A new compound kaempferol 3-O-alpha-L-(2"-E-p-coumaroyl)-rhamnopyranoside, as well as the known flavonoids, kaempferol 3-O-beta-D-(6"-E-p-coumaroyl)-glucopyranoside, kaempferol 3-O-alpha-L-(2",3"-di-E-p-coumaroyl)-rhamnopyranoside, and caffeic acid were obtained from the methanolic extract of Platanus orientalis L. buds. All the compounds were isolated by column chromatography and identified using 1H-NMR, 2D-1H-NMR (COSY), 1H-13C-NMR, and CIDMS techniques. Cytotoxic and antimicrobial studies were carried out in vitro against human cell lines and against Gram-positive and Gram-negative organisms.