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Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine by High Throughput Liquid Chromatography-Mass Spectrometry.
Chem Res Toxicol. 2016 Mar 21; 29(3):390-7.CR

Abstract

Nicotine is the most abundant alkaloid in tobacco accounting for 95% of the alkaloid content. There are also several minor tobacco alkaloids; among these are nornicotine, anatabine, and anabasine. We developed and applied a 96 well plate-based capillary LC-tandem mass spectrometry method for the analysis of nornicotine, anatabine, and anabasine in urine. The method was validated with regard to accuracy and precision. Anabasine was quantifiable to low levels with a limit of quantitation (LOQ) of 0.2 ng/mL even when nicotine, which is isobaric, is present at concentrations >2500-fold higher than anabasine. This attribute of the method is important since anatabine and anabasine in urine have been proposed as biomarkers of tobacco use for individuals using nicotine replacement therapies. In the present study, we analyzed the three minor tobacco alkaloids in urine from 827 smokers with a wide range of tobacco exposures. Nornicotine (LOQ 0.6 ng/mL) was detected in all samples, and anatabine (LOQ, 0.15 ng/mL) and anabasine were detected in 97.7% of the samples. The median urinary concentrations of nornicotine, anatabine, and anabasine were 98.9, 4.02, and 5.53 ng/mL. Total nicotine equivalents (TNE) were well correlated with anatabine (r(2) = 0.714) and anabasine (r(2) = 0.760). TNE was most highly correlated with nornicotine, which is also a metabolite of nicotine. Urine samples from a subset of subjects (n = 110) were analyzed for the presence of glucuronide conjugates by quantifying any increase in anatabine and anabasine concentrations after β-glucuronidase treatment. The median ratio of the glucuronidated to free anatabine was 0.74 (range, 0.1 to 10.9), and the median ratio of glucuronidated to free anabasine was 0.3 (range, 0.1 to 2.9). To our knowledge, this is the largest population of smokers for whom the urinary concentrations of these three tobacco alkaloids has been reported.

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Authors+Show Affiliations

von Weymarn LB
Department of Biochemistry Molecular Biology and Biophysics and Masonic Cancer Center, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Thomson NM
Department of Biochemistry Molecular Biology and Biophysics and Masonic Cancer Center, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Donny EC
Department of Psychology, University of Pittsburgh , Pittsburgh, Pennsylvania 15260, United States.
Hatsukami DK
Masonic Cancer Center and Department of Psychiatry, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Murphy SE
Department of Biochemistry Molecular Biology and Biophysics and Masonic Cancer Center, University of Minnesota , Minneapolis, Minnesota 55455, United States.

MeSH

AdolescentAlkaloidsAnabasineChromatography, High Pressure LiquidHumansMolecular StructureNicotinePyridinesSmokingTandem Mass SpectrometryTobacco

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

26825008

Citation

von Weymarn, Linda B., et al. "Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine By High Throughput Liquid Chromatography-Mass Spectrometry." Chemical Research in Toxicology, vol. 29, no. 3, 2016, pp. 390-7.
von Weymarn LB, Thomson NM, Donny EC, et al. Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine by High Throughput Liquid Chromatography-Mass Spectrometry. Chem Res Toxicol. 2016;29(3):390-7.
von Weymarn, L. B., Thomson, N. M., Donny, E. C., Hatsukami, D. K., & Murphy, S. E. (2016). Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine by High Throughput Liquid Chromatography-Mass Spectrometry. Chemical Research in Toxicology, 29(3), 390-7. https://doi.org/10.1021/acs.chemrestox.5b00521
von Weymarn LB, et al. Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine By High Throughput Liquid Chromatography-Mass Spectrometry. Chem Res Toxicol. 2016 Mar 21;29(3):390-7. PubMed PMID: 26825008.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitation of the Minor Tobacco Alkaloids Nornicotine, Anatabine, and Anabasine in Smokers' Urine by High Throughput Liquid Chromatography-Mass Spectrometry. AU - von Weymarn,Linda B, AU - Thomson,Nicole M, AU - Donny,Eric C, AU - Hatsukami,Dorothy K, AU - Murphy,Sharon E, Y1 - 2016/02/12/ PY - 2016/1/31/entrez PY - 2016/1/31/pubmed PY - 2017/1/28/medline SP - 390 EP - 7 JF - Chemical research in toxicology JO - Chem Res Toxicol VL - 29 IS - 3 N2 - Nicotine is the most abundant alkaloid in tobacco accounting for 95% of the alkaloid content. There are also several minor tobacco alkaloids; among these are nornicotine, anatabine, and anabasine. We developed and applied a 96 well plate-based capillary LC-tandem mass spectrometry method for the analysis of nornicotine, anatabine, and anabasine in urine. The method was validated with regard to accuracy and precision. Anabasine was quantifiable to low levels with a limit of quantitation (LOQ) of 0.2 ng/mL even when nicotine, which is isobaric, is present at concentrations >2500-fold higher than anabasine. This attribute of the method is important since anatabine and anabasine in urine have been proposed as biomarkers of tobacco use for individuals using nicotine replacement therapies. In the present study, we analyzed the three minor tobacco alkaloids in urine from 827 smokers with a wide range of tobacco exposures. Nornicotine (LOQ 0.6 ng/mL) was detected in all samples, and anatabine (LOQ, 0.15 ng/mL) and anabasine were detected in 97.7% of the samples. The median urinary concentrations of nornicotine, anatabine, and anabasine were 98.9, 4.02, and 5.53 ng/mL. Total nicotine equivalents (TNE) were well correlated with anatabine (r(2) = 0.714) and anabasine (r(2) = 0.760). TNE was most highly correlated with nornicotine, which is also a metabolite of nicotine. Urine samples from a subset of subjects (n = 110) were analyzed for the presence of glucuronide conjugates by quantifying any increase in anatabine and anabasine concentrations after β-glucuronidase treatment. The median ratio of the glucuronidated to free anatabine was 0.74 (range, 0.1 to 10.9), and the median ratio of glucuronidated to free anabasine was 0.3 (range, 0.1 to 2.9). To our knowledge, this is the largest population of smokers for whom the urinary concentrations of these three tobacco alkaloids has been reported. SN - 1520-5010 UR - https://www.unboundmedicine.com/medline/citation/26825008/Quantitation_of_the_Minor_Tobacco_Alkaloids_Nornicotine_Anatabine_and_Anabasine_in_Smokers'_Urine_by_High_Throughput_Liquid_Chromatography_Mass_Spectrometry_ DB - PRIME DP - Unbound Medicine ER -