Registration Dossier

Ecotoxicological information

Endpoint summary

Administrative data

Description of key information

Short term toxicity to fish:

Study was conducted to investigate the acute toxicity of the test item to Pimephales promelas (Fathead Minnow) was determined in a 96-hour static test according to the standards. Test chemical 40.5 g/l were used and prepared by stirring. Test chemical analytically determined by Gas liquid chromatography. Test performed on 48-50 days old Pimephales promelas. Nominal concentrations 0, 1.88, 3.75, 7.50, 15 and 30 mg/l was used. Based on the mortality of test organism Fathead Minnow by the chemical Nicotine sulfate exposure for 96 hrs, the LC50 was determine to be 15.6 mg/l. Based on the LC50, chemical was consider as toxic and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria. But the chemical was readily biodegradable in water thus before exposure with the test fish, chemical degrade. Thus on that criteria chemical was consider as nontoxic and not classified as per the CLP classification criteria.

Short term toxicity to aquatic invertebrates:

The objective of the present study was to develop the method for determination of toxicity of Nicotine sulfate on the growth of aquatic invertebrate daphnia magna by satisfying the guideline. According to the OECD guideline 202 daphnia magna maintain in the appropriate system and follows all the criteria which meet the guideline. Chemical analyzed with a Hewlett-Packard 5840A gas chromatograph equipped with model 7671 A auto injector and also with FID. 0-24 hrs old daphnia magna obtain from the stock-culture units of the Environmental Research Laboratory-Duluth. Organisms not fed 24 hrs before and during the test. Test conducted in 250 ml beakers with two 1.9 cm holes located 1 cm from the bottom and on opposite sides of each beaker. 20 daphnia per concentration were exposed for 48 hrs and the immobility were observed. Adult daphnids were separated into 2liter battery jars. After 24 hrs, 0-24 hrs old daphnia were carefully collected using fire polished glass tubes and squeeze bulbs, counted into 250 ml exposure beakers which were randomly hung in the 10 × 30 cm compartments of the 12 exposure tanks. After 48 hrs immobility and dead organisms were measured by dissecting microscope. Based on the immobility of daphnia magna by the chemical exposure Nicotine sulfate for 48 hrs, the EC50 was determine to be 3.25 mg/l with the 95 % CI of 2.72-3.88 mg/l. Based on the EC50 chemical was consider as toxic and classified as chronic 2 but as Nicotine sulfate was readily biodegrade when comes in contact with water, thus on that criteria it was consider as nontoxic and not classified as per the CLP classification criteria.

Toxicity to aquatic algae:

Short term toxicity study on aquatic algae was conducted for 96 hrs for test substance 3-[(2S)-1-methylpyrrolidin-2-yl] pyridine; sulfuric acid. Test performed in accordance with OECD guideline 201. Chemical was analytically monitored by HPLC and spectrophotometer. 10,000 mg/l test compound dissolved in sterile ASTM (American Society for Testing and Materials) Type II water and prepared stock solutions with different concentrations 0, 10, 20, 40, 80, and 160 mg/l. 125-ml Erlenmeyer flasks were used, producing a final volume of 50 ml with 1-ml aliquot of a concentrated cell suspension (7.2 × 104cells/ml). Algal growth was measured spectrophotometrically (at 350 nm) at 0, 24, 48, 72, and 96 h using filtered medium as a blank. Correct appearance of the algal cells was verified by microscopic examination before testing. A range-finding test (0, 0.1, 1.0, 10, 100, and 1,000 mg/L) was conducted to determine the median effective concentration (EC50) and the median effective concentrations for inhibition of algal growth (ErC50) and biomass accumulation (EbC50). Thus based on the range finding study definitive concentration were selected. Culture was maintained in algal medium at 24 ± 2 °C with illumination of between 4,500 and 4,710 lux. The average specific growth rate was calculated by dividing the natural log of the change in cells/ml between selected assays by the elapsed time (in h) between the assays. The percentage inhibition in average cell growth rate was calculated as the difference between the control and each chemical test concentration. After the exposure of test substance 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid with freshwater algae Selenastrum capricornutum, effects on biomass and growth rate were observed. Based on the biomass inhibition EC50 was determine to be 115 mg/l and on the basis of growth rate inhibition the EC50 was determine to be 72.9 mg/l. The NOEC was observed at 10 mg/l. As the chemical was readily biodegradable, thus consider as nontoxic and not classified.

