A sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of opiates and amphetamines in oral fluid was developed and validated. A simple sample preparation of oral fluid involving centrifugation and injection of 10 μL of the supernatant substantially reduced total analysis time. Owing to the basic nature of the drugs and their metabolites, LCMS/ MS analysis was performed by reverse-phase chromatography and electrospray ionization (ESI) which is a rugged and efficient ionization technique for basic drugs. Stable isotope-labeled internal standards were used in the analysis and the calibration to compensate for any matrix effects. The method was fully validated with the linearity of the developed method established in the concentration range of 1 to 500 ng/mL and the limits of detection and quantitation were 1 ng/mL for amphetamine, methamphetamine, morphine, codeine, 6-acetylmorphine and 6-acetylcodeine. The precision and accuracy were determined by spiking oral fluid samples at six concentration levels. For all analytes, the relative standard deviations of intra- and inter-day precision were 0.9 to 6.4% and 0.9 to 12.9%, respectively. The method was also successfully applied to the analyses of 34 oral fluid specimens collected from patients participating in the substitution therapy program.
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