Quantitation of Urine Nicotine, Cotinine, and 3-OH-Cotinine by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Nicotine is a naturally occurring and highly addictive chemical used in e-cigarettes, cigarettes, chewing tobacco, and other tobacco products as well as in nicotine replacement therapies. The negative health consequences of using nicotine-containing products are well known. In fact, smoking remains the leading cause of preventable disease, disability, and death in the United States. Measurement of nicotine and its metabolites, cotinine and 3-OH-cotinine, offers an objective method to evaluate nicotine exposure and the associated health risks. In this chapter, we describe a quick and reliable isotope dilution LC-MS/MS method for the quantitation of these three compounds in 60 μL of human urine following a simple sample preparation procedure. Electrospray Ionization (ESI) in positive mode is used to introduce the analytes into the mass spectrometer and quantitation is achieved using Multiple Reaction Monitoring (MRM). The analytical measurable ranges for nicotine and cotinine are 10–2500 ng/mL and 20–5000 ng/mL for 3-OH-cotinine.