Toxicity to microorganisms:

study was conducted to determine the toxicity of test chemical Nicotine sulphate on the growth of microorganisms. Test conducted on two different organisms Ruminococcus albus and Butyrivibrio fibrisolvens. As the two different species Ruminococcus albus and Butyrivibrio fibrisolvens exposed with the test chemical nicotine sulphate, effect on the growth of organisms vary. The IGC for Ruminococcus albus was determine to be 10 mg/l and on the other species it was observed that the growth inhibited at 100 mg/l.

 

Additional information

Summarized result of toxicity of Nicotine sulfate (3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid) (65-30-5) on the growth and other activity of fish, aquatic invertebrates, algae and cyanobacteria and microorganisms by considering the data for target chemical are as follows:  

Short term toxicity to fish:

Various experimental data for the target compound Nicotine sulfate (65-30-5) and supporting studies were reviewed for the toxicity on the fish end point which are summarized as below: 

In the first key study for the target chemical Nicotine sulfate (65-30-5) from peer reviewed journal 1990, study was conducted to investigate the acute toxicity of the test item to Pimephales promelas (Fathead Minnow) was determined in a 96-hour static test according to the standards. Test chemical 40.5 g/l were used and prepared by stirring. Test chemical analytically determined by Gas liquid chromatography. Test performed on 48-50 days old Pimephales promelas. Nominal concentrations 0, 1.88, 3.75, 7.50, 15 and 30 mg/l was used. Based on the mortality of test organism Fathead Minnow by the chemical Nicotine sulfate exposure for 96 hrs, the LC50 was determine to be 15.6 mg/l. Based on the LC50, chemical was consider as toxic and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria. But the chemical was readily biodegradable in water thus before exposure with the test fish, chemical degrade. Thus on that criteria chemical was consider as nontoxic and not classified as per the CLP classification criteria.

 

First study was supported by the second experimental study for the test chemical from peer reviewed journal 1988. Study was conducted to investigate the acute toxicity of the test item to Pimephales promelas was determined in a 96-hour static test according to the standards. Test chemical 100 mg/l were used and prepared by stirring. Test chemical analytically determined by Gas liquid chromatography. Test performed on 29 days old Pimephales promelas. Nominal concentrations 0, 2.96, 4.56, 7.02, 10.8 and 16.6 mg/l was used. In 2 liter tank different concentrations were added with 0.850 g/l Pimephales promelas. After the exposure for 96 hrs affected fish lost schooling behavior, were hyper- active and swam near the tank surface. They were over- reactive to external stimuli, had increased respiration, and darkly colored. Also had spinal deformities and lost equilibrium prior to death. Based on the mortality of test organism by the chemical Nicotine sulfate exposure for 96 hrs, the LC50 was determine to be 12.2 mg/l. Based on the LC50, chemical was concluded as toxic and can be consider to be classified aquatic chronic 3 but as the chemical was readily biodegradable thus consider as nontoxic and not classified as per the CLP classification criteria.

 

Similarly in accordance with another study from journal 1997, study was conducted to develop the procedures that relate modes of acute toxic action in the fathead minnow (Pimephales promelas) to chemical structures and properties. An empirically derived database for diverse chemical structures of acute toxicity and corresponding modes of toxic action was developed through joint toxic action studies, the establishment of toxicodynamic profiles, and behavioral and dose–response interpretation of 96-h LC50 tests. 28 - 30 d old juvenile test organisms were used for the study. The test was performed under the flow-through condition with temp of 25 ± 1°C. Test was conducted using Lake Superior water at 25±1°C. Aqueous toxicant concentrations were measured in tests 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. Tests conducted which were conducted on the Benoit and electronic diluters did not have replicate tank exposures. Median lethal concentration (LC50) was calculated using the Trimmed Spearman–Karber Method, with 95% confidence intervals being calculated when possible. Behavioral signs of stress identified for fathead minnows exposed to reference toxicants. Fish shows type III syndrome which indicates spontaneous locomotor activity, convulsion, spasms, tetany, scoliosis, lordosis, and/or hemorrhaging in the vertebral column. After the exposure of 96 hrs of chemical Nicotine sulfate with the Fathead Minnow (Pimephales promelas), the LC50 was observed to be 13.8 mg/l. Based on the LC50 chemical was consider as toxic and classified as chronic 3 but as Nicotine sulfate was readily biodegrade when comes in contact with water, thus on that criteria it was consider as nontoxic and not classified as per the CLP classification criteria.

