchneriella subcapitata by the test chemical, the EC50 was determine to be 1.086 mg/l. Based on the EC50, it can be concluded that the chemical was toxic to the growth of algae.

WoE 3: Based on the growth rate inhibition on Desmodesmus subspicatus after the exposure of test chemical for 72 hours, the EC50 was determined to be 0.13 mg/L.

Under the test condition it is observed that the chemical consider to be toxic as cause toxicity on Pseudokirchneriella subcapitata and Desmodesmus subspicatus at concentrations ranges from 1.086 to 0.13 mg/l.

Toxicity to microorganisms:

A screening method based on the measurement of the respiration rate of activated sludge for assessing the possible inhibitory effect of dyestuffs on aerobic waste-water bacteria. The test principle involves measuring the respiration rate of an activated sludge and comparing it with the respiration rate of the same activated sludge under identical conditions, but in the presence of the chemical under test. The test was carried out in activated sludge respiration rate apparatus with constant 20 ± 2°C and pH about 7-8. The test concentration used was 100 mg/l. OECD recommended synthetic sewage was used as feed, while activated sludge was obtained from a sewage works treating predominantly domestic sewage or from a sewage works treating predominantly industrial waste water. The respiration rate of an activated sludge and the respiration rate of activated sludge with test chemical were noted down. In order to calculate the inhibitory effect of a particular chemical at 100 mg/l test concentration its respiration rate is expressed as a percentage of the mean of the two control respiration rates. Thus, IC50 value (concentration for 50% inhibition of respiration rate) for the test chemical on activated sludge (aerobic bacteria) is determined to be10 - 100 mg/1 after 3 hrs of exposure.

Additional information

Short term toxicity to fish:

Data available for the test chemical and structurally and functionally similar read across chemicals extracted using mechanistic approach has been reviewed to determine the toxicity of test chemical on the mortality of fishes. The studies are as mentioned below:

 

The study was designed to assess the toxic effects of the test chemical on the fishes. Test conducted in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test). Zebra fish (Danio rerio) was used as a test organism. 0.1562mg/L, 0.3125mg/L, 0.625mg/L, 1.25mg/L, 2.5mg/L concentrations were prepared from the stock solution. 8 fishes exposed to the test concentration. 8 fishes was added per concentration. Test conducted under the static system for 96 hours. The effect of test chemical was calculated on the basis of mortality of test fish’s Zebra fish (Danio rerio). Observations including (mortality, pH, Temperature, dissolved oxygen content) were recorded in the interval of 24, 48 and 96 hours. Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test chemical on Zebra fish (Danio rerio) was determined to be < 0.1562 mg/L by observing the mortality. Thus based on LC50 it can be concluded that the chemical was toxic and classified as aquatic acute 1/ chronic 1 as per the CLP criteria.                                                                                                                                                                                                              

 

Above study further supported by the second from experimental source. Principle of this study was to determine the toxic effects of the test chemical on the fishes. Test conducted in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test). Zebra fish (Danio rerio) was used as a test organism. The nominal concentration selected for the experiment was 0.625mg/L, 1.25mg/L, 2.5mg/L, 5mg/L, 10mg/L and test fish were exposed to these concentration for 96 hours. 8 fishes was added per concentration. . Test conducted under the static system for 96 hours. The effect of test chemical was calculated on the basis of mortality of test fish’s Zebra fish (Danio rerio). Observations including (mortality, pH, Temperature, dissolved oxygen content) were recorded in the interval of 24, 48 and 96 hours. Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] of test chemical on Zebra fish (Danio rerio) was observed to be 0.625 mg/L. Thus based on the LC50 value, chemical consider to be toxic and classified as aquatic acute 1/chronic 1 as per the CLP classification criteria.           

 

Thus based on the above studies, it is observed that the chemical is toxic and can be consider to be classified in aquatic acute 1/ chronic 1 category as per the CLP classification criteria.

