N-(2-chloroethyl)pyrrolidine hydrochloride

Endpoint summary

Description of key information

Short term toxicity to fish

Various experimental data were reviewed for to assess the toxic effects of test compound which are summarized as below:

 

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 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring of 1 hour for achieving test concentrations of 100 mg/L, respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. 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 of test chemical at of 100mg/L, no mortality was observed. Therefore the LC50 was determined to be >100mg/L.

 

The above result was supported by another study. Ninety-six-hour flow-through exposures using 31-d-old juvenile fathead minnows were conducted on 617 chemicals including our target chemical. Tests were conducted using Lake Superior water at 25 (+-) 18C. Aqueous toxicant concentrations were measured in test with quality assurance criteria requiring 80% agreement between duplicate samples and 90 to 110% spike recovery. Flow-through exposures were conducted using cycling proportional, modified Benoit or electronic diluters.

Median lethal concentrations (LC50s) were calculated using the Trimmed Spearman–Karber Method, with 95% confidence intervals being calculated when possible.

Results show 50% mortality after 96 hours at dose concentration 153 mg/l. Therefore, LC50 was considered to be 153 mg/L when exposed to test chemical.

 

Based on the LC50 values, the test chemical can be considered as non-toxic and can be concluded as not classified as per the CLP classification criteria.

 

 

Short-term toxicity to aquatic invertebrates

Determination of the inhibition of the mobility of daphnids was carried out with the substance according to OECD Guideline 202.

The stock solution 100 mg/l was prepared by dissolving white powder in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water.

The test substance was tested at the concentrations 0, 1, 2, 4, 8 and 16 mg/L. Effects on immobilisation were observed for 48 hours.

Effects on immobilisation were observed for 48 hours. 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.

 

The median effective concentration (EC50) for the test substance in Daphnia magna was determined to be 3 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be hazardous to aquatic invertebrate and can be as aquatic chronic 2 category as classified as per the CLP classification criteria.

 

 

 

Toxicity to aquatic algae and cyanobacteria

Various experimental data for the target compound were reviewed for to assess the effects of test chemical on the growth of fresh water green alga along with the predicted data which are summarized as below:

 

Aim of this study was to evaluate the nature of chemical test chemical when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD guideline 201. The stock solution 100 mg/l was prepared by dissolving white powder in OECD growth medium. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. 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 (ErC50) for the test substance in algae was determined to be 4.29 mg/L on the basis of growth rate inhibition effects in a 72 hour study.

 

The above result was supported by another study 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). The test solution was prepared in aseptic condition. The test item N-(2-chloroethyl)pyrrolidine hydrochloride was prepared by adding 54.67 mg of test item in 250 ml of BBM to get the final concentration of 218.7 mg/L. This stock solution was kept for stirring for 30 minutes to obtain a homogenous solution for the experiment. 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 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 0.264 per day.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 20.24%.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 9.31%. Hence, the test is considered valid as per OECD guideline, 201

After 72 hours of exposure to test item N-(2-chloroethyl)pyrrolidine hydrochloride(CAS No. 7250- 67-1) to various nominal test concentrations, EC50 was found to be 11.548 mg/l graphically and through probit analysis.

Based on the EC50 value, which indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 2 category as per the CLP classification criteria.

Toxicity to microorganisms

Three studies which includes reliable predicted data and experimental results from authorative database for micro organism toxicity of test chemical 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) with relevant read across which is structurally similar to target were summarized as follows:

 

First prediction done using QSAR toolbox version.3.3 which indicate the toxicity to micro organism of the substance 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) to micro organism Tetrahymena pyriformis is predicted based on the effects observed in a static freshwater system during a 48 hr exposure. The Inhibition growth concentration (IGC50) for the substance 1-(2-chloroethyl)pyrrolidine hydrochloride is estimated to be 214.11 mg/L. Based on this value, it can be concluded that the test chemical 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) may have no concern to micro organism toxicity for acute exposure period.

 

Above result assist by experimental data of read across 4-methylpent-3-en-2-one (Cas no. 141-79-7) suggest Inhibition concentration to 50% of test organisms (IGC50) for the substance 4-methylpent-3-en-2-one (CAS no. 141-79-7) was determine to be 4.403 mM which is equivalent to 431 mg/l on micro organism species Tetrahymena pyriformis (Ciliate) on the basis of effect on Population growth rate in static fresh water.(ECOTOX DATABASE;2017)

 

And another read across chemical 3-Hexyn-2-one (Cas no. 1679-36-3) from same data source i.e ECOTOX database; 2017 indicate Inhibition concentration to 50% of test organisms (IGC50) for the substance 3-Hexyn-2-one (CAS no. 1679-36-3) was determine to be 0.055 mM which is equivalent to 5.2 mg/l on micro organism species Tetrahymena pyriformis (Ciliate) on the basis of effect on Population growth rate in static fresh water.

 

All available studies give the Inhibition concentration to 50% of test organisms (IGC50) is in the range of 5.2-431 mg/l which indicate the test chemical 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) may have no concern to micro organism toxicity for acute exposure period.

Additional information

Short term toxicity to fish

Various experimental data were reviewed for to assess the toxic effects of test compound which are summarized as below:

 

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 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring of 1 hour for achieving test concentrations of 100 mg/L, respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. 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 of test chemical at of 100mg/L, no mortality was observed. Therefore the LC50 was determined to be >100mg/L.

