Introduction: In recent years, people are more health-conscious and avoiding the use of synthetic drugs, which are not only harmful to them but also to the environment as they are manmade. So, there is an urgent need for naturally available biologically active compounds which are environment friendly and has tremendous health benefits for the welfare of human being. Trachyspermum Ammi, commonly known as ‘Ajwain,’ is a plant native to India. It has been shown to have antifungal, antioxidant, antimicrobial, antinociceptive, cytotoxic, hypolipidemic, antihypertensive, antispasmodic, broncho-dilating, antilithiasis, diuretic, abortifacient, antitussive, nematicidal, Anthelmintic and antifilarial properties. Furthermore, studies reveal the presence of a variety of phytochemical constituents, including carbohydrates, glycosides, saponins, phenolic compounds, volatile oil (thymol, -terpinene, para-cymene, and- and -pinene), protein, fat, fibre, and mineral matter containing calcium, phosphorous, iron, and nicotinic acid. Methods: TLC of essential oil of Trachyspermum ammi, was performed and revealed the presence of. Thymol. GC-FID & GC-MS analysis of essential oil shows the presence of 31 compounds. The most existing major compounds in essential oil were Thymol (33.40 %), γ-Terpinene (16.58 %), β-Pinene (13.82 %), 2-Pinene (6.02 %), 3-Cyclohexene-1-carboxaldehyde, 1,3,4-tri-methyl (5.35 %), p-Cymene (3.85 %), Linalool (2.89 %) and α-Thujene (2.44 %). Results: Antimicrobial activity of essential oil showed against Staphylococcus aureus and E. coli, it was proven by using cup well method by observing zone of inhibition to the microbial strains at different concentrations of essential oil. The minimum inhibitory concentration was also observed. Conclusion: It was confirmed that essential oil shows the antimicrobial activity as well as shows effect to fungal strains. In conclusion, the presence of numerous essential oil ingredients, and the results of experimental studies showed that Trachyspermum ammi essential oil possessed antioxidant, antibacterial, and antifungal action. Vulvovaginal candidiasis can be treated with this essential oil.