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Gynura bicolor DC. (G. bicolor) is a popular vegetable consumed in many Asian countries. In this study, chemical profile and antioxidant activity of G. bicolor ethanolic extract were studied. The extraction conditions were optimized and the total phenolic content of G. bicolor was determined as 10.38 mg of gallic acid equivalents per gram of dry matter. Twenty three compounds were identified in G. bicolor by HPLC-Q-TOF-MS analysis. Among them, 14 phenolic compounds, including ninepolyphenols and five flavonoids were found. 3-O- and 5-O-p-coumaroylquinic acid were the most abundant phenolic compounds in the vegetable. The ethanolic extract of G. bicolor showed appreciable antioxidant activity as analyzed by radical scavenging and reduce power tests.