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Simultaneous Determination of Nicotine and Its Nine Metabolites in Human Urine by LC–MS–MS

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Abstract

A method based on liquid chromatography tandem mass spectrometry was developed for the direct determination of nicotine, cotinine, trans-3′-hydroxycotinine, their corresponding glucuronide conjugates as well as nornicotine, norcotinine, cotinine-N-oxide and nicotine-N-oxide in the urine of smokers. The assay only involves centrifugation and filtration of diluted urine. The analysis was performed on a C18 reversed-phase column using a gradient of 10 mM ammonium acetate, pH 6.8, and methanol as mobile phase at a flow rate of 1 mL min−1. Nicotine-methyl-d3, Cotinine-methyl-d3 and trans-3′-hydroxycotinine-methyl-d3 were used as internal standards. Precisions (RSD) for all the analytes at three levels were between 2.1 and 17.0%. Recoveries for nicotine and nine nicotine metabolites ranged from 78.4 to 115.6%. The described method was suitable for determining the nicotine dose in large-scale human biomonitoring studies.

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Acknowledgment

Financial support from the National Nature Science Foundation of China (No. 20705040) and the Scientific Foundation of State Tobacco Monopoly Administration of China is gratefully acknowledged.

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Correspondence to Huimin Liu.

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Fan, Z., Xie, F., Xia, Q. et al. Simultaneous Determination of Nicotine and Its Nine Metabolites in Human Urine by LC–MS–MS. Chroma 68, 623–627 (2008). https://doi.org/10.1365/s10337-008-0729-9

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  • DOI: https://doi.org/10.1365/s10337-008-0729-9

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