Determination of nicotine and four metabolites in the serum of smokers by high-performance liquid chromatography with ultraviolet detection.

P Zuccaro, I Altieri, M Rosa, A R Passa, S Pichini, G Ricciarello, R Pacifici

Index: J. Chromatogr. A. 621(2) , 257-61, (1993)

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Abstract

A simple and reliable reversed-phase high-performance liquid chromatographic method with ultraviolet detection is described for the quantitation of nicotine and its metabolites cotinine, trans-3'-hydroxycotinine, norcotinine and cotinine N-oxide in human serum. The analytes and the internal standard, N-ethylnorcotinine, were extracted by solid-phase extraction before chromatography. Two different columns and mobile phases with gradient systems were used. The detection limit of the assay was 10 ng/ml for nicotine, 3 ng/ml for cotinine N-oxide and 5 ng/ml for cotinine, trans-3'-hydroxycotinine and norcotinine. The concentrations of nicotine and its metabolites in the serum of 12 cigarette smokers are reported.