Improved Methodology for the Quantitative Assessment of Tobacco-Specific N-Nitrosamines in Tobacco by Supercritical Fluid Extraction

This paper describes an analytical method for the assessment of tobacco-specific N-nitrosamines (TSNA) in tobacco which involves supercritical fluid extraction and quantification by gas chromatography with thermal energy analyzer detection. Extraction of tobacco with supercritical carbon dioxide, to which 10% methanol is added, is followed by a cleanup on silica cartridges which separates the TSNA from the bulk of other tobacco components present in the extract. Gas chromatographic separation is performed on a DB-5 fused silica capillary column, and quantitative results are ascertained by using 4-(ethylnitrosamino)-1-(3-pyridyl)-1-butanone as an internal standard. This method is quantitative and well reproducible and meets the qualifications to be chosen as a standardized analytical method.