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Paeonia daurica subsp. macrophylla (Paeoniacea) is a perennial plant growing in Iran. In Persian traditional medicine, Paeonia officinalis [Ood-e-Saleeb] have been used for treatment of some diseases especially for epilepsy and brain disorders. Based on initial phytochemical evaluations of Paeonia spp. roots, steroidal and terpenoid compounds were identified. Hydro distilled essential oil of the roots of P. daurica subsp. macrophylla collected from north of Iran, was investigated using gas chromatography-mass spectrometry (GC-MS). Altogether, five constituents, forming 96.02% of the total oil composition were identified. Major constituents of the essential oil were salicylaldehyde (20.32%), beta-pinene-oxide (13.35%) and thymol acetate (61.12%). Antioxidant effect of the ethanolic (80%) extract was measured using DPPH free radicals. According to the results, IC50 of the extract was 25.2 µg/ml, which can be assessed as strong antioxidant effect using standard Antioxidant Activity Index.