Abstract
We describe a method for the quantitative analysis of nicotine, cotinine, trans-3′-hydroxy cotinine, nornicotine, and anabasine in urine, serum, and plasma using liquid chromatography-tandem mass spectrometry. A mix of deuterium-labeled internal standards (IS) is added to a specimen aliquot. The aliquot is extracted using mixed-mode solid phase extraction and eluted into an autosampler vial for injection into an LC-MS-MS system. An Atlantis silica column is used for LC separation in hydrophilic interaction mode. Tandem mass spectrometry detection is performed in positive ion mode with electrospray ionization and two multiple reaction monitoring (MRM) transitions monitored for each analyte and IS.
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References
(1988) The health consequences of smoking – nicotine addiction: a report of the Surgeon General. USA Department of Health and Human Services.
(1999) Achievements in public health, 1900–1999: Tobacco Use – United States, 1900–1999. MMWR CDC Surveill.Summ. 48, 986–993.
(2004) The health consequences of smoking: a report of the Surgeon General. USA Department of Health and Human Services.
Jacob, P., Yu, L., Shulgin, A.T., and Benowitz, N.L. (1999) Minor tobacco alkaloids as biomarkers for tobacco use: comparison of users of cigarettes, smokeless tobacco, cigars, and pipes. Am. J. Public Health 89, 731–736.
Jacob, P., Hatsukami, D., Severson, H., Hall, S., Yu, L., and Benowitz, N.L. (2002) Anabasine and anatabine as biomarkers for tobacco use during nicotine replacement therapy. Cancer Epidemiol. Biomarkers Prev. 11, 1668–1673.
Xu, A.S., Peng, L.L., Havel, J.A., Petersen, M.E., Fiene, J.A., and Hulse, J.D. (1996) Determination of nicotine and cotinine in human plasma by liquid chromatography-tandem mass spectrometry with atmospheric-pressure chemical ionization interface. J. Chromatogr. B Biomed. Appl. 682, 249–257.
Bernert, J.T. Jr., Turner, W.E., Pirkle, J.L., Sosnoff, C.S., Akins, J.R., Waldrep, M.K., Ann, Q., Covey, T.R., Whitfield, W.E., Gunter, E.W. et al. (1997) Development and validation of sensitive method for determination of serum cotinine in smokers and nonsmokers by liquid chromatography/atmospheric pressure ionization tandem mass spectrometry. Clin. Chem. 43, 2281–2291.
Moyer, T.P., Charlson, J.R., Enger, R.J., Dale, L.C., Ebbert, J.O., Schroeder, D.R., and Hurt, R.D. (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.
Stolker, A.L., Niesing, W., Hogendoorn, E.A., Bisoen Rambali, A., and Vleeming, W. (2003) Determination of nicotine and cotinine in rat plasma by liquid chromatography-tandem mass spectrometry. J. Chromatogr. A 1020, 35–43.
Kellogg, M.D., Behaderovic, J., Bhalala, O., and Rifai, N. (2004) Rapid and simple tandem mass spectrometry method for determination of serum cotinine concentration. Clin. Chem. 50, 2157–2159.
Xu, X., Iba, M.M., and Weisel, C.P. (2004) Simultaneous and sensitive measurement of anabasine, nicotine, and nicotine metabolites in human urine by liquid chromatography-tandem mass spectrometry. Clin. Chem. 50, 2323–2330.
Kim, I., and Huestis, M.A. (2006) A validated method for the determination of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in human plasma using solid-phase extraction and liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry. J. Mass Spectrom. 41, 815–821.
Tuomi, T., Johnsson, T., and Reijula, K. (1999) Analysis of nicotine, 3-hydroxycotinine, cotinine, and caffeine in urine of passive smokers by HPLC-tandem mass spectrometry. Clin. Chem. 45, 2164–2172.
Meger, M., Meger-Kossien, I., Schuler-Metz, A., Janket, D., and Scherer, G. (2002) Simultaneous determination of nicotine and eight nicotine metabolites in urine of smokers using liquid chromatography-tandem mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 778, 251–261.
Heavner, D.L., Richardson, J.D., Morgan, W.T., and Ogden, M.W. (2005) Validation and application of a method for the determination of nicotine and five major metabolites in smokers' urine by solid-phase extraction and liquid chromatography-tandem mass spectrometry. Biomed. Chromatogr. 19, 312–328.
Hoofnagle, A.N., Laha, T.J., Rainey, P.M., and Sadrzadeh, S.M. (2006) Specific detection of anabasine, nicotine, and nicotine metabolites in urine by liquid chromatography-tandem mass spectrometry. Am. J. Clin. Pathol. 126, 880–887.
Bentley, M.C., Abrar, M., Kelk, M., Cook, J., and Phillips, K. (1999) Validation of an assay for the determination of cotinine and 3-hydroxycotinine in human saliva using automated solid-phase extraction and liquid chromatography with tandem mass spectrometric detection. J. Chromatogr. B Biomed. Sci. Appl. 723, 185–194.
Marchei, E., Joya, X., Garcia-Algar, O., Vall, O., Pacifici, R., and Pichini, S. (2008) Ultrasensitive detection of nicotine and cotinine in teeth by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom. 22, 2609–2612.
Gray, T.R., Shakleya, D.M., and Huestis, M.A. (2008) Quantification of nicotine, cotinine, trans-3'-hydroxycotinine, nornicotine and norcotinine in human meconium by liquid chromatography/tandem mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 863, 107–114.
Kushnir, M.M., Rockwood, A.L., Nelson, G.J., Yue, B., and Urry, F.M. (2005) Assessing analytical specificity in quantitative analysis using tandem mass spectrometry. Clin. Biochem. 38, 319–327.
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Yue, B., Kushnir, M.M., Urry, F.M., Rockwood, A.L. (2010). Quantitation of Nicotine, Its Metabolites, and Other Related Alkaloids in Urine, Serum, and Plasma Using LC-MS-MS. In: Garg, U., Hammett-Stabler, C. (eds) Clinical Applications of Mass Spectrometry. Methods in Molecular Biology, vol 603. Humana Press. https://doi.org/10.1007/978-1-60761-459-3_38
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DOI: https://doi.org/10.1007/978-1-60761-459-3_38
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