Abstract: The main purpose of this paper was to explore the difference of aroma components among flue-cured tobacco leaves in Guizhou tobacco-growing areas and classify different regions accordingly. Seventeen aroma components indexes were analyzed by nhexane solvent extraction and liquid chromatography-gas chromatography-mass spectrometry (LC-GC-MS) method with flue-cured tobacco samples collected from 44 sampling points in ten production regions in Guizhou. The results showed the difference of aroma component contents among different varieties was small, while significant difference was observed among different regions and among different leaf positions. Moreover, fifteen aroma component indexes were significantly different among middle leaves from different regions in Guizhou. Principal component analysis indicated that megastigmatrienone, solanone and norsolandione et al. were the key indexes of aroma components in Guizhou tobacco-growing areas. The leaf collections were divided into three categories according to the different aroma components in tobacco samples. The first category included Dejiang, Xingren, Xixiu, Daozhen and Weining regions with lower content of aroma components. The second one included Guiding, Kaili, Kaiyang and Yuqing regions with higher content of geranyl acetone, megastigmatrienone, 3-hydroxy-β-damascone, neophytadiene and farnesylacetone. The third category was Panxian regions with higher content of solanone and norsolandione.