Abstract: In order to effectively utilize and develop Moringa oleifera seeds, the glycosides in Moringa oleifera seeds were extracted by microwave-assisted method with ethanol as solvent. Based on the single factor experiment, the extraction process was optimized by response surface methodology. The antioxidant activities in vitro of each polar part of the optimized Moringa oleifera seeds extract were determined by salicylic acid method, ABTS method and FRAP method. The results showed that the optimal extraction conditions were as follows: Ethanol volume fraction 81%, solid-liquid ratio 1:30 g/mL, extraction time 20 min, microwave power 600 W. The order of antioxidant activity in vitro was ethyl acetate>petroleum ether > n-butanol > water. The results indicated that the microwave-assisted extraction of glycosides from Moringa oleifera seeds and the optimization of the extraction process of total glycosides from Moringa oleifera seeds by response surface methodology was feasible. In vitro, antioxidant activity tests showed that the polar parts of ethanol extracts of Moringa oleifera seeds had free radical scavenging ability, which laid a foundation for the further development of natural antioxidants.