Registration Dossier

Environmental fate & pathways

Biodegradation in soil

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
biodegradation in soil, other
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed journal
Justification for type of information:
Data is from peer reviewed journal
Reference:
Composition 0
Qualifier:
according to
Guideline:
other: Modified OECD Guideline 304 A (Inherent Biodegradability in Soil)
Principles of method if other than guideline:
Biodegradation study in soil was conducted for 28 days for determining the half-life value of the chemical 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid. The study was performed using the Modified OECD Guideline 304 A (Inherent Biodegradability in Soil).
GLP compliance:
not specified
Test type:
not specified
Test material information:
Composition 1
Specific details on test material used for the study:
- Name of test material (IUPAC Name): 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid
- Common name: Nicotine sulphate
- Molecular formula: C20H30N4O4S
- Molecular weight: 422.54 g/mol
- Smiles notation: CN1CCCC1C2=CN=CC=C2.CN1CCCC1C2=CN=CC=C2.OS(=O)(=O)O
- InChI: 1S/2C10H14N2.H2O4S/c2*1-12-7-3-5-10(12)9-4-2-6-11-8-9;1-5(2,3)4/h2*2,4,6,8,10H,3,5,7H2,1H3;(H2,1,2,3,4)/t2*10-;/m00./s1
- Substance type: Organic
- Physical state: Solid
- Locations of the label (if radiolabelling): Radiolabeling was done at the carbon position. The test substance, radiolabeled (S)-(-)-[pyridine-2-14C]nicotine hemisulfate (S-14CNHS), was synthesized by ChemSyn Science Laboratories (Lenexa, KS, USA) as a primary ethanol solution (630 µCi/ml) and was stored at -20 ± 5°C under an inert atmosphere protected from light. The stability of the test substance under these storage conditions and
of a representative dosing solution was determined by analysis of the chemical and radiochemical purity using reverse-phase HPLC with variable-wavelength and radiochemical detectors, and radio-optical purity was determined using a chiral HPLC method with variable-wavelength and radiochemical detectors.
- Analytical purity: Chemical purity was determined using reverse-phase high-performance liquid chromatography (HPLC) with a variable-wavelength detector. Purity was evaluated periodically during the course of testing. Radiochemical and radio-optical purities of the labeled S-14CNHS were 99% or greater throughout all the studies.
- Other: (S)-(-)-nicotine hemisulfate (S-NHS) was obtained from DeNovo (Lexington, KY, USA). All studies were performed at the Environmental Fate and Effects Laboratory of Roy F. Weston (West Chester, PA, USA). The S-NHS was stored at 4 ± 2°C in a septum-sealed, amber bottle with pressurized nitrogen in the headspace. Before initiation of the studies, the stability of the test substance under storage conditions for a representative dosing solution was determined by analysis of chemical and optical purity. Optical purity was determined using a chiral HPLC method with a variable-wavelength detector. Purity was evaluated periodically during the course of testing. Chemical and optical purities of the unlabeled S-NHS were 99% or greater throughout all the studies.
Radiolabelling:
yes
Oxygen conditions:
not specified
Soil classification:
not specified
Details on soil characteristics:
SOIL COLLECTION AND STORAGE
- Geographic location: Soil A was collected from an agricultural field in Spring City (PA, USA) and was considered to be unacclimated to nicotine. Soil B was collected from a site located in the R.J. Reynolds Tobacco Company Whitaker Park Manufacturing Complex in Winston-Salem and was considered to be acclimated to nicotine.
Soil No.:
#1
Duration:
28 d
Soil No.:
#2
Duration:
28 d
Soil No.:
#1
Initial conc.:
0.5 mg/kg soil d.w.
Based on:
test mat.
Soil No.:
#2
Initial conc.:
0.5 mg/kg soil d.w.
Based on:
test mat.
Parameter followed for biodegradation estimation:
test mat. analysis
Details on experimental conditions:
EXPERIMENTAL DESIGN
- Soil (g/replicate): 100 g dry weight/replicate
- Control conditions, if used (present differences from other treatments, i.e., sterile/non-sterile, experimental conditions): Three blanks were included for each soil used during the present study.
- No. of replication treatments: Triplicates
- Test apparatus (Type/material/volume): 500 ml Erlenmeyer flask was used as a test vessel for the study.
- Other details, if any: The flasks were aerated continuously during the course of the study with a CO2-free air source.

