Gas chromatography mass spectrometry (GC-MS) was employed to determine the chemical composition of essential oil obtained from Eucalyptus globulus and Trachyspermum ammi. The essential oil obtained from leaves of Eucalyptus globulus and from seeds of Trachyspermum ammi by Clevenger apparatus. Chemical composition of the essential oils of E. globulus and T. ammi was analyzed by Gas chromatography – Mass spectrometry. Four main compounds in E. globulus were identified as Eucalyptol, Methyl – salicylate, Thymol, Para- cymene. 1, 8-cineole was found to be the main compound of E. globulus essential oil. Thymol, gammaterpinene, para- cymene, beta- pinene were the main compounds of T. ammi oil and traces amount of beta myrcene, alpha- thujne, alphapinene, alpha- terpinene, carvacrol compounds of T. ammi were identified by GC-MS. Thymol was the main compound of T. ammi oil. The antifungal activity of E. globulus and T. ammi oils were screened against Trichophyton mentagrophytes and Epidermatophyton floccosum by using disc diffusion and modified micro dilution method. T. ammi oil shows highest inhibition zone in comparison to E. globulus oils and references antibiotics i.e. Clotrimazole and Ketoconazole.