The phytochemical study of n-butanol and ethyl acetate extracts of Calotropis procera (Asclepiadaceae), proceeded to the separation, purification and identification of two flavonoides: isorhamnetin-3-O-rutinoside (1) and isorhamnetin-3-O-rubinobioside (2), which were characterized by the use of combined spectral methods (UVvisible, NMR1H) in addition of acid hydrolysis. The total phenolic quantification, the antioxidant and antibacterial activities were determined.