 

Short term toxicity to aquatic invertebrates:

Data available for the test chemical and structurally and functionally similar read across chemicals extracted using mechanistic approach has been reviewed to determine the toxicity of test chemical on the mobility of aquatic invertebrates. The studies are as mentioned below:

 

Principle of this study was to observe the effect of test chemical on the mobility pattern of aquatic invertebrates. Test conducted in accordance with OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna was used as a test organism. The test stock solution was prepared by dissolving 100 mg of the test substance in 100 ml of ADaM’s media, further test concentrations 0.125mg/L, 0.25mg/L, 0.5mg/L, 1mg/L, and 2mg/L respectively was prepared from stock solution. 10 Daphnids was added per concentration. Test conducted under the static system for 48 hours. The effect of test chemical was calculated on the basis of immobility observations of Daphnia magna. Observations including (immobility, pH, Temperature, dissolved oxygen content) were recorded in the interval of 24 and 48 hours. Based on nominal concentrations, experimental median effective Concentrations [EC-50 (48 h)] of test chemical on Daphnia was determine to be 0.5 mg/L. Based on the EC50 value, it can be consider that the chemical was toxic and can be consider to be classified as aquatic acute 1/ chronic 1 as per the CLP classification criteria. 

 

Above study was further supported by the study from experimental source. Principle of this study was to observe the effect of test chemical on the mobility pattern of aquatic invertebrates. Test conducted in accordance with OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Daphnia magna was used as a test organism. The test stock solution was prepared by dissolving 100 mg of the test substance in 100 ml of ADaM’s media, further test concentrations 0.125mg/L, 0.25mg/L, 0.5mg/L, 1mg/L, and 2mg/L respectively was prepared from stock solution. 10 Daphnids was added per concentration. Test conducted under the static system for 48 hours. The effect of test chemical was calculated on the basis of immobility observations of Daphnia magna. Observations including (immobility, pH, Temperature, dissolved oxygen content) were recorded in the interval of 24 and 48 hours. Based on nominal concentrations, experimental median effective Concentrations [EC-50 (48 h)] of test chemical on test daphnia was determine to be <0.25 mg/L but >0.125 mg/L. Thus based on the EC50 value, it can be concluded that the test chemical was toxic and can be consider to be classified as aquatic acute 1/ chronic 1 as per the CLP classification criteria.

 

Similar study for determination of the inhibition of the mobility of Daphnia magna was carried out with the test substance conducted according to OECD Guideline 202. Test performed under the static system. The test substance was tested at the concentrations 0, 0.01, 0.02, 0.04, 0.08, 0.16 and 0.32 mg/L. Effects on immobilisation were observed for 48 hours. EC50 was calculated using nonlinear regression by the software Prism 4.0. The median effective concentration (EC50) for the test substance on Daphnia magna was determined to be 0.016 mg/L on the basis of mobility inhibition effects in a 48 hour study. This value indicates that the substance is likely to be hazardous to aquatic invertebrate Daphnia magna and can be classified as aquatic acute category 1/ chronic 1 as per the CLP criteria.

 

Under the test condition by observing the effects on the mobility of Daphnia magna, it was consider that the chemical was toxic and can be consider to be classified in aquatic acute 1/chronic 1 as per the CLP classification criteria.

Toxicity to algae and cyanobacteria:

Data available for the test chemical and structurally and functionally similar read across chemicals extracted using mechanistic approach has been reviewed to determine the toxicity of test chemical on the growth of aquatic algae. The studies are as mentioned below:

 

The study was designed to access the toxic effects of the test chemical on the green alga Pseudokirchneriella subcapitata. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test). Six test concentrations were 0.3mg/l, 0.39mg/l, 0.507mg/l, 0.659mg/l, 0.856mg/l, 1.112mg/l used and the study conducted under the static system. 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. Based on the growth inhibition of green alga Pseudokirchneriella subcapitata by the test chemical, the EC50 was determine to be 1.086 mg/l. Based on the EC50, it can be concluded that the chemical was toxic to the growth of algae.

