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This work is part of the valuation of essential oil of Mentha Rotundifolia harvested in the mountains of El Hamdania, in the Atlas Blideen (northern Algeria). The essential oil of Mentha rotundifolia extracted by hydro distillation have been analysed by means of Gas Chromatography GC-FID and GC-FID coupled to a Mass Spectrometer GC-MS in combination with retention indices. 34 constituents were identified accounting for 92 % of the essential oil. The main components were piperitenone oxide (30.37%)   and piperitone oxide (23.78%). The antimicrobial activity evaluated on seven microbial strains revealed sensitivity for the majority of the bacteria except pseudomonas aeruginosa. The antioxidant activity was determined according to the ability of the tested samples to scavenge the free radicals 2,2-diphenyl-1-picrylhydrazyl (DPPH*). The essential oil were slightly active 21.8% comparing with Quercetin (98.2%) of these oils can be attributed to the absence of some phenolic components, which may play an important role. The inhibitory effect of this essential oil on the development of bacteria and more particularly its effect on staphylococcus aureus sees glimpses of applications in the pharmaceutical industry as a natural antibiotic, and for other applications in the food and cosmetic industries.