Optimization of total flavonoid compound (TFC) extraction from Gynura medica leaf was investigated using response surface methodology (RSM) in this paper. The conditions investigated were 30–60% (v/v) ethanol concentration (X1), 85–95 °C extraction temperature (X2) and 30–50 (v/w) liquid-to-solid ratio (X3). Statistical analysis of the experiments indicated that temperature and liquid-to-solid ratio significantly affected TFC extraction (p < 0.01). The Box-Behnken experiment design showed that polynomial regression models were in good agreement with the experimental results, with the coefficients of determination of 0.9325 for TFC yield. The optimal conditions for maximum TFC yield were 55% ethanol, 92 °C and 50 (v/w) liquid-to-solid ratio with a 30 min extraction time. Extracts from these conditions showed a moderate antioxidant value of 54.78 μmol quercetin/g dry material (DM), 137.3 μmol trolox/g DM for 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 108.21 μmol quercetin/g DM, 242.31 μmol trolox/g DM for 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS+), respectively. HPLC-DAD-MS analysis showed that kaempferol-3-O-glucoside was the principal flavonoid compound in Gynura medica leaf.