N-acetylsulphanilyl chloride

Endpoint summary

Description of key information

Short term toxicity to fish

Short term toxicity to fish was conducted for 96 hrs for assessing the effect of test chemical (Experimental study report, 2013). The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.203 g and average length of 2.3 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.4 mg/l, pH 7 to 8, water temperature 29°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. The limit test (100 mg/l) was conducted for this substance showed 100% mortalities within 24 hours of the start of the experiment. Thus, on the basis of this, definitive test was performed at different test chemical nominal concentrations, i.e., at 0, 0.9, 2, 5, 15, 25, 50, 75 and 100 mg/l (nominal concentrations), respectively. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 29°C, pH 7.0, hardness of water 155.5 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control vessel. Test fishes were found to be freely swimming in the bowl aquaria but mortalities were observed in the test vessels during the study. On the basis of effect of test chemical on mortality of the test organism, the LC0, median lethal concentration (LC50 (96 h)) and LC100 value was determined to be 2 mg/l, > 2 to < 5 mg/L and 5 mg/l, respectively. Thus, based on the LC50 value, test chemical can be considered as toxic to aquatic fishes. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic fishes at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

Long term toxicity to fish

The long-term toxicity of the test chemical to aquatic fish was predicted using EPI Suite ECOSAR version 1.11. On the basis of effect of test chemical observed in a flow through system on the reproduction of the test organism during the 28 days exposure duration, the NOEC value for the test chemical was estimated to be 17.079 mg/l. Thus, based on the NOEC value, test chemical was considered as non-toxic to aquatic fishes and hence, considered to be 'not classified' as per the CLP classification criteria.

Short term toxicity to aquatic invertebrates

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates (Experimental study report, 2018). The test was performed following the principles of the OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The stock solution 50 mg/l was prepared by dissolving test chemical in reconstituted water. The solution was kept in ultrasonic bath for 20 min. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water.Test chemical concentrations were not verified analytically. Nominal test chemical concentrations used for the study were 0, 10, 18, 32, 58 and 100 mg/l, respectively. Study was performed using 5 organisms per vessel/replicates in a static fresh water system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. EC50 was calculated using non linear regression by the software Prism 4. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test. The 24 hr EC50 value of the reference substance was determined to be 0.73 mg/l. The dissolved oxygen concentration at the end of the test both in the control and test vessels was evaluated to be ≥ 3 mg/l (i.e, reported as > 8.2 mg/l), indicating that the validity criteria has been fulfilled. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be 48.6 mg/l (95 % C. I. - 36.1 to 65.5 mg/l) (nominal concentration). Thus, test chemical was considered as toxic to aquatic invertebrates. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

Long term toxicity to aquatic invertebrates

The long-term toxicity of the test chemical to aquatic invertebrates was predicted using EPI Suite ECOSAR version 1.11. On the basis of effect of test chemical observed in a flow through system on the reproduction of the test organism during the 21 days exposure duration, the NOEC value for the test chemical was estimated to be 9.788 mg/l. Thus, based on the NOEC value, test chemical was considered as non-toxic to aquatic invertebrates and hence, considered to be 'not classified' as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria

Toxicity to aquatic algae study was conducted for 72 hrs for assessing the effect of test chemical on green algae (Experimental study report, 2018). The test was performed in accordance to OECD Guideline 201 (Alga, Growth Inhibition Test) in a static system. Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) of strain 86.81 SAG obtained from Institute of botany of the ASCR with an initial biomass conc. 5000 cells /ml was used as a test organism. The stock solution (100 mg/L) was prepared by dissolving test chemical in OECD growth medium. The stock solution was kept in ultrasonic bath for 20 mins. Test solutions  of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture.Test chemical concentrations were not verified analytically. Nominal test chemical conc. used for the study were 0, 10, 17, 29, 49 and 84 mg/l, respectively. Desmodesmus subspicatus were exposed to test chemical in 50 ml glass vessel in a volume of 15 ml of liquid solution containing both the chemical and media. Control solution vessel containing OECD medium without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 23±2°C with a continuous light intensity of 6000-8000 lx, respectively. Alongwith the test chemical, one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. Cell counting was carried out using microscope with counting chamber Cyrus I or electronic particle counter. ErC50 was calculated using non-linear regression by the software Prism 4.0. In the control test vessel containing OECD growth medium without test chemical, the coefficient of variation of average growth rate in replicates during the whole test period was 2.8% i.e, not exceeded 7% and the mean coefficient of variation for section by section specific growth rate in the control cultures was not exceeded 35%, indicating that the validity criteria has been fulfilled. On the basis of the effect of test chemical on the growth rate of the test organism Desmodesmus subspicatus, the 72 hr median effect concentration (ErC50) value was determined to be 70.8 mg/l (95 % CI 65.2 - 76.8 mg/l) (nominal concentration). Thus, based on the EC50 value, test chemical can be considered as toxic to aquatic algae. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic algae at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

