A Solid Phase Extraction Method for Determination of Nicotine in Serum and Urine by Isotope Dilution Gas Chromatography/Mass Spectrometry with Selected Ion Monitoring

A rapid method for measuring nicotine concentration in serum and urine is described. Deuterated nicotine is used as an internal standard. Nicotine and deuterated nicotine are extracted using a copolymeric-bonded phase silica column. The extract is analysed by gas chromatography coupled with mass spectrometry (GC/MS) operating in selected ion monitoring mode. The method has a lower limit of detection of approximately 2 μg/L and is linear to at least 2000 μg/L. Within-run percentage coefficients of variation (% CV) are < 4 in both assays over a nicotine concentration range of 10–2000 μg/L. Between-run %CV in the serum assay are 5.4, 5.2, 4.8 and 5.9, respectively, at nicotine concentrations of 10, 15, 25, and 50 μg/L. Between-run %CV in the urine assay are 5.9, 4.5, 2.7 and 5.2, respectively, at nicotine concentrations of 100, 250, 500, and 2000 μg/L. The absolute recovery of nicotine is 61±+6% (mean±SD) over the range of 10–250 μg/L. The assay has been used to measure serum nicotine concentrations and 24-h urinary excretion of nicotine to monitor the extent of replacement in subjects receiving transdermal nicotine therapy for smoking cessation.