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The antioxidant and antiviral activities of Euphorbia thymifolia L. (Euphorbiaceae) were investigated in this study. The results showed that all of the fractions (MeOH, CHCl₃, EtOAc, n-butanol and water) and pure compounds (3-O-galloyl-4,6-(S)-HHDP-D-glucose, rugosin B and 1,3,4,6-tetra-O-galloyl-K-β-D-glucose)tested possessed antioxidant activities, with the exception of the organic aqueous fraction in the anti-lipid and anti-superoxide formation assays. The range of IC₅₀ of anti-lipid formation, anti-superoxide formation and free radical scavenging assays for all fractions and pure compounds were 2.81–7.63, 0.03–2.18 and 0.013–2.878 mg/ml, respectively. Electron spin resonance studies showed that water extract and pure compounds of E. thymifolia exhibited superoxide radical and hydroxyl radical scavenging activities. Besides antioxidant activities, 3-O-galloyl-4,6-(S)-HHDP-D-glucose and EtOAc fractionalso showed anti-HSV-2 activity. Thus, E. thymifolia was concluded to possess antioxidant and anti-HSV-2 activities.