 

In the fourth supporting study for the test chemical from experimental peer reviewed journal 1987, supports the classification of test chemical. Study was conducted to investigate the effect of test chemical on the mortality rate of fishes Pimephales promelas and Carassius auratus. According to the OECD guideline 202 test organisms were maintain in the appropriate system and follows all the criteria which meet the guideline. Chemical analyzed with a Hewlett-Packard 5840A gas chromatograph equipped with model 7671 A auto injector and also with FID. Both fishes obtain from the stock-culture units of the Environmental Research Laboratory-Duluth. Organisms not fed 24 hrs before and during the test. Test conducted in glass aquaria tank 60×30×30 cm deep. Tank divided into six compartments and one wide compartment with stainless steel screen divider. One standpipe were also present at the top. Test performed in duplicates with control which simultaneously run with the test chemical. Effect calculated by Trimmed Spearman- Karber method were used. As the test conducted on the two species of fish different effect concentrations were obtain at which 50 % mortality were observed. Based on the mortality of Pimephales promelas (Fathead Minnow) by the exposure of Nicotine sulfate for 96 hrs, the LC50 was determine to be 19.7 mg/l. And based on the mortality of Carassius auratus (Goldfish), the LC50 was determine to be 13.1 mg/l. Based on the LC50 chemical was consider as toxic and classified as chronic 3 but as Nicotine sulfate was readily biodegrade when comes in contact with water, thus on that criteria it was consider as nontoxic and not classified as per the CLP classification criteria.

 

Similarly in the next study from the same source conducted on two different species of fish. Study was conducted to investigate the effect of test chemical Nicotine sulfate on the mortality rate of fishes Oncorhynchus mykiss (Rainbow Trout) and Lepomis macrochirus (Bluegill) by providing the exposure period of 96hrs. According to the OECD guideline 202 test organisms were maintain in the appropriate system and follows all the criteria which meet the guideline. Chemical analyzed with a Hewlett-Packard 5840 A gas chromatograph equipped with model 7671 A auto injector and also with FID. Both fishes obtain from the stock-culture units of the Environmental Research Laboratory-Duluth. Organisms not fed 24 hrs before and during the test. Test conducted in glass aquaria tank 60×30×30 cm deep. Tank divided into six compartments and one wide compartment with stainless steel screen divider. One standpipe were also present at the top. Test performed in duplicates with control which simultaneously run with the test chemical. Effect calculated by Trimmed Spearman- Karber method were used. As the test conducted on the two species of fish different effect concentrations were obtain at which 50 % mortality were observed. Based on the mortality of Oncorhynchus mykiss (Rainbow Trout) by the exposure of Nicotine sulfate for 96 hrs, the LC50 was determine to be 7.31 mg/l. And based on the mortality of Lepomis macrochirus (Bluegill), the LC50 was determine to be 4.31 mg/l. Based on the LC50 chemical was consider as toxic and classified as chronic 2 but as Nicotine sulfate was readily biodegrade when comes in contact with water, thus on that criteria it was consider as nontoxic and not classified as per the CLP classification criteria.

 

Based on the overall experimental studies it was observed that the chemical was slightly toxic but as when comes in contact with water it readily degrade and thus on all criteria, it can be concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.

 

Short term toxicity to aquatic invertebrates:

In the first key study from the peer reviewed journal 1987 toxicity on invertebrates were studied. The objective of the present study was to develop the method for determination of toxicity of Nicotine sulfate on the growth of aquatic invertebrate daphnia magna by satisfying the guideline. According to the OECD guideline 202 daphnia magna maintain in the appropriate system and follows all the criteria which meet the guideline. Chemical analyzed with a Hewlett-Packard 5840A gas chromatograph equipped with model 7671 A auto injector and also with FID. 0-24 hrs old daphnia magna obtain from the stock-culture units of the Environmental Research Laboratory-Duluth. Organisms not fed 24 hrs before and during the test. Test conducted in 250 ml beakers with two 1.9 cm holes located 1 cm from the bottom and on opposite sides of each beaker. 20 daphnia per concentration were exposed for 48 hrs and the immobility were observed. Adult daphnids were separated into 2liter battery jars. After 24 hrs, 0-24 hrs old daphnia were carefully collected using fire polished glass tubes and squeeze bulbs, counted into 250 ml exposure beakers which were randomly hung in the 10 × 30 cm compartments of the 12 exposure tanks. After 48 hrs immobility and dead organisms were measured by dissecting microscope. Based on the immobility of daphnia magna by the chemical exposure Nicotine sulfate for 48 hrs, the EC50 was determine to be 3.25 mg/l with the 95 % CI of 2.72-3.88 mg/l. Based on the EC50 chemical was consider as toxic and classified as chronic 2 but as Nicotine sulfate was readily biodegrade when comes in contact with water, thus on that criteria it was consider as nontoxic and not classified as per the CLP classification criteria.

