The anthocyanins in the fruits of Rhodomyrtus tomentosa (ACN) were extracted by 1% TFA in methanol, and then purified by X-5 resin column and C₁₈ (SPE) cartridges. The purified anthocyanin extract (ART) from the fruits of R. tomentosa showed strong antioxidant activities, including DPPH radical-scavenging capacity, ABTS radical scavenging capacity, reducing power and oxygen radical absorbance capacity (ORAC). The purified anthocyanin extract was analyzed by high performance liquid chromatography (HPLC). The major anthocyanins were purified by semi-preparative HPLC and Sephadex LH-20 column chromatography, and were identified as cyanidin-3-O-glucoside, peonidin-3-O-glucoside, malvidin-3-O-glucoside, petunidin-3-O-glucoside, delphinidin-3-O-glucoside and pelargonidin-3-glucoside by HPLC–ESI/MS and nuclear magnetic resonance spectroscopy (NMR). Cyanidin-3-O-glucoside was considered as the most abundant anthocyanin, which was 29.4mg/100g dry weight of R. tomentosa fruits. Additionally, all the major anthocyanins were identified from R. tomentosa fruit for the first time.