Glycosides of the Citrus laurantifoia L. fruits have been extracted, then number of preliminary qualitative chemical tests were carried out on the extract. It was found that the extract consist of three glycoside compounds (G1, G2 & G3). Compound (G1) has been isolated from Citrus laurantifoia L. fruits by column chromatography using n-Butanol : Acetic acid : Water (4:1:5, upper layer) as eluent. The possible antibacterial activity of glycoside compound (G1) was assayed against the bacteria, Staphylococcus aureus, Streptococcus aureus, Pseudomonas aeruginosa and Escherichia coli using agar diffusion method. The results revealed that compound (G1) exhibited antibacterial activity against all the bacteria tested indicating that the presence compound had broad spectrum antibiotic effect.