A novel dehydroxy gossypol was isolated from the methanol extract of bark of Thespesia populnea (Linn) by using the mass triggered preparative HPLC. The crude methanol extract of Thespesia populnea (Linn) was pre-purified by filtration through celite and column chromatography by using neutral alumina as a stationary phase and Ethyl acetate: Hexane as a mobile phase. Then the mixture was taken to the HPLC method development by buffer (Acidic and Neutral): Acetonitrile as a mobile phases and Xbridge (C18, 4.6 X 150 mm, 3.5 μm) and Sunfire (C18, 4.6 X 150 mm, 3.5 μm) as stationary phases. Finally the isolation was performed by using automated preparative HPLCMS with Xbridge column (C18, 4.6 X 150 mm, 5 μm). Then the isolated was major compound was characterized by 1H NMR, 13C NMR and LCMS.
