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Cotinine is the primary metabolite of nicotine and the preferred biomarker for assessing cigarette smoke exposure. Several liquid chromatography–tandem mass spectrometry (LC–MS-MS) methods have been described for measuring cotinine in biological fluids. Sample preparation typically involves manual solvent evaporation and reconstitution steps. This study describes a novel LC–MS-MS method for the quantification of cotinine by using electrospray ionization with multiple reaction monitoring and cotinine- d  3  as internal standard, coupled with an automated solid-phase extraction (SPE) procedure. The assay was linear over the analytical range of 0.5–1,000 ng/mL. The limits of detection and quantification were 0.13 and 0.20 ng/mL, respectively. Intra-assay and inter-assay imprecision of cotinine in all samples was &lt;5 and &lt;10% (coefficient of variation), respectively. The analytical recovery of cotinine spiked into plasma was &gt;95–100%. Matrix effects in serum and plasma were &lt;10%.

Determination of Cotinine by LC–MS-MS with Automated Solid-Phase Extraction