ABSTRAK Telah dilakukan isolasi, identifikasi komponen kimia dan uji antibakteri minyak atsiri daun lampes (Ocimum sanctum L.) pada Staphylococcus aureus dan Eschericia coli. Minyak atsiri diperoleh dengan metode destilasi stahl dan identifikasi komponen kimia dengan analisis data GC-MS. Uji aktivitas antibakteri minyak atsiri dilakukan dengan metode difusi. Hasil isolasi minyak atsiri dengan destilasi stahl diperoleh kadar 0,41%. Identifikasi komponen kimia dengan analisis data GC-MS adalah α-pinen (2,55%), kamfen (0,73%), sabinen (0,12%), β-pinen (0,54%), endo-borneol (0,76%), α-kopaena (2,55%), β-elemen (11,55%), metil-eugenol (41,90%), trans-kariofilen (25,66%), α-humulen (1,24%), germakren-A (1,19%), germakren-D (9,95%) dan δ-kadinen (0,65%). Aktivitas antibakteri dapat menghambat bakteri Staphylococcus aureus dan Eschericia coli. Nilai Konsentrasi Hambat Minimal (KHM) untuk Staphylococcus aureus adalah 787,5 ppm dan Eschericia coli adalah 525,5 ppm. Kata kunci : Minyak atsiri, Ocimum sanctum L., isolasi, aktivitas antibakteri, Staphylococcus aureus, Eschericia coli ABSTRACT Isolation, chemical component identification and antibacterial activity of the essential oil of Ocimum sanctum L. on Staphylococcus aureus and Eschericia coli have been done. The essential oil obtained by stahl destillation of Ocimum sanctum L. was analized by GC-MS. Antibacterial activity of essential oil was recognized by disc diffusion methods. The essential oil yield by stahl destillation is 0,41%. Compounds of essential oil identified by GC-MS are α-pinene (2,55%), camphene (0,73%), sabinene (0,12%), β-pinene (0,54%), endo-borneol (0,76%), α-copaena (2,55%), β-elemene (11,55%), methyl-eugenol (41,90%), trans-caryophyllene (25,66%), α-humulene (1,24%), germacrene-A (1,19%), germacrene-D (9,95%) dan δ-cadinene (0,65%). Antimicrobial activity of the essential oil was investigated against Staphylococcus aureus dan Eschericia coli. The Minimum Inhibitory Concentration (MIC) values for Staphylococcus aureus was determined at 787,5 ppm dan for Eschericia coli at 525,5 ppm. Keyword : Essential oil, Ocimum sanctum L., isolation, antibacteri activity, Staphylococcus aureus, Eschericia coli