Abstract: The column chromatographic fractions of chloroform (CHL1, CHL2 and CHL3) root extracts of Boscia angustifolia were screened for antibacterial activity and phytochemical properties. CHL1 fraction was significantly active (p<0.05) at 5 to 60 mg L-1 on Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Streptococcus pneumoniae with Minimum Inhibitory Concentration (MIC) of 0.63 to 1.25 mg L-1. The active fraction (CHL1) revealed the presence of only alkaloids with Retention factor (Rf) value of 0.36. The active antibacterial agent in the most potent fraction (CHL1) was isolated and identified by thin layer chromatography (TLC) and phytochemistry. The CHL2 and CHL3 fractions did not show inhibitory activity at 5 to 60 mg L-1. The antibacterial activity of root extract of Boscia angustifolia is due to a chloroform-extractable compound. The results support the ethnomedicinal use of root of Boscia angustifolia for the treatment of bacterial diseases.