Amadori compounds are important aroma precursors. Therefore, rapid and corrective determination of Amadori compounds is meaningful for monitoring the quality of tobacco leaves. By the end of 2016, the publicated methods have analyzed no more than 6 Amadori compounds simultaneously in tobacco. And in 2017, our research group reported a HPLC- MS/MS method to simultaneously quantify 10 glucose derived Amadori compounds in tobacco. This paper is based on our previous study. With respect to another 12 Amadori compounds without commericial standards, model reactions were conducted. And by using HPLC-Q-TOF-MS with high mass resolution, the 12 Amadori compounds were identified in the model reaction systems based on being matched with accurate mass and confirmed with the fragmentation way. The quantitative and the qualitative ion pairs in MRM mode were also selected. Then using HPLC-MS/MS, we quantifyed the 10 Amadori compounds with standards using external standard quantitative method, and semi-quantified the 12 Amadori compounds without standards using the standard curves of others with similar retention time and abundance in tobacco. Through large sample test, except 4 compounds which were unstable or of abundance close to LOQ, the intra-day precisions of 18 Amadori compounds were lower than 10%. The inter-day precisions of 14 Amadori compounds were lower than 10%, and that of 4 compounds could be controlled between 10% and 15%. The developed method was used in testing tobacco leaves of different types. The results showed that the total weights of 22 Amadori compounds accounted for 2.4% in flue-cured tobacco, 1.7% in oriental tobacco, 0.14% in burley tobacco. And the content distributions of the 22 Amadori compounds were also different among different type of tobacco leaves.