Abstract: In this article, the contents of solanesol in different parts of tobacco leaves, from different breeds and different planting areas were determined. The solanesol was extracted from tobacco leaves via saponification and ultrasonic assist extraction and determined by RP-HPLC and APCI-MS with linear range of 41.2～6180 ng(R2=0.9999). The precision of the retention time is 0.08%RSD (n=5) and the precision of the peak area determination is 0.20%RSD (n=5). The recovery of the method is 83.1%～103.9% with the detection limit of 3.72 ng at 211 nm for solanesol. The results indicate that the contents of solanesol in different parts of tobacco leaves are distinctly different. The content of solanesol in a single tobacco leave drops from the top part to the nether part. The content of solanesol in the peduncle is very low. In order to increase the production and cut the cost, the obacco leaves with higher solanesol should be selected for solanesol production.