Vitexin-2"-O-rhamnoside (VOR) is one of components of flavonoid of the leaves of Crataegus pinnatifida Bge.var major N.E.Br.[1], which has been demonstrated to strongly inhibit DNA synthesis in MCF-7 human breast cancer cells [2].Recently, the high sensitive and selective HPLC and UPLC-ESI-MS/MS methods have been employed to investigate the pharmacokinetic of VOR in biological fluids[3-5].Malondialdehyde (MDA) is used as marker of oxidative injury, and the concentration varies of MDA in response to the protective effect of vitexin-2"-O-rhamnoside (VOR) on ECV304 cell induced by oxidative stress.For the determination of MDA in human endothelial cell, ECV304 cell induced by tert-butyl-hydroperoxide (TBHP) to investigate the antioxidation of VOR, a commonly used, convenient high-performance liquid chromatography (HPLC) method was developed.