We describe a method for the quantitative analysis of nicotine, cotinine, trans-3′-hydroxy cotinine, nornicotine, and anabasine in urine, serum, and plasma using liquid chromatography-tandem mass spectrometry. A mix of deuterium-labeled internal standards (IS) is added to a specimen aliquot. The aliquot is extracted using mixed-mode solid phase extraction and eluted into an autosampler vial for injection into an LC-MS-MS system. An Atlantis silica column is used for LC separation in hydrophilic interaction mode. Tandem mass spectrometry detection is performed in positive ion mode with electrospray ionization and two multiple reaction monitoring (MRM) transitions monitored for each analyte and IS.
