India has one of the world’s richest flora with about 120 families of plants comprising 1,30,000 species and about 119 secondary plant metabolites are used globally as drugs. Hemidesmusindicus, an Indian herb which has potential of medical importance, which is supported by the present study . In this study bioactive compound2-Hydroxy-4-methoxybenzaldehyde (2H4MB) was isolated which has shown potential of antibacterial and antifungal activities.2-Hydroxy-4-methoxybenzaldehyde (2H4MB) was the major chemical entity (99.41%) amongst 10 identified compounds. Due to bioactive property Hemidesmus indicus could be utilized to control infectious diseases caused by the test pathogens and Candida albicans in the human system. The in vitro antibacterial activity of 3 different extracts (Haxane, Methanol and Aqueous) and Bioactive compound 2-Hydroxy-4-methoxybenzaldehyde (2H4MB) as well as MIC values of isolated compound (2H4MB) were performed. The most active extract was found to be the hexane extract showing the maximum zone of inhibition of 22 mm against Staphylococcus aureus.