Abstract: The microwave-assisted extraction (MAE) of antioxidant compounds from mango seeds was optimized using response surface methodology. The important factor influencing antioxidant activity was identified by a Plackett–Burman design, which determined three factors as follows: significant solid liquid ratio (X1), temperature (X2) and extraction cycle (X3), respectively. A Box–Behnken design (BBD) was applied to optimize the significant factors. The optimized conditions are X1 of 1/60 g/mL, X2 of 75 °C, and X3 of 2 cycles. Under these conditions, the antioxidant activity is 1738.2 mg trolox/g. IC50 (0.078 mg/mL), which indicates the phenolic compounds possess an antioxidant activity greater than the commercial antioxidants. RP-HPLC-ESI-MS analysis reveals that ethyl gallate, Penta-O-galloyl-glucoside (PGG) and Rhamnetin-3-[6″-2-butenoil-hexoside] are the major composition in the mango seed extracts. This study revealed that MAE is an efficient method for extracting compounds with relevant biological properties.