Aim of the study: The aerial parts of Pamburus missionis (Wight) Swingle (Rutaceae) are traditionally used in the treatment of swelling, chronic rheumatism, paralysis, and puerperal diseases.The aim of this work was to investigate the chemical composition and antibacterial activity of essential oil of Pamburus missionis against different bacterial strains. Materials and methods: The essential oil was obtained from fresh leaves by hydro distillation method and its chemical composition was analyzed by GC and GC-MS. The antibacterial activity was determined by disc diffusion and micro broth dilution assay. Results: GC-MS analysis of the essential oil revealed the presence of 1-tridecanol (38.4%), n-hexadecanoic acid (16.1%), oxygenated monoterpenes (14.4%), monoterpene hydrocarbons (3.1%) and eugenol (1.9%) as the major components. The oil exhibited antibacterial activity at 100 and 450 μg against the test organisms with inhibition zones of 7-25 mm and minimal inhibitory concentrations values in the range of 10-100 and >100 mg/ml. Conclusion: The present study reveals the antibacterial potency of essential oil of Pamburus missionis. The use of Pamburus missionis in the treatment of various ailments and puerperal diseases can be attributed to its antibacterial property. © 2009 Elsevier Ireland Ltd. All rights reserved.