Determination of acrylamide in tobacco smoke and smokeless tobacco products by liquid chromatography tandem mass spectrometry
Acrylamide forms when carbohydrate-containing foods are fried, baked, or roasted at high temperatures and may cause cancer in laboratory animals at high doses. Although LC/MS methods have been developed for the analysis of acrylamide in food, there is little information on tobacco-related applications. In this project, a sensitive, selective and robust LC-MS/MS method was developed and validated for the determination of acrylamide in tobacco related matrices. Due to the complexity of the sample matrix and low expected levels of analyse, extensive sample clean-up steps were found to be necessary. Two grams of the ground sample were extracted with 20mL water and filtered. An aliquot of the filtrate was then washed with dichloromethane (DCM) and the aqueous portion subjected to two solid phase extractions (SPE); Bond Elut AccuCAT (mixed-mode strong cation and strong anion exchange resin) followed by Lichrolut-EN (polymeric resin). The final eluate was analyzed by LC-MS/MS with positive ESI under MRM mode. Quantification of acrylamide was accomplished using three MS/MS-ion transitions (72/55, 72/54, and 72/44) to enhance the method's selectivity. The limit of quantification (LOQ) using the signal-to-noise ratio (S/N =10) of processed tobacco samples was 50 ng/g. Method precision, based on the relative standard deviation (RSD) of 12 replicates for KR2R4F, was less than 15%. The method accuracy was evaluated using 3 levels of fortified sample performed in triplicate and resulted in recoveries ranging from 92 to 107 %. Since the acrylamide content of cigarette smoke is considerably higher, the method is also applicable to diluted cigarette smoke extracts without further clean up.