Abstract
Nicotine is a naturally occurring and highly addictive chemical used in e-cigarettes, cigarettes, chewing tobacco, and other tobacco products as well as in nicotine replacement therapies. The negative health consequences of using nicotine-containing products are well known. In fact, smoking remains the leading cause of preventable disease, disability, and death in the United States. Measurement of nicotine and its metabolites, cotinine and 3-OH-cotinine, offers an objective method to evaluate nicotine exposure and the associated health risks. In this chapter, we describe a quick and reliable isotope dilution LC-MS/MS method for the quantitation of these three compounds in 60 μL of human urine following a simple sample preparation procedure. Electrospray Ionization (ESI) in positive mode is used to introduce the analytes into the mass spectrometer and quantitation is achieved using Multiple Reaction Monitoring (MRM). The analytical measurable ranges for nicotine and cotinine are 10–2500 ng/mL and 20–5000 ng/mL for 3-OH-cotinine.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Mokdad AH, Marks JS, Stroup DF et al (2004) Actual causes of death in the United States, 2000. JAMA 291:1238–1245
Kondo T, Nakano Y, Adachi S et al (2019) Effects of tobacco smoking on cardiovascular disease. Circ J 83:1980–1985
Benowitz NL (2009) Pharmacology of nicotine: addiction, smoking-induced disease, and therapeutics. Annu Rev Pharmacol Toxicol 49:57–71
Benowitz NL, Jacob P 3rd, Fong I et al (1994) Nicotine metabolic profile in man: comparison of cigarette smoking and transdermal nicotine. J Pharmacol Exp Ther 268:296–303
Mallock N, Rabenstein A, Peter L et al (2021) Rapid sensitive, and reliable quantitation of nicotine and its main metabolites cotinine and trans-3′-hydroxycotinine by LC-MS/MS: method development and validation for human plasma. J Chromatogr B Analyt Technol Biomed Life Sci 1179:122736
Nguyen TD, Riordan-Short S, Dang TT et al (2020) Quantitation of select terpenes/terpenoids and nicotine using gas chromatography-mass spectrometry with high-temperature headspace sampling. ACS Omega 5:5565–5573
Stepanov I, Feuer R, Jensen J et al (2006) Mass spectrometric quantitation of nicotine, cotinine, and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol in human toenails. Cancer Epidemiol Biomark Prev 15:2378–2383
Shu I, Wang P (2013) Simultaneous serum nicotine, cotinine, and trans-3′-hydroxycotinine quantitation with minimal sample volume for tobacco exposure status of solid organ transplant patients. J Chromatogr B Analyt Technol Biomed Life Sci 928:139–145
Yue B, Kushnir MM, Urry FM et al (2010) Quantitation of nicotine, its metabolites, and other related alkaloids in urine, serum, and plasma using LC-MS-MS. Method Mol Biol 603:389–398
Clarke MB (2002) Quantitation of nicotine in tobacco products by capillary electrophoresis. J AOAC Int 85:1–7
Moyer TP, Charlson JR, Enger RJ et al (2002) Simultaneous analysis of nicotine, nicotine metabolites, and tobacco alkaloids in serum or urine by tandem mass spectrometry, with clinically relevant metabolic profiles. Clin Chem 48:1460–1471
Miller EI, Norris HRK, Rollins DE et al (2010) A novel validated procedure for the determination of nicotine, eight nicotine metabolites and two minor tobacco alkaloids in human plasma or urine by solid-phase extraction coupled with liquid chromatography-electrospray ionization-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 878:725–737
Hoofnagle AN, Laha TJ, Rainey PM et al (2006) Specific detection of anabasine, nicotine, and nicotine metabolites in urine by liquid chromatography-tandem mass spectrometry. Am J Clin Pathol 126:880–887
McGuffey JE, Wei B, Bernert JT et al (2014) Validation of a LC-MS/MS method for quantifying urinary nicotine, six nicotine metabolites and the minor tobacco alkaloids-anatabine and anabasine- in smokers’ urine. PLoS One 9:e101816
Oh J, Park MS, Chun MR et al (2022) A simple and high-throughput LC-MS-MS method for simultaneous measurement of nicotine, cotinine, 3-OH cotine, nornicotine and anabasine in urine and its application in the general Korean population. J Anal Toxicol 46:25–36
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Li, W., Zhang, X.S., Noguez, J. (2024). Quantitation of Urine Nicotine, Cotinine, and 3-OH-Cotinine by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). In: Garg, U. (eds) Clinical Applications of Mass Spectrometry in Drug Analysis. Methods in Molecular Biology, vol 2737. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3541-4_31
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3541-4_31
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3540-7
Online ISBN: 978-1-0716-3541-4
eBook Packages: Springer Protocols