 

The above result was supported by another study. Ninety-six-hour flow-through exposures using 31-d-old juvenile fathead minnows were conducted on 617 chemicals including our target chemical. Tests were conducted using Lake Superior water at 25 (+-) 18C. Aqueous toxicant concentrations were measured in test with quality assurance criteria requiring 80% agreement between duplicate samples and 90 to 110% spike recovery. Flow-through exposures were conducted using cycling proportional, modified Benoit or electronic diluters.

Median lethal concentrations (LC50s) were calculated using the Trimmed Spearman–Karber Method, with 95% confidence intervals being calculated when possible.

Results show 50% mortality after 96 hours at dose concentration 153 mg/l. Therefore, LC50 was considered to be 153 mg/L when exposed to test chemical.

 

Based on the LC50 values, the test chemical can be considered as non-toxic and can be concluded as not classified as per the CLP classification criteria.

 

 

Short-term toxicity to aquatic invertebrates

Determination of the inhibition of the mobility of daphnids was carried out with the substance according to OECD Guideline 202.

The stock solution 100 mg/l was prepared by dissolving white powder in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with reconstituted test water.

The test substance was tested at the concentrations 0, 1, 2, 4, 8 and 16 mg/L. Effects on immobilisation were observed for 48 hours.

Effects on immobilisation were observed for 48 hours. 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.

 

The median effective concentration (EC50) for the test substance in Daphnia magna was determined to be 3 mg/L on the basis of mobility inhibition effects in a 48 hour study. Based on the EC50 value, substance is likely to be hazardous to aquatic invertebrate and can be as aquatic chronic 2 category as classified as per the CLP classification criteria.

 

 

 

Toxicity to aquatic algae and cyanobacteria

Various experimental data for the target compound were reviewed for to assess the effects of test chemical on the growth of fresh water green alga along with the predicted data which are summarized as below:

 

Aim of this study was to evaluate the nature of chemical test chemical when comes in contact with the test organism Desmodesmus subspicatus (previous name: Scenedesmus subspicatus). Test was conducted according to the OECD guideline 201. The stock solution 100 mg/l was prepared by dissolving white powder in OECD growth medium. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture. 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 (ErC50) for the test substance in algae was determined to be 4.29 mg/L on the basis of growth rate inhibition effects in a 72 hour study.

 

The above result was supported by another study 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). The test solution was prepared in aseptic condition. The test item N-(2-chloroethyl)pyrrolidine hydrochloride was prepared by adding 54.67 mg of test item in 250 ml of BBM to get the final concentration of 218.7 mg/L. This stock solution was kept for stirring for 30 minutes to obtain a homogenous solution for the experiment. 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 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 0.264 per day.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 20.24%.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 9.31%. Hence, the test is considered valid as per OECD guideline, 201

After 72 hours of exposure to test item N-(2-chloroethyl)pyrrolidine hydrochloride(CAS No. 7250- 67-1) to various nominal test concentrations, EC50 was found to be 11.548 mg/l graphically and through probit analysis.

Based on the EC50 value, which indicates that the substance is likely to be hazardous to aquatic algae and can be classified as aquatic chronic 2 category as per the CLP classification criteria.

Toxicity to microorganisms

Three studies which includes reliable predicted data and experimental results from authorative database for micro organism toxicity of test chemical 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) with relevant read across which is structurally similar to target were summarized as follows:

 

First prediction done using QSAR toolbox version.3.3 which indicate the toxicity to micro organism of the substance 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) to micro organism Tetrahymena pyriformis is predicted based on the effects observed in a static freshwater system during a 48 hr exposure. The Inhibition growth concentration (IGC50) for the substance 1-(2-chloroethyl)pyrrolidine hydrochloride is estimated to be 214.11 mg/L. Based on this value, it can be concluded that the test chemical 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) may have no concern to micro organism toxicity for acute exposure period.

 

Above result assist by experimental data of read across 4-methylpent-3-en-2-one (Cas no. 141-79-7) suggest Inhibition concentration to 50% of test organisms (IGC50) for the substance 4-methylpent-3-en-2-one (CAS no. 141-79-7) was determine to be 4.403 mM which is equivalent to 431 mg/l on micro organism species Tetrahymena pyriformis (Ciliate) on the basis of effect on Population growth rate in static fresh water.(ECOTOX DATABASE;2017)

 

And another read across chemical 3-Hexyn-2-one (Cas no. 1679-36-3) from same data source i.e ECOTOX database; 2017 indicate Inhibition concentration to 50% of test organisms (IGC50) for the substance 3-Hexyn-2-one (CAS no. 1679-36-3) was determine to be 0.055 mM which is equivalent to 5.2 mg/l on micro organism species Tetrahymena pyriformis (Ciliate) on the basis of effect on Population growth rate in static fresh water.

 

All available studies give the Inhibition concentration to 50% of test organisms (IGC50) is in the range of 5.2-431 mg/l which indicate the test chemical 1-(2-chloroethyl)pyrrolidine hydrochloride (Cas no. 7250-67-1) may have no concern to micro organism toxicity for acute exposure period.

Based on the overall studies including toxicity to fish, invertebrates, algae and cyanobacteria, it was concluded that the chemical was toxic. Also the test substance is not readily biodegradable and thus can be classified in the chronic 2 classification category as per the CLP classification criteria.