Toxicity to microoganisms

On the basis of the various experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism, the 3 hr EC50 value can be expected to be > 500 mg/l.

Additional information

Short term toxicity to fish

Experimental study and various predicted data of the target chemical and supporting weight of evidence study of its structurally similar read across substance were reviewed for the short term toxicity to aquatic invertebrates end point which are summarized as below:

 

In an experimental key study from study report (2013),short term toxicity to fish was conducted for 96 hrs for assessing the effect of test chemical. The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.203 g and average length of 2.3 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.4 mg/l, pH 7 to 8, water temperature 29°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. The limit test (100 mg/l) was conducted for this substance showed 100% mortalities within 24 hours of the start of the experiment. Thus, on the basis of this, definitive test was performed at different test chemical nominal concentrations, i.e., at 0, 0.9, 2, 5, 15, 25, 50, 75 and 100 mg/l (nominal concentrations), respectively. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 29°C, pH 7.0, hardness of water 155.5 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control vessel. Test fishes were found to be freely swimming in the bowl aquaria but mortalities were observed in the test vessels during the study. On the basis of effect of test chemical on mortality of the test organism, the LC0, median lethal concentration (LC50 (96 h)) and LC100 value was determined to be 2 mg/l, > 2 to < 5 mg/L and 5 mg/l, respectively. Thus, based on the LC50 value, test chemical can be considered as toxic to aquatic fishes. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic fishes at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria..

 

In a prediction done using EPI Suite ECOSAR version 1.11, theshort-term toxicity of the test chemical to aquatic fish was predicted.On the basis of effect of test chemical observed in a static system on the mortality of the test organism during the 96 hr exposure duration, the lethal effect concentration (LC50) for the test chemical was estimated to be 173.818 mg/l. Thus, based on the LC50 value, test chemical can be considered as non-toxic to aquatic fishes and hence, considered to be ‘not classified’ as per CLP classification criteria.

 

In an another prediction done using Danish QSAR database, the 96 hrs LC50 value of test chemical on the test organism Pimephales promelas (Fathead minnow) was estimated by three different models i.e, Battery, Leadscope and SciQSAR used within Danish QSAR database. On the basis of effect of test chemical on the mortality of the test organism, the 96 hrs LC50 value was estimated to be 61.39 mg/l. Thus, based on the LC50 value, chemical can be considered as toxic to aquatic fishes. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic and hence, considered to be 'not classified' as per the CLP classification criteria.

 

For the test chemical, short term fish toxicity was conducted for 96 hrs for assessing the effect of test chemical (authoritative database, 2020). Study was carried out under static conditions. On the basis of the effect of test chemical on mortality of the test fishes, the 96 hrs median lethal concentration (LC50) value was determined to be 77 mg/l. Thus, based on the LC50 value, test chemical can be considered as toxic to aquatic fishes. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic fishes at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

 

On the basis of the above results, it can be concluded that the test chemical was considered as toxic to aquatic fish. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic fishes at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

Long term toxicity to fish

The long-term toxicity of the test chemical to aquatic fish was predicted using EPI Suite ECOSAR version 1.11. On the basis of effect of test chemical observed in a flow through system on the reproduction of the test organism during the 28 days exposure duration, the NOEC value for the test chemical was estimated to be 17.079 mg/l. Thus, based on the NOEC value, test chemical was considered as non-toxic to aquatic fishes and hence, considered to be 'not classified' as per the CLP classification criteria.