Test material application
- Volume of test solution used/treatment: 89 µl/treatment

SAMPLING DETAILS
- Method of collection of CO2 and volatile organic compounds: The 14CO2 released from the flasks was trapped in 1.5 N KOH. Samples were periodically collected and assayed for 14C activity by liquid scintillation counting.
Key result
Soil No.:
#1
% Degr.:
75.07
Parameter:
test mat. analysis
Sampling time:
28 d
Remarks on result:
other: % derg: 75.05 ± 5.1%, %degradation of test chemical was determined using the soil A
Key result
Soil No.:
#2
% Degr.:
76.54
Parameter:
test mat. analysis
Sampling time:
28 d
Remarks on result:
other: % derg: 76.54 ± 0.51%, %degradation of test chemical was determined using the soil B
Transformation products:
not specified
Evaporation of parent compound:
not specified
Volatile metabolites:
not specified
Residues:
not specified
Details on results:
For test soil A (no known previous exposure to nicotine), the final percentage T14CO2 for the three flasks spiked with S-14CNHS averaged 75.05% ±5.1% (mean±standard deviation), whereas the triplicate flasks containing the reference substance (stearic acid) averaged 79.32%±1.6%.
 
For test soil B (previous exposure to nicotine), the final percentage T14CO2 averaged 76.54%±0.51% in the S-14CNHS flasks and 79.27%±2.7% in the reference flasks.

Table: Biodegradation of (S)-(-)-[pyridine-2-14C]nicotine hemisulfate (S-14CNHS) in soil as indicated by the percentage of theoretical 14CO2 (T14CO2)a.

 

Test substance

Conc. (mg/kg)

Soilb

Final T14CO2 (%)

Aqueous supernatant (% soluble 14C)c

Soil (% bound 14C)d

Mass balance (%)

S-14NHS

0.5

A

77.72

69.15

78.29

1.04

NA

NA

13.70

NA

NA

92.46

NC

NC

[14C]stearic acid

 

0.5

A

77.87

80.99

79.09

NA

1.23

NA

NA

21.92

NA

NC

104.14

NC

S-14NHS

0.5

B

76.27

77.13

76.21

NA

NA

1.10

NA

NA

21.01

NC

NC

98.32

14C]stearic acid

0.5

B

81.54

79.92

76.35

NA

1.00

NA

NA

31.60

NA

NC

112.52

NC

 

Conclusions:
The percentage degradation of test chemical 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid in soil A and B was determined to be 75.05 ± 5.1% and 76.54 ± 0.51% in 28 days, respectively.
Executive summary:

