Investigation on Ballota andreuzziana (Family Labiatae) for flavonoids led to isolation of two aglycones from ethyl acetate fraction which were identified as 7-methoxy luteolin and 6,7-dimethoxy scutellarein, in addition to three glycosides known as luteolin-7-O-glucoside, 6, 4'-dimethoxy scutellarein-7-O-glucoside and quercetin-7-O-rhamno-glucoside from butanol fraction. Antimicrobial studies revealed that, butanol extract exhibited the marked activity against M0. Phlei, S. aureus and C. albicans (I.Z. =16.3, 11.3, and 10.7mm, Conc.=150mg/ml) respectively. While, chloroform extract also showed the better activity against B. subtilis (I.Z. =11.0mm, Conc.=150mg/ml). Furthermore aqueous extracts exhibited no effects at all against all tested organisms at all concentrations also, all tested Gram negative bacteria and the fungus A. niger are resistance to all concentrations of tested extracts.