The antimicrobial activity of methanol extract of N erium indicum, Tagetus erecta, Chrysanthemum leucanthemum, Rosa centrifolia, Jasmi num angustifolium, Torenia fournieri against Gram negative and Gram positive bacteria we re studied invitro. The objective of this research was to confirm the antibacterial activity and perform HPTLC analysis of methanolic extracts of various flower extracts. The methanolic extracts of Nerium indicum, Tagetus erecta, Rosa centrifolia, Torenia fournieri exhibited growt h inhibition on selected bacterial strains viz., Bacillus sp., Escherichia coli., Klebsiella sp., Ye rsinia sp., Enterococcus sp. Based on the results, the methanol extract of Rosa centrifolia w as considered to be the most effective and also indicated that all the flower extracts exhibited in hibitory action against the growth of Lactobacillus sp., whereas Chrysanthemum leucanthem um, Jasminum angustifolium, did not show any inhibition on the test bacterial species. Further the extracts were separated on the TLC plates on selected mobile phase and analysed in HPT LC using CAMAG software.