 

The above study was further supported by the second study from experimental report. Freshwater algal growth inhibition test was carried out on green algae Desmodesmus subspicatus with the substance. Test conducted under the static system for 72 hours and according to the OECD Guideline 201 (Alga, Growth Inhibition Test). The test substance was prepared in OECD growth medium and tested at the concentrations 0, 3, 6, 12, 24 and 50 mg/L. Effects on the growth rate of the organism

were studied. 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 (EC50) of the test substance on algaewas determined to be 0.13 mg/L on the basis of growth rate inhibition effects in a 72 hour study. Basedon the EC50 value, it is concluded that the substance is likely to be hazardous to aquatic algae and canbe consider to be classified as aquatic acute/ chronic 1 category as per the CLP classification criteria.

Thus based on the experimental data from various sources conducted on two different species of algae, concluded that the test chemical is toxic and can be consider to be classified in aquatic acute category 1/ chronic 1 as per the CLP classification criteria.

 

Toxicity to microorganisms:

Data available for the test chemical has been reviewed to determine the toxicity on microorganisms of the test chemical and structurally and functionally similar read across chemicals. The studies are as mentioned below:

 

A screening method based on the measurement of the respiration rate of activated sludge for assessing the possible inhibitory effect of dyestuffs on aerobic waste-water bacteria. The test principle involves measuring the respiration rate of an activated sludge and comparing it with the respiration rate of the same activated sludge under identical conditions, but in the presence of the chemical under test. The test was carried out in activated sludge respiration rate apparatus with constant 20 ± 2°C and pH about 7-8. The test concentration used was 100 mg/l. OECD recommended synthetic sewage was used as feed, while activated sludge was obtained from a sewage works treating predominantly domestic sewage or from a sewage works treating predominantly industrial waste water. The respiration rate of an activated sludge and the respiration rate of activated sludge with test chemical were noted down. In order to calculate the inhibitory effect of a particular chemical at 100 mg/l test concentration its respiration rate is expressed as a percentage of the mean of the two control respiration rates. Thus, IC50 value (concentration for 50% inhibition of respiration rate) for the test chemical on activated sludge (aerobic bacteria) is determined to be10 - 100 mg/1 after 3 hrs of exposure.

 

Above first study was supported by the second study from peer reviewed journal. A screening method based on the measurement of the respiration rate of activated sludge for assessing the possible inhibitory effect of dyestuffs on aerobic waste-water bacteria were carried out. The test principle involves measuring the respiration rate of an activated sludge and comparing it with the respiration rate of the same activated sludge under identical conditions, but in the presence of the chemical under test. The test was carried out in activated sludge respiration rate apparatus with constant 20 ± 2°C and pH about 7-8. The test concentration used was 100 mg/l. OECD recommended synthetic sewage was used as feed, while activated sludge was obtained from a sewage works treating predominantly domestic sewage or from a sewage works treating predominantly industrial waste water. The respiration rate of an activated sludge and the respiration rate of activated sludge with test chemical were noted down. In order to calculate the inhibitory effect of a particular chemical at 100 mg/l test concentration its respiration rate is expressed as a percentage of the mean of the two control respiration rates. Thus, IC50 value (concentration for 50% inhibition of respiration rate) for the test chemical on activated sludge (aerobic bacteria) is determined to be 10 - 100 mg/1 after 3 hrs of exposure.

 

Based on the available data, it is concluded that the test chemical is likely to be toxic to microorganism in the concentration range of 10-100 mg/l.

 

 

Thus based on the overall results and effects observations on the mortality, immobility and growth rate inhibition of aquatic organisms, it can be concluded that the test chemical is toxic and can be consider to be classified as aquatic acute 1/chronic 1 as per the CLP classification criteria.