 

First study was supported by the second study from the peer reviewed journal 1987. The objective of the present study was to develop the method for determine the toxicity of Nicotine sulfate on the mortality of Aplexa hypnorum (Snail). According to the OECD guideline 202 Aplexa hypnorum (Snail) maintain in the appropriate system and follows all the criteria which meet the guideline. Chemical analyzed with a Hewlett-Packard 5840A gas chromatograph equipped with model 7671 A auto injector and also with FID. Adult Aplexa hypnorum (Snail) obtain from the stock-culture units of the Environmental Research Laboratory-Duluth. Organisms not fed 24 hrs before and during the test. Test conducted in glass aquaria tank 60×30×30 cm deep. Tank divided into six compartments and one wide compartment with stainless steel screen divider. One standpipe were also present at the top. After 96 hrs dead organisms were measured by dissecting microscope. Based on the mortality of Aplexa hypnorum snail by the chemical exposure Nicotine sulfate for 96 hrs, the EC50 was determine to be > 38.2 mg/l. Based on the EC50 chemical was consider as toxic and classified as chronic 3 but as Nicotine sulfate was readily biodegrade when comes in contact with water, thus on that criteria it was consider as nontoxic and not classified as per the CLP classification criteria.

 

Based on the above studies it can be concluded that the chemical was nontoxic and not classified as per the CLP classification criteria.

 

Toxicity to aquatic algae:

In the key study from the peer reviewed journal 2009, Short term toxicity study on aquatic algae was conducted for 96 hrs for test substance 3-[(2S)-1-methylpyrrolidin-2-yl] pyridine; sulfuric acid. Test performed in accordance with OECD guideline 201. Chemical was analytically monitored by HPLC and spectrophotometer. 10,000 mg/l test compound dissolved in sterile ASTM (American Society for Testing and Materials) Type II water and prepared stock solutions with different concentrations 0, 10, 20, 40, 80, and 160 mg/l. 125-ml Erlenmeyer flasks were used, producing a final volume of 50 ml with 1-ml aliquot of a concentrated cell suspension (7.2 × 104cells/ml). Algal growth was measured spectrophotometrically (at 350 nm) at 0, 24, 48, 72, and 96 h using filtered medium as a blank. Correct appearance of the algal cells was verified by microscopic examination before testing. A range-finding test (0, 0.1, 1.0, 10, 100, and 1,000 mg/L) was conducted to determine the median effective concentration (EC50) and the median effective concentrations for inhibition of algal growth (ErC50) and biomass accumulation (EbC50). Thus based on the range finding study definitive concentration were selected. Culture was maintained in algal medium at 24 ± 2 °C with illumination of between 4,500 and 4,710 lux. The average specific growth rate was calculated by dividing the natural log of the change in cells/ml between selected assays by the elapsed time (in h) between the assays. The percentage inhibition in average cell growth rate was calculated as the difference between the control and each chemical test concentration. After the exposure of test substance 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid with freshwater algae Selenastrum capricornutum, effects on biomass and growth rate were observed. Based on the biomass inhibition EC50 was determine to be 115 mg/l and on the basis of growth rate inhibition the EC50 was determine to be 72.9 mg/l. The NOEC was observed at 10 mg/l. As the chemical was readily biodegradable, thus consider as nontoxic and not classified.

 

Toxicity to microorganisms:

In the study from peer reviewed journal 1977, study was conducted to determine the toxicity of test chemical Nicotine sulphate on the growth of microorganisms. Test conducted on two different organisms Ruminococcus albus and Butyrivibrio fibrisolvens. As the two different species Ruminococcus albus and Butyrivibrio fibrisolvens exposed with the test chemical nicotine sulphate, effect on the growth of organisms vary. The IGC for Ruminococcus albus was determine to be 10 mg/l and on the other species it was observed that the growth inhibited at 100 mg/l.

 

 

Based on the overall experimental studies it was observed that the chemical was slightly toxic but as when comes in contact with water it readily degrade and thus on all criteria, it can be concluded that the chemical was nontoxic to aquatic life and not classified as per the CLP classification criteria.