Abstract: In order to obtain higher purity grass carp head phospholipids and analyze the antioxidant properties of the prepared grass carp head phospholipids,phospholipids were prepared from freeze-dried grass head powder by ethanol extraction. After determined its optimal process by the orthogonal test,the composition and content of phospholipids from grass carp head were analyzed by thin layer chromatography(TLC). The fatty acid composition of the phospholipids was analyzed by gas chromatography-mass spectrometry(GC-MS),and the antioxidant activity in vitro was determined. The results showed that,the optimal process for preparing phospholipids was:Ethanol concentration 80%,material-liquid ratio 1:6 g/mL,extraction temperature 65℃,and extraction time 3 h. Under these conditions,the purity of grass carp head phospholipids were 84.20%±0.65%,and the extraction rate were 3.03%±0.06%.The grass carp head phospholipids mainly contained phosphatidylcholine(PC),phosphatidylethanolamine(PE),sphingomyelin(SM),and lysophosphatidylcholine(LPC),among which PC had the highest contents(60.00%±0.26%),followed by PE(20.00%±0.17%). The fatty acids of grass carp head phospholipids were mainly composed of polyunsaturated fatty acid(PUFA,52.09%±0.59%). The PUFAs in phospholipids were mainly C20:5n-3(eicosapentaenoic acid,EPA)and C22:6n-3(docosahexaenoicacid,DHA). The results of antioxidant assay in vitro showed that the hydroxyl radical(·OH)scavenging ability and reducing power of grass carp head phospholipids were significantly(P<0.05)better than the commercial soybean phospholipids as comparation.