Quantifying Nicotine and Metabolites

Concentrations of nicotine, cotinine and trans-3′ hydroxycotinine in plasma and urine samples were measured by liquid chromatography-tandem mass spectrometry, as described in detail previously.(7 (link)) Urine samples were assayed before and after deconjugation with a glucuronidase enzyme, as described previously.(3 (link)) The concentration before deconjugation represents the free (unconjugated) concentration, while the concentration after deconjugation represents the total (sum of free and conjugated) metabolite. In addition to nicotine, cotinine and 3HC and their glucuronides, we also measured nicotine N-oxide, cotinine N-oxide, nornicotine and its glucuronide and norcotinine and its glucuronide. Abbreviations used for nicotine and metabolites are as follows: nicotine = Nic; nicotine glucuronide = Nic-G; cotinine = Cot; cotinine glucuronide = Cot-G; trans 3′ hydroxycotinine = 3HC; trans 3′ hydroxycotinine glucuronide = 3HC-G

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Benowitz N.L., Dains K.M., Dempsey D., Yu L, & Jacob P I.I.I. (2010). Estimation of Nicotine Dose after Low Level Exposure Using Plasma and Urine Nicotine Metabolites. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 19(5), 1160-1166.

Publication 2010

Cotinine Cotinine n oxide Enzyme Glucuronidase Glucuronide Liquid chromatography Nicotine Nicotine glucuronide Norcotinine Nornicotine Oxide Plasma Tandem mass spectrometry Trans 3 hydroxycotinine Urine

Corresponding Organization : University of California, San Francisco

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Variable analysis

independent variables

Concentrations of nicotine, cotinine and trans-3' hydroxycotinine

dependent variables

Concentrations of nicotine, cotinine and trans-3' hydroxycotinine in plasma and urine samples
Concentrations of nicotine N-oxide, cotinine N-oxide, nornicotine and its glucuronide, and norcotinine and its glucuronide

control variables

Urine samples were assayed before and after deconjugation with a glucuronidase enzyme

controls

Positive control: Urine samples were assayed after deconjugation with a glucuronidase enzyme
Negative control: Urine samples were assayed before deconjugation with a glucuronidase enzyme

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