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TSRC, Tob. Sci. Res. Conf., 2014, 68, abstr. 79

Determination of benzo[a]pyrene in smokeless tobacco products using gas chromatography-mass spectrometry

MERUVA N.(1); HUANG C.B.(1); WAGNER K.A.(1); ADAMS B.(2)
(1) Altria Client Services, Richmond, Virginia, USA; (2) Eurofins Lancaster Laboratories, Richmond, Virginia, USA

Benzo[a]pyrene (B[a]P) is a Harmful and Potentially Harmful Constituent (HPHC) found in tobacco products. As mandated by the Family Smoking Prevention and Tobacco Control Act, tobacco manufacturers and importers are required to report quantities of HPHCs to the United States Food and Drug Administration (FDA). The objective of this work was to develop a highly sensitive, selective and validated method for the determination of B[a]P in the smokeless tobacco products (STPs). The method consisted of extraction of the tobacco sample using methanol followed by purification of the tobacco extract using solid phase extraction (SPE). The eluent from SPE was evaporated to dryness and the sample was reconstituted using an organic solvent prior to Gas Chromatography-Mass Spectrometry (GC/MS) analysis. The use of SPE cleanup and the concentration step followed by selected ion monitoring (SIM) using GC-MS provided a highly sensitive and selective method for determination of trace levels of B[a]P in STPs. CORESTA reference products (CRP-1, CRP-2, CRP-3 and CRP-4) and Kentucky reference cigarette filler (3R4F) were used for method validation. All requirements for method validation were met including linearity, accuracy, precision, limits of detection (LOD), limits of quantitation (LOQ), method robustness, and standard and sample extract stability. For example, the linearity was demonstrated with a coefficient of determination of R2>0.995, the mean recovery for B[a]P was within 93%-107%, and the LOQ was 0.5 ng/g. The advantages of GC-MS procedure over HPLC-UV for determination of B[a]P in STPs will be discussed.