Biodegradation study in soil was conducted for 28 days for determining the half-life value of the chemical 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid (CAS no. 65-35-0). The study was performed using theModified OECD Guideline 304 A (Inherent Biodegradability in Soil) in sludge-composited agricultural soil and in collected soil. Radiolabeling of test chemical was done at the carbon position.The test substance, radiolabeled (S)-(-)-[pyridine-2-14C]nicotine hemisulfate (S-14CNHS), was synthesized by ChemSyn Science Laboratories (Lenexa, KS, USA) as a primary ethanol solution (630µCi/ml) and was stored at-20±5°C under an inert atmosphere protected from light. Thestability of the test substance under these storage conditions and of a representative dosing solution was determined by analysis of the chemical and radiochemical purity using reverse-phase HPLC with variable-wavelength and radiochemical detectors, and radio-optical purity was determined using a chiral HPLC method with variable-wavelength and radiochemical detectors.Chemical purity was determined using reverse-phase high-performance liquid chromatography (HPLC) with a variable-wavelength detector.Purity was evaluated periodically during the course of testing. Radiochemical and radio-optical purities of the labeled S-14CNHS were 99% or greater throughout all the studies.Soil A was collected from an agricultural field in Spring City (PA, USA) and was considered to be unacclimated to nicotine. Soil B was collected from a site located in the R.J. Reynolds Tobacco Company Whitaker Park Manufacturing Complex in Winston-Salem and was considered to be acclimated to nicotine. Test chemical concentration used for the study was0.5 mg/kg.The soil A test consisted of eight glass, 500-ml Erlenmeyer flasks, each containing 100 g dry weight equivalent of a soil test system. The test (89µl) and reference ([14C]stearic acid, 120µl) substances were each dosed in triplicate flasks for each soil at a concentration of 0.5 mg/kg, and the water content was adjusted to approximately 35% using deionized water. The soil B test consisted of eight glass, 500-ml Erlenmeyer flasks, each containing 100 g dry weight equivalent of a soil test system. The test (89µl) and reference ([14C]stearic acid, 120µl) substances were each dosed in triplicate flasks for each soil (A and B) at a concentration of 0.5mg/kg, and the water content was adjusted to approximately 35% using deionized water.In addition, three blanks were included for each soil used during the present study. The flasks were aerated continuously during the course of the study with a CO2-free air source.The 14CO2 released from the flasks was trapped in 1.5 N KOH. Samples were periodically collected and assayed for 14C activity by liquid scintillation counting.After 28 d, the contents of the flasks were acidified with 1 N HCl and allowed to incubate for 3 d. A final set of samples was collected and analyzed on day 31, and a mass-balance determination was performed on a single flask from each treatment.The percentage degradation of test chemical3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acidin soil A and B was determined to be 75.05 ± 5.1% and 76.54 ± 0.51% in 28 days, respectively. Thus, based on percentage degradation, it is concluded that the chemical 3 -[(2S)-1 -methylpyrrolidin-2 -yl]pyridine; sulfuric acidis not persistent in the soil environment and the exposure risk to soil dwelling animals is moderate to low.

Description of key information

Biodegradation study in soil was conducted for 28 days for determining the half-life value of the chemical 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid (CAS no. 65-35-0) (JOEL A. SECKAR et. al., 2008). The study was performed using theModified OECD Guideline 304 A (Inherent Biodegradability in Soil) in sludge-composited agricultural soil and in collected soil. Radiolabeling of test chemical was done at the carbon position.The test substance, radiolabeled (S)-(-)-[pyridine-2-14C]nicotine hemisulfate (S-14CNHS), was synthesized by ChemSyn Science Laboratories (Lenexa, KS, USA) as a primary ethanol solution (630µCi/ml) and was stored at-20±5°C under an inert atmosphere protected from light. Thestability of the test substance under these storage conditions and of a representative dosing solution was determined by analysis of the chemical and radiochemical purity using reverse-phase HPLC with variable-wavelength and radiochemical detectors, and radio-optical purity was determined using a chiral HPLC method with variable-wavelength and radiochemical detectors.Chemical purity was determined using reverse-phase high-performance liquid chromatography (HPLC) with a variable-wavelength detector.Purity was evaluated periodically during the course of testing. Radiochemical and radio-optical purities of the labeled S-14CNHS were 99% or greater throughout all the studies.Soil A was collected from an agricultural field in Spring City (PA, USA) and was considered to be unacclimated to nicotine. Soil B was collected from a site located in the R.J. Reynolds Tobacco Company Whitaker Park Manufacturing Complex in Winston-Salem and was considered to be acclimated to nicotine. Test chemical concentration used for the study was0.5 mg/kg.The soil A test consisted of eight glass, 500-ml Erlenmeyer flasks, each containing 100 g dry weight equivalent of a soil test system. The test (89µl) and reference ([14C]stearic acid, 120µl) substances were each dosed in triplicate flasks for each soil at a concentration of 0.5 mg/kg, and the water content was adjusted to approximately 35% using deionized water. The soil B test consisted of eight glass, 500-ml Erlenmeyer flasks, each containing 100 g dry weight equivalent of a soil test system. The test (89µl) and reference ([14C]stearic acid, 120µl) substances were each dosed in triplicate flasks for each soil (A and B) at a concentration of 0.5mg/kg, and the water content was adjusted to approximately 35% using deionized water.In addition, three blanks were included for each soil used during the present study. The flasks were aerated continuously during the course of the study with a CO2-free air source.The 14CO2 released from the flasks was trapped in 1.5 N KOH. Samples were periodically collected and assayed for 14C activity by liquid scintillation counting.After 28 d, the contents of the flasks were acidified with 1 N HCl and allowed to incubate for 3 d. A final set of samples was collected and analyzed on day 31, and a mass-balance determination was performed on a single flask from each treatment.The percentage degradation of test chemical3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acidin soil A and B was determined to be 75.05 ± 5.1% and 76.54 ± 0.51% in 28 days, respectively. Thus, based on percentage degradation, it is concluded that the chemical 3 -[(2S)-1 -methylpyrrolidin-2 -yl]pyridine; sulfuric acidis not persistent in the soil environment and the exposure risk to soil dwelling animals is moderate to low.