Short term toxicity to aquatic invertebrates

Experimental study & predicted data of the target chemical and supporting weight of evidence study of its structurally similar read across substance were reviewed for the short term toxicity to aquatic invertebrates end point which are summarized as below:

 

In an experimental key study from study report (2018),an acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates. The test was performed following the principles of the OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The stock solution 50 mg/l was prepared by dissolving test chemical in reconstituted water. The solution was kept in ultrasonic bath for 20 min. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water. Test chemical concentrations were not verified analytically. Nominal test chemical concentrations used for the study were 0, 10, 18, 32, 58 and 100 mg/l, respectively. Study was performed using 5 organisms per vessel/replicates in a static fresh water system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. EC50 was calculated using non-linear regression by the software Prism 4. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test. The 24 hr EC50 value of the reference substance was determined to be 0.73 mg/l. The dissolved oxygen concentration at the end of the test both in the control and test vessels was evaluated to be ≥ 3 mg/l (i.e, reported as > 8.2 mg/l), indicating that the validity criteria has been fulfilled. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be 48.6 mg/l (95 % C. I. - 36.1 to 65.5 mg/l) (nominal concentration). Thus, test chemical was considered as toxic to aquatic invertebrates. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

 

In a prediction done using EPI Suite ECOSAR version 1.11, theshort-term toxicity of the test chemical to aquatic invertebrates was predicted. On the basis of effect of test chemical observed in a static system on the mobility of the test organism during the 48 hr exposure duration, the lethal effect concentration (LC50) for the test chemical was estimated to be 99.133 mg/l. Thus, based on the LC50 value, test chemical can be considered as toxic to aquatic invertebrates. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic and hence, considered to be 'not classified' as per the CLP classification criteria.

 

For the test chemical from authoritative database (2020), an acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical. The test was performed following the principles of the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The study was performed under static conditions using Daphnia magna (Water flea) as a test organism. On the basis of the toxic effect of the test chemical on mobility of the test organism Daphnia magna, the 48 hr median effect concentration (EC50) value was determined to be 72 mg/l (nominal conc.). Thus, test chemical was considered as toxic to aquatic invertebrates. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

 

On the basis of the above results, it can be concluded that the test chemical was considered as toxic to aquatic invertebrates. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

Long term toxicity to aquatic invertebrates

The long-term toxicity of the test chemical to aquatic invertebrates was predicted using EPI Suite ECOSAR version 1.11. On the basis of effect of test chemical observed in a flow through system on the reproduction of the test organism during the 21 days exposure duration, the NOEC value for the test chemical was estimated to be 9.788 mg/l. Thus, based on the NOEC value, test chemical was considered as non-toxic to aquatic invertebrates and hence, considered to be 'not classified' as per the CLP classification criteria.

Toxicity to aquatic algae and cyanobacteria

Various experimental studies and predicted data of the target chemical and supporting weight of evidence study of its structurally similar read across substance were reviewed for the toxicity to aquatic algae and cyanobacteria end point which are summarized as below:

 

In an experimental key study from study report (2018), toxicity to aquatic algae study was conducted for 72 hrs for assessing the effect of test chemical on green algae. The test was performed in accordance to OECD Guideline 201 (Alga, Growth Inhibition Test) in a static system. Desmodesmus subspicatus (previous name: Scenedesmus subspicatus) of strain 86.81 SAG obtained from Institute of botany of the ASCR with an initial biomass conc. 5000 cells /ml was used as a test organism. The stock solution (100 mg/L) was prepared by dissolving test chemical in OECD growth medium. The stock solution was kept in ultrasonic bath for 20 mins. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. Test chemical concentrations were not verified analytically. Nominal test chemical conc. used for the study were 0, 10, 17, 29, 49 and 84 mg/l, respectively. Desmodesmus subspicatus were exposed to test chemical in 50 ml glass vessel in a volume of 15 ml of liquid solution containing both the chemical and media. Control solution vessel containing OECD medium without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 23±2°C with a continuous light intensity of 6000-8000 lx, respectively. Alongwith the test chemical, one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. Cell counting was carried out using microscope with counting chamber Cyrus I or electronic particle counter. ErC50 was calculated using non-linear regression by the software Prism 4.0. In the control test vessel containing OECD growth medium without test chemical, the coefficient of variation of average growth rate in replicates during the whole test period was 2.8% i.e, not exceeded 7% and the mean coefficient of variation for section by section specific growth rate in the control cultures was not exceeded 35%, indicating that the validity criteria has been fulfilled. On the basis of the effect of test chemical on the growth rate of the test organism Desmodesmus subspicatus, the 72 hr median effect concentration (ErC50) value was determined to be 70.8 mg/l (95 % CI 65.2 - 76.8 mg/l) (nominal concentration). Thus, based on the EC50 value, test chemical can be considered as toxic to aquatic algae. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic algae at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

