Analysis of Certain Nitrogenous Compounds in Tobacco. Part 1: Adenosine, 2,5- and 2,6-Deoxyfructosazines, Mannosamine and Glucosamine

Nitrogenous compounds such as amino acids and proteins are frequently analyzed in tobacco since they are considered precursors of toxicants in cigarette smoke. However, much less attention is given to other nitrogenous compounds such as amino sugars and deoxyfructosazines, although their concentration in tobacco can be equal to or even higher than that of most free amino acids. These nitrogenous compounds may contribute to the formation of toxicants in smoke, or may contribute to the sensory properties of cigarette smoke, reasons for which their analysis is important. This study describes a procedure for the analysis of adenosine, 2,5- and 2,6-deoxyfructosazines (DFs), mannosamine and glucosamine in tobacco. The analysis uses a liquid chromatographytandem mass spectrometry (LC/MS/MS) technique. Sample preparation for analysis consists of the extraction of the tobacco with a solution of 90% water and 10% methanol, followed by filtration. The separation of the analytes was done on a hydrophilic interaction liquid chromatography HILIC column using an isocratic procedure with a solvent consisting of 78% CH3CN, 22% H2O, that also contained 0.1 % HCOOH and 0.143 g/L CH3COONH4. The measurements were done using electrospray positive ionization mass spectrometric detection. The analytical procedure was validated and was proven very reliable. A number of tobaccos were analyzed, including several fluecured and Burley USA tobaccos, off-shore tobaccos, tw