Allium hirtifolium belongs to Alliaceae family is traditionally used as flavoring agent and as natural remedy for treatment of infectious diseases. In this study, we analyzed A. hirtifolium essential oil by GC and GC-MS; the antioxidant and antimicrobial activity of A. hirtifolium essential oil were evaluated in vitro condition. 5-chloroorcylaldehyde (55.1%), methyl methylthiomethyl disulfide (24.6%) were the major components of oil. The antioxidant activity of oil (IC50 = 1.59%) were compared with BHT (IC50 = 0.002%). In beta-carotene test, the oil ( 1.59%) showed 32.3% inhibition, while 0.002% BHT showed 99.3% inhibition. The MIC and MLC values of A. hirtifolium oil were in the ranges of 0.06-2 and 0.25-2 mu l/ml, respectively. Aspergillus flavus was more sensitive to oil than that of other microorganism. A. hirtifolium oil exhibited high antimicrobial activity against bacteria, yeast and fungi. So, A. hirtifolium can be added to foods as antimicrobial or antioxidant agent in addition of its flavoring effect.