Abstract: To develop and enhance characteristics of fresh aroma note of different flue-cured tobaccos (redried leaves of Yuxi C3F and Xuchang C3F), the aroma components in mainstream smoke of flue-cured tobacco of robust and fresh flavor types were separated by semi-preparative high performance liquid chromatography. Eleven fresh (green) aroma components were identified and their releases in mainstream smoke were determined by a GC-MS method with internal standards. The odor activity values (OAVs) of the 11 components were calculated, the contributions of the 11 components for the fresh aroma were compared, and the fresh aroma note of the two types of flue-cured tobacco were analyzed on the basis of the OAVs. The results showed that:1) The total levels of the 11 fresh aroma components in mainstream smoke of Yuxi C3F and Xuchang C3F were 5.16 and 4.47 μg/cig respectively, in which the contents of 3-acetylpyridine and geranyl acetone were higher. 2) The OAV of 2-ethylhexanol was less than 1, its contribution to the fresh aroma note was minor, while the OAVs of β-damascenone and damascene were higher, they played important roles in demonstrating the fresh aroma note of mainstream smoke. 3) Besides 2-ethylhexyl alcohol, the total OAV of the other 10 fresh aroma components in mainstream smoke of Yuxi C3F was 8.91×10⁶, five times as that of Xuchang C3F, which confirmed the stylistic characteristic of fresh aroma note was more prominent in Yuxi C3F, it was in accordance with the results of sensory evaluation.