Sample purification for the analysis of caffeine in tobacco by gas chromatography-mass spectrometry.

A commonly used additive to tobacco products is cocoa. A sensitive an selective method was developed to measure caffeine, a marker for cocoa, in tobacco by using gas chromatography-mass spectrometry (GC-MS). Tobacco components usually produce high background signals in GC-MS analysis. Therefore, a series of extraction steps were designed to effectively purify the tobacco extracts. The analytical recovery of caffeine was 100 when [trimethyl-13C3] caffeine was used as an isotope-dilution reference. A linear calibration curve was generated with caffeine concentration ranging from 0.01 to 20 micrograms/ml. The detection limit of caffeine was 0.02 microgram/ml in the final solution. This method was applied to several commercial tobacco products, of which the corresponding caffeine levels varied from below the detection limit to 125 micrograms/g.