A method using gas chromatography (GC)-mass spectrometry ( MS) for the simultaneous determination of the smoke uptake parameters thiocyanate, nicotine and cotinine in human tissues is reported. Nicotine, cotinine and thiocyanate, in combination with a phase-transfer catalyst, were extracted from urine, saliva and hair into dichloromethane (DCM). Thiocyanate was alkylated in the DCM-layer to form a pentafluorobenzyl derivative. The biochemical markers in DCM were directly injected into the GC system and separated on a DB-1MS column using a 9.4 min temperature program. The method was validated in urine and saliva between the limits of quantitation (1.0-15 mug ml(-1) thiocyanate, 0.010 - 3.0 mug ml(-1) nicotine and cotinine in urine, 0.010 - 1.0 mug ml(-1) nicotine and cotinine in saliva). The calibration curves were found to be linear (r > 0.996), the within- and between-day accuracy's were 83 - 120%, the repeatability coefficients of variation were 3 - 20% and the limits of detection were 0.060 ng ml(-1) thiocyanate and 0.60 ng ml(-1) nicotine and cotinine. The results of the analysis of the biomarkers in the urine of 44 volunteers were used to develop a predictive model for smoking status, using discriminant analysis. The classification model correctly classified 93.2% of cross-validated grouped cases. Saliva samples were used to confirm the results of the classification method.
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