The dried and powdered root of Rumex abyssinicus was extracted with increasing gradient of solvents (petroleum ether, chloroform, acetone and methanol) and screened for antibacterial activity on four pathogenic bacterial strains: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Salmonella thyphimurium. The acetone extract showed promissing antibacterial activity. Chromatographic separation of the acetone extract led to isolation of three compounds of which two of them (Betulone and oleic acid) were isolated for the first time from the plant. The structures of these compounds were charachterized with the help of spectroscopic methods (IR, NMR). Two compounds, RA-1 and RA-2, were identified to be betulone and oleic acid respectively based on spectroscopy data and literature reports, whereas the third compound RA-3 was only partially characterized. Ciprofloxacin was used as a reference drug during antibacterial assay.