Simultaneous quantification of ten Amadori compounds in tobacco using liquid chromatography with tandem mass...

Amadori compounds are aroma precursors formed in the initial phase of the Maillard reaction. Based on their similar structures, simultaneous quantification of more than six Amadori compounds in tobacco has not been reported yet. In this study, a simple and rapid method was developed to simultaneously quantify ten Amadori compounds including the isomers of Fructose-isoleucine and Fructose-leucine in tobacco. The separation was performed on an Atlantis T₃ column (2.1 × 250 mm, 5 μm) by gradient elution using acetonitrile and water as the mobile phases. The quantification method was systematically evaluated and proven to be sensitive and accurate. The linearity was good, with correlation coefficients of 0.9977-0.9999. The limits of detection and quantitation were 1.354-2.532 and 4.516-8.444 ng/mL, respectively. The recoveries were 84.0-119.6%, and the relative standard deviations were 1.33-5.40%. The method was used to analyze the changes in the amounts of ten Amadori compounds in tobacco before and after tobacco primary processing. The analysis shows that the Maillard reaction occurs during the short processing period.