Abstract: In order to investigate the best extraction methods of leaf trichome exudates in flue-cured tobacco, three flued-cured tobacco varieties including “K326”, “Yunyan85” and “honghuadajinyuan”, were used in this study. Effects of different extraction degrees and different lengths of extraction time on leaf weight changes were compared. The results showed that the significant change of weight was found before the third extraction in three flue-cured tobacco varieties, while it’s no significant different in leaf weight change following the increases in extraction degrees. The effect of different extraction degrees on leaf weight change seemed little. This indicates that the three degrees extraction method of leaf trichome exudates in flue-cured tobacco is more effective, which will be useful in study on leaf trichome exudates in flue-cured tobacco for the future.