The flavonoids rhamnocitrin (5,4’-dihydroxy-7-methoxyflavonol), sakuranetin (5,4’-dihydroxy-7-methoxyflavanone) and 3,8-dimethoxy-gossypetin (5,7,3’,4’-tetrahydroxy-3,8-dimethoxyflavone) were isolated and purified from Chromolaena subscandens’ leaves using column chromatography and thin layer chromatography methods. These substances were identified on the basis of their physical properties (melting point and Rf value), qualitative chemical reactions and spectroscopic techniques UV and NMR (experiments 1H, 13C, 1H-1H COSY, HMQC and HMBC), according to their complexity.