Determination of the nicotine metabolites cotinine and trans-3'-hydroxycotinine in biologic fluids of smokers and non-smokers using liquid chromatography-tandem mass spectrometry: biomarkers for tobacco smoke exposure and for phenotyping cytochrome P450 2A6 activity

Abstract

The nicotine metabolite cotinine is widely used to assess the extent of tobacco use in smokers, and secondhand smoke exposure in non-smokers. The ratio of another nicotine metabolite, trans-3'-hydroxycotinine, to cotinine in biofluids is highly correlated with the rate of nicotine metabolism, which is catalyzed mainly by cytochrome P450 2A6 (CYP2A6). Consequently, this nicotine metabolite ratio is being used to phenotype individuals for CYP2A6 activity and to individualize pharmacotherapies for tobacco addiction. In this paper we describe a highly sensitive liquid chromatography-tandem mass spectrometry method for determination of the nicotine metabolites cotinine and trans-3'-hydroxycotinine in human plasma, urine, and saliva. Lower limits of quantitation range from 0.02 to 0.1ng/mL. The extraction procedure is straightforward and suitable for large-scale studies. The method has been applied to several thousand biofluid samples for pharmacogenetic studies and for studies of exposure to low levels of secondhand smoke. Concentrations of both metabolites in urine of non-smokers with different levels of secondhand smoke exposure are presented.

© 2011 Elsevier B.V. All rights reserved.

Figures

Figure 1

Metabolism of nicotine to cotinine and trans-3′-hydroxycotinine

Figure 2

Chemical structures of internal standards

Figure 3

SRM chromatograms of non-smokers’ plasma

Figure 4

SRM chromatograms of non-smokers’ saliva

Figure 5

SRM chromatograms of non-smokers’ urine

Figure 6

Correlation of cotinine concentrations in smokers determined by GC (27, 29) with concentrations determined by the method described in this paper. The analytical data were used for a published study (44)