Propolis is a resinous material produced by bees from various plant sources. The objective of this study was to characterize propolis samples of Apis mellifera from the Colombian Andean region, regarding the antimicrobial and physicochemical profiles. We used the technique of disk diffusion with ethanol extracts of propolis against Escherichia coli, Staphylococcus aureus and Candida albicans. The physicochemical characterization included percentage of solids content, wax, oxidation index and quantitative determination of phenolic and flavonoids compounds. All samples showed antibacterial activity, with inhibition zones between 8,0 and 12,0 mm for E. coli and between 8,3 and 23,5 mm for S. aureus. We did not observe any activity against C. albicans. The physical and chemical parameters cited above showed a variation from 2.72 to 9.17%, 0 to 2%, 3 to 51 s, 0.1 to 0.5 (w/w) and 0.02 to 0.95 (p/p), respectively. The antimicrobial profile observed, related to the physicochemical profile, suggests the need for further studies to determine the geographical and botanical origin of the samples studied.