毛细管电泳法测定过氧化氢酶与天然抗氧化剂协同清除羟自由基作用  

Determination of Synergistic Effect of Catalase and Natural Antioxidants for Scavenging Hydroxyl Radical by Capillary Electrophoresis

作　　者：张丽[1] 陈冠华[1] 方柔[1] 伊聆晓[1] 邵钰秀[1] 肖晶[1] 余云娟[1] 陶香香[1] 

机构地区：[1]江苏大学食品与生物工程学院,镇江212013

出　　处：《分析化学》2013年第10期1571-1576,共6页Chinese Journal of Analytical Chemistry

基　　金：教育部高等学校博士学科点专项科研基金(No.20093227110010);江苏大学科研基金(No.08JDG001);江苏高校优势学科建设工程资助项目

摘　　要：在抗坏血酸／Fe^2＋羟自由基产生体系中，以苯甲酸为羟自由基捕获剂，建立了羟自由基的毛细管电泳分析方法。优化了背景缓冲液的pH值、电解质浓度、表面活性剂浓度及分离电压。在最优的电泳条件下，研究了羟自由基产生体系中各底物浓度和反应时间对产物生成量的影响。本方法可在15min内实现体系中的各物质的分离；产物间羟基苯甲酸的线性范围是5X10^-6一1×10^-3mol／L，检出限为2．8X10^-7mol／L。将本方法用于血清基质中过氧化氢酶、原花青素和绿原酸对羟自由基IC50的测定，并研究了过氧化氢酶与原花青素及与绿原酸的协同清除羟自由基作用。结果表明，过氧化氢酶与原花青素及与绿原酸均存在协同清除羟自由基作用，在过氧化氢酶与原花青素的协同作用间还存在量效关系。Capillary electrophoresis was employed to determine hydroxyl radicals trapped by benzoic acid in ascorhic acid/Fe2 ＋ system. The pH values of the background buffer, electrolyte concentration, surfactant concentration and separation voltage were optimized. The impacts of substrate concentration and reaction time on the production of hydroxyl radicals were invenstigated using the optimized eleetrophoresis conditions. All the substances in the system were separated within 15 min; the linear range of reaction product （m-hydroxybenzoic acid） , was 5 x 10^-6 mol/L to 1 x 10-3 mol/L, and the detection limit was 2.8× 10-7 mol/L. This method was applied to determine the ability of catalase, proanthocyanidins （PACs） and chlorogenie acid （CGA） for scavenging hydroxyl radical individually （IC50） and combined with each other in the serum medium. The result showed that catalase and PACs, catalase and CGA had synergistic effect in scavenging hydroxyl radical and this effect to eatalase and PACs was also dose-dependent.

关 键 词：过氧化氢酶 原花青素 绿原酸 羟自由基 协同抗氧化 毛细管电泳 

分 类 号：TS201.2[轻工技术与工程—食品科学] R285[轻工技术与工程—食品科学与工程]