Solanesol is an important secondary metabolite in Nicotiana tabacum. Distribution of solanesol in Nicotiana tabacum was investigated by High Performance Liquid Chromatography (HPLC) method. The quantitative distribution of solanesol in various organs and tissues of N. Tabacum showed that solanesol content, obviously different in all organs, was 6.8, 18.3, 27.5, 45.8, and 68.0 times higher in leaves than that in the stalks, flowers, seeds, fruits and roots, respectively. The contents of solanesol in various parts of leaf, stalk and flower were determined. The content of solanesol in top leaf, middle leaf and bottom leaf gradually decreased (6.124, 5.813 and 5.687 mg·g-1, respectively) and the content of solanesol in various leaf-parts (leaf apex, leaf middle and leaf base) also gradually decreased. The content of solanesol in top stalk was 1.19 times and 1.92 times higher than that in the middle stalk and the bottom stalk, respectively. The content of solanesol in various tissues of stalk (epidermis, cortex and stele) dramatically decreased. The sepal contained higher concentration of solanesol (1.192 mg·g-1) compared to any other parts in flower. The study will provide the base data for the regulation and control of solanesol,moreover, it will provide the scientific evidences for the rational development and utilization of N. Tabacum resources.