Commiphora swynnertonii (Burtt) is traditionally used for medicinal purposes both in humans and animals in Tanzania. Previously, its leaves extract has been demonstrated antibacterial activity. However, there is limited information on identification of pure compounds responsible for its observed antibacterial activity. Therefore, this study was conducted to isolate and evaluate antibacterial activity of pure compounds isolated from its leaves extract. Colum chromatographic separation of its dichloromethane leaves extract afforded the isolation of two sterols namely, Cholest-7-en-3?-ol (1) and ?-sitosterol (2). Their structures were deduced by using NMR data experimentally obtained and comparison with spectral data available in literatures. The sterols were evaluated for activity against four nosocomial infectious bacteria namely; Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa to determine Minimum Inhibitory Concentrations (MIC) by broth microdilution method. Cholest-7-en-3?-ol (1) demonstrated strong activity against S. aureus (MIC= 0.5 mg/ml) and weak activities (MIC >2mg/ml) against other bacteria. ?-sitosterol (2) exhibited weak activities (MIC >2mg/ml) against all tested bacteria. Therefore, presence of antibacterial active compound(s) such as Cholest-7-en-3?-ol (1) among others in the leaves of C. swynnertonii can justify its earlier reported antibacterial activity and further confirms the claimed traditional uses of this plant in the treatment of bacterial infections.