Euphorbia hirta L. has been proved a medicinally important plant. This study aimed to investigate the antioxidant capacities of fractioned extracts and some isolated compounds from Euphorbia hirta L. on pancreatic β-cells MIN6 exposed to hydrogen peroxide-oxidative stress conditions. Four pure compounds were isolated from ethyl acetate and methanol extracts and identified as quercitrin (1), luteolin (2), quercetin (3) and caffeic acid (4). The results showed that the ethyl acetate extract as well as quercetin possess strong protective effects with the cell viability of 81% and 82% at the dose of 0.1 mg/mL. This proves that E hirta L. is a potentially natural antioxidative plant and worth testing for further pharmacological potential in the treatment of oxidative stress which may be related to neurodegenerative diseases.