Abstract: To simultaneously determine the Amadori compounds in different types of tobacco, the Maillard model reactions for 12 Amadori compounds without standard reagents on sale were conducted respectively. Based on high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS), the 12 Amadori compounds were qualitatively identified in the model reaction systems after being matched with accurate molecular weight and confirmed with the secondary fragmentation modes of targets. The quantitative and qualitative ion pairs for MRM mode were also selected. Based on our previously established external standard quantitative method for the 10 Amadori compounds with available standard reagents, the 12 Amadori compounds without standard reagents were approximately quantified by HPLC-MS/MS using the standard curves of other targets with available standard reagents and similar retention times and structures. The results showed that: 1) With more than 300 injections in successive six days, the inter-day precisions of 14 Amadori compounds were lower than 10%, and that of 5 Amadori compounds were in the range of 10%-15%. 2) The test results of different tobacco leaf samples indicated that the total weights of 22 Amadori compounds accounted for 2.47% in flue-cured tobacco leaves, 1.70% in oriental tobacco leaves, and only 0.14% in burley tobacco leaves. The concentrations of the 22 Amadori compounds differed greatly in the tobacco leaves of different types. This method could achieve rapid and simultaneous determination of 22 Amadori compounds in tobacco.