 

Another freshwater algal growth inhibition test was conducted for 72 hrs for assessing the effect of test chemical on green algae Chlorella vulgaris (Experimental study report, 2014). The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. Bold’s Basal Medium (BBM) composed of macronutrients, micronutrients, alkaline EDTA solution and iron solution was used as a growth medium. The test substance was prepared by dissolving 60 mg of test substance in 300 ml of BBM to get the final concentration of 200 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 10000 cells/ml. Green algae were exposed to nominal concentration of test chemical (0, 6.25, 12.5, 25, 50, 100 and 200 mg/l) in 100 ml conical flasks. Test vessel were placed in orbital shaking incubator for 72 hrs at a room at a temperature of 24±2°C under a photoperiod of 16:8 hr light: dark conditions and with a continuous uniform illumination of 3000-4000 lux light intensity, respectively. The speed of the orbital shaking incubator was set at a 120 revolutions per minute throughout the study period. Control containing medium without test chemical was used for the study. The cultures were counted and observed daily with the help of an automated cell counter to verify a normal and healthy appearance of the algae cells and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test chemical). All the cells appeared healthy, round and green throughout the study duration in the control while following changes were observed at the concentration of 200 mg/l which includes, decrease in cell count, discoloration of algal cells, deformed and shrinked cell size and fine particles observed inside the cell. On the basis of growth rate of the test organism Chlorella vulgaris, the 72 hrs median effect concentration (ErC50) value calculated from equation and graphically through probit analysis was determined to be 193.43 and 200 mg/l. On the basis of the EC50 value, chemical was considered to be non-toxic to aquatic algae and hence, considered to be 'not classified' as per the CLP classification criteria.

 

In a prediction done using EPI Suite ECOSAR version 1.11, the toxicity of the test chemical to green algae was predicted. On the basis of effect of test chemical observed in a static system on the growth rate of the test organism during the 96 hr exposure duration, the median effect concentration (EC50) for the test chemical was estimated to be 75.211 mg/l. Thus, based on the EC50 value, test chemical can be considered as toxic to aquatic algae. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic and hence, considered to be 'not classified' as per the CLP classification criteria.

 

For the test chemical, algal toxicity study was carried out for 72 hrs for assessing the effect of test chemical (authoritative database, 2020). The study was performed in accordance with the OECD Guideline 201 (Alga, Growth Inhibition Test). Study was carried out under static conditions. Based on the effect of test chemical on growth rate of the test algae, the 72 hrs NOEC and EC50 value was determined to be 26 mg/l & 57 mg/l, respectively and on the basis of AUG, the 72 hrs NOEC and EC50 value was determined to be 24 mg/l & 41 mg/l, respectively. Thus, based on the EC50 value, test chemical can be considered as toxic to aquatic algae. Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic algae at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

 

On the basis of the above results, it can be concluded that the test chemical was considered as toxic to aquatic algae.Since the test chemical is readily biodegradable in water, test chemical was considered as non-toxic to aquatic algae at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.

Toxicity to microoganisms

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

 

Toxicity study to activated sludge was carried out for 3 hrs for assessing the effect of test chemical. The study was performed in accordance with the OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test. Activated sludge, predominantly domestic sewage was used as a test organism. On the basis of the effect on respiration rate of the test inoculum, the 3 hr EC0 and EC50 value was determined to be 500 and >500 mg/l, respectively.

 

Another toxicity study to activated sludge was carried out for 3 hrs for assessing the effect of test chemical. The study was performed following the principles of the OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test. Activated sludge, predominantly domestic sewage was usedas a test organism. Test chemical concentrations were not analytically verified. On the basis of the effect on respiration rate of the test inoculum, the 3 hr EC0 value was determined to be >1000 mg/l (nominal concentrations).

 

For the test chemical, toxicity study to anaerobic bacteria was carried out for 24 hrs for assessing the effect of test chemical. On the basis of the effect on growth inhibition of the test bacteria, the 24 hr EC10 value was determined to be >5000 mg/l.

 

On the basis of the various experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism, the 3 hr EC50 value can be expected to be > 500 mg/l.

On the basis of the available information of aquatic toxicity studies, it can be concluded that the test chemical was considered as non-toxic at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.