Mass Spectrometric Quantitation of Nicotine, Cotinine, and 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanol in Human Toenails

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Abstract
Numerous studies have quantified total cotinine (the sum of cotinine and cotinine-N-glucuronide) and total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol [NNAL; the sum of NNAL and its O- and N-glucuronides (NNAL-Glucs)] in the urine and blood of smokers, smokeless tobacco users, and nonsmokers exposed to environmental tobacco smoke. Analysis of hair and nails has several advantages over blood and urine testing, such as accumulation of xenobiotics during long-term exposure, ease of collection, and indefinite stability of samples. We developed sensitive methods for quantitation of nicotine, cotinine, and NNAL in human toenails. Nicotine and cotinine were analyzed by gas chromatography-mass spectrometry-selected ion monitoring. NNAL was assayed using liquid chromatography-electrospray ionization-tandem mass spectrometry-selected reaction monitoring. The detection limits of the methods were 0.01 ng/mg toenail for nicotine, 0.012 ng/mg toenail for cotinine, and 0.02 pg/mg toenail for NNAL. In 35 smokers, the mean nicotine level was 5.9 ± 5.6 ng/mg toenail, mean cotinine was 1.6 ± 1.3 ng/mg toenail, and mean NNAL was 0.41 ± 0.67 pg/mg toenail. Samples collected from six nonsmokers were negative for NNAL. In smokers, NNAL correlated with cotinine (r = 0.77; P < 0.0001). The results of this study for the first time show the presence of cotinine and NNAL in human toenails. These sensitive and quantitative methods should be useful in epidemiologic studies of the role of chronic tobacco smoke exposure, including environmental tobacco smoke exposure, in human cancer. (Cancer Epidemiol Biomarkers Prev 2006;15(12):2378–83)

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