To investigate the pharmacodynamic substance basis of Blumea balsamifera (L.) DC., fifteen compounds were isolated from the EtOAc fraction by silica gel, sephadex LH-20 and ODS-A column chromatography. The structures were determined based on the analysis of the spectroscopic data as 3,3°,5-trihydroxy-4°,7-dimethoxy flavanone (1), 4°,5-dihydroxy-3,3°,7-trimethoxy flavone (2), blumeatin (3), 3,5,3°,4°-tethydroxy-7-methoxy flavone (4), diosmetin (5), 3°,4°,5-trihydroxyl-3,7-dimethoxy flavone (6), isosamicin (7), chrysosplenol C (8), hyperoside (9), isoquercitrin (10), 3°,5,7-trihydroxy-4°-methoxy flavanone (11), sakuranetin (12), pilloin (13), 5,7,3°,4°-tetrahydroxy-3-methoxyflavone (14) and 5-hydroxy-3,7,3°,4°-tetramethoxy flavone (15). Compounds 7, 11, 12 and 13 are isolated from the plant for the first time. Antibacterial test indicated that 1, 3, 6, 8 and 12 displayed different inhibition against three bacteria, and 3 significantly inhibited Staphylococcus aureus subsp. aureus (DSM 799) with a MIC value of 32 μg/mL.