Aerial parts of Mercurialis annua L. were used in the present study, and fatty acid content in lipid extract was determined using GC-FID. The major fatty acids identified were αñlinolenic acid (20.3%), heptadecanoic acid (12.8%), palmitic acid (11.9%), pentadecanoic acid (11.7%), cis-10-pentadecanoic acid (11.2%), linoleic acid (7.7%), tridecanoic Acid (4.6%), stearic (4.4%), cis-11,14-eicosadienoic acid (3.8%), beside of minor fatty acids (palmitoleic acid, cis-13,16-docosadienoic acid, arachidic acid, behenic acid, cis-10heptadecenoic acid and myristic acid). Antioxidant properties of the extract were determined via DPPH radical scavenging, β-carotene bleaching assay and NO radical scavenging assay. The extract produced significant antioxidant activity in-vitro. The data shown here may broaden our knowledge on composition and antioxidant activity of lipid constituents from Mercurialis annua L.