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