高速逆流色谱法分离制备南极磷虾虾粉抗氧化肽  

Separation and preparation of Euphausia superba antioxidant peptides by high speed countercurrent chromatography

作　　者：李芙蓉 施文正[1,2,3] 李燕 Li Furong;SHI Wenzheng;LI Yan(College of Food Science,Shanghai Ocean University,Shanghai 201306,China;Engineering Research Center for Processing and Storage of Shanghai Seawater Products,Shanghai 201306,China;National Freshuater Aquatic Products Processing Technology R&D Center,Shanghai 201306.China)

机构地区：[1]上海海洋大学食品学院,上海201306 [2]上海水产品加工及贮藏工程技术研究中心,上海201306 [3]国家淡水水产品加工技术研发分中心,上海201306

出　　处：《上海海洋大学学报》2019年第2期305-312,共8页Journal of Shanghai Ocean University

基　　金：国家科技支撑计划(2015BAD17B02)

摘　　要：采用复合蛋白酶酶解南极磷虾得到虾粉多肽,经超滤分离和高速逆流色谱技术进行分离纯化并检验其体外抗氧化活性。结果表明:经超滤分离后的多肽在分子量范围为5～10 ku时多肽抗氧化活性最好,当浓度为10 mg/m L时,其超氧阴离子自由基清除率达到21. 28%,DPPH自由基清除率为86. 52%,对羟基自由基清除率为53. 49%,Fe^(2+)螯合率为85. 35%。对分子量范围在5～10 ku的多肽进行高速逆流色谱分析,采用体积比为氯仿∶丁醇∶甲醇∶水=4∶0. 75∶3∶2作为两相溶剂体系,下相作固定相,上相作流动相;反转,转速900 r/min,恒温水浴温度为25℃,在流动相的流速1. 5 m L/min,进样浓度20 mg/m L的条件下分离得到5个组分。经测定:当浓度为5 mg/m L时,P3组分清除超氧阴离子自由基的能力和Fe^(2+)螯合能力均显著高于其余4个组分,分别为37. 18%和92. 33%,P2组分的DPPH自由基清除率最高,为90. 33%,P5组分的羟基自由基清除率最高,为70. 11%。与未经HSCCC分离的对照组相比,抗氧化活性均得到明显增强。The compound peptide was used to digest the Euphausia superba to obtain the shrimp powder polypeptide,which was separated and purified by ultrafiltration separation and high-speed countercurrent chromatography to test its antioxidant activity in vitro.The results showed that the peptides isolated by ultrafiltration had the best antioxidant activity when the molecular weight ranged from 5 to 10 ku.When the concentration was 10 mg/mL,the superoxide anion radical scavenging rate reached 21.28%,and the DPPH free radical scavenging rate was 86.52%,the hydroxyl radical scavenging rate was 53.49%,and the Fe^2+chelation rate was 85.35%.High-speed countercurrent chromatography was carried out on peptides with a molecular weight ranging from 5 to 10 ku,using chloroform-butanol-methanol-water=4:0.75:3:2 as the two-phase solvent system,the lower phase as the stationary phase and the upper phase as the mobile phase;As for inversion,the speed was 900 r/min,the constant temperature water bath temperature was 25℃,the flow rate of the mobile phase was 1.5 mL/min,and the five components were separated under the injection concentration of 20 mg/mL.It was determined that when the concentration was 5 mg/mL,the ability of P3 component to scavenge superoxide anion radicals and Fe2+chelation ability were significantly higher than the other four components,which were 37.18%and 92.33%,respectively.The free radical scavenging rate of P2 was the highest,which was 90.33%,and the P5 component had the highest hydroxyl radical scavenging rate of 70.11%.Antioxidant activity was significantly enhanced compared to the control group not isolated by HSCCC.

关 键 词：南极磷虾 多肽 酶解 高速逆流色谱 抗氧化 

分 类 号：TS254[轻工技术与工程—水产品加工及贮藏工程]