Key value for chemical safety assessment

Additional information

Biodegradation study in soil was conducted for 28 days for determining the half-life value of the chemical 3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid (CAS no. 65-35-0) (JOEL A. SECKAR et. al., 2008). The study was performed using theModified OECD Guideline 304 A (Inherent Biodegradability in Soil) in sludge-composited agricultural soil and in collected soil. Radiolabeling of test chemical was done at the carbon position.The test substance, radiolabeled (S)-(-)-[pyridine-2-14C]nicotine hemisulfate (S-14CNHS), was synthesized by ChemSyn Science Laboratories (Lenexa, KS, USA) as a primary ethanol solution (630µCi/ml) and was stored at-20±5°C under an inert atmosphere protected from light. Thestability of the test substance under these storage conditions and of a representative dosing solution was determined by analysis of the chemical and radiochemical purity using reverse-phase HPLC with variable-wavelength and radiochemical detectors, and radio-optical purity was determined using a chiral HPLC method with variable-wavelength and radiochemical detectors.Chemical purity was determined using reverse-phase high-performance liquid chromatography (HPLC) with a variable-wavelength detector.Purity was evaluated periodically during the course of testing. Radiochemical and radio-optical purities of the labeled S-14CNHS were 99% or greater throughout all the studies.Soil A was collected from an agricultural field in Spring City (PA, USA) and was considered to be unacclimated to nicotine. Soil B was collected from a site located in the R.J. Reynolds Tobacco Company Whitaker Park Manufacturing Complex in Winston-Salem and was considered to be acclimated to nicotine. Test chemical concentration used for the study was0.5 mg/kg.The soil A test consisted of eight glass, 500-ml Erlenmeyer flasks, each containing 100 g dry weight equivalent of a soil test system. The test (89µl) and reference ([14C]stearic acid, 120µl) substances were each dosed in triplicate flasks for each soil at a concentration of 0.5 mg/kg, and the water content was adjusted to approximately 35% using deionized water. The soil B test consisted of eight glass, 500-ml Erlenmeyer flasks, each containing 100 g dry weight equivalent of a soil test system. The test (89µl) and reference ([14C]stearic acid, 120µl) substances were each dosed in triplicate flasks for each soil (A and B) at a concentration of 0.5mg/kg, and the water content was adjusted to approximately 35% using deionized water.In addition, three blanks were included for each soil used during the present study. The flasks were aerated continuously during the course of the study with a CO2-free air source.The 14CO2 released from the flasks was trapped in 1.5 N KOH. Samples were periodically collected and assayed for 14C activity by liquid scintillation counting.After 28 d, the contents of the flasks were acidified with 1 N HCl and allowed to incubate for 3 d. A final set of samples was collected and analyzed on day 31, and a mass-balance determination was performed on a single flask from each treatment.The percentage degradation of test chemical3-[(2S)-1-methylpyrrolidin-2-yl]pyridine; sulfuric acid in soil A and B was determined to be 75.05 ± 5.1% and 76.54 ± 0.51% in 28 days, respectively. Thus, based on percentage degradation, it is concluded that the chemical 3 -[(2S)-1 -methylpyrrolidin-2 -yl]pyridine; sulfuric acidis not persistent in the soil environment and the exposure risk to soil dwelling animals is moderate to low.