LC Determination of Malondialdehyde Concentrations in the Human Umbilical Vein Endothelial Cell Culture Medium: Application to the Antioxidant Effect of Vitexin-2?O-rhamnoside

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• 作者：Xixiang Ying ; Haibo Li ; Zhili Xiong ; Zhaoshu Sun ; Shuang Cai ; Wenliang Zhu ; Yujin Bi and Famei Li
• 关键词：Column liquid chromatography ; Human umbilical vein endothelial cell ; Antioxidation ; Malondialdehyde ; Vitexin ; 2″ ; O ; rhamnoside
• 刊名：Chromatographia
• 出版年：2008
• 出版时间：May, 2008
• 年：2008
• 卷：67
• 期：9-10
• 页码：679-686
• 全文大小：482.9 KB

文摘

Vitexin-2″-O-rhamnoside (VOR) presents the leaves of Crataegus pinnatifida Bge. var. major which plays a role in preventing human pathologies related to oxidative stress, but as the principal component in the leaves, little attention has been devoted to its study of antioxidation of VOR. A simple, rapid and sensitive high-performance liquid chromatography method was developed to determine malondialdehyde (MDA) in ECV304 cell culture medium induced by tert-butyl-hydroperoxide (TBHP). The preparation of analyzed samples involved a one-step derivatization with thiobarbituric acid (TBA). The separation was achieved using the Synergi Hydro-RP, a polar endcapped C₁₈ column (250 ¡Á 4.6 mm, 4 μm pore size) with a linear gradient elution of acetonitrile and 10 mmol L−1 of ammonium acetate aqueous solution (pH 6.8). The calibration curve was linear over the ranges 0.0125–1.25 μmol L−1 MDA (r = 0.9962). The lower limit of quantification of MDA was 0.0125 μmol L−1. Relative standard deviations of intra-day and inter-day precision were less than 5.2 and 4.4 % . The method with high recovery (95.4 ¡À 1.3 % ) was successfully applied to the investigation of antioxidation of VOR by determining MDA in ECV304 cell culture medium.