基于大孔径吸附树脂的模拟移动床色谱分离桑叶黄酮及其生物活性测定  

Separation of Flavonoids from Mulberry Leaf by Simulated Moving Bed Chromatography Based on Macroporous Adsorption Resin and Determination of Their Bioactivity

作　　者：李长龙 王金正 刘曦 王丹丹 盛晟[1,2] 王俊[1,2] 吴福安[1,2] Li Changlong;Wang Jinzheng;Liu Xi;Wang Dandan;Sheng Sheng;Wang Jun;Wu Fu’an(College of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang Jiangsu 212018,China;Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang Jiangsu 212018,China)

机构地区：[1]江苏科技大学生物技术学院,江苏镇江212018 [2]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《蚕业科学》2019年第4期569-576,共8页ACTA SERICOLOGICA SINICA

基　　金：现代农业产业技术体系建设专项(No.CARS-18)

摘　　要：为了获得高活性的桑叶黄酮类化合物,建立了基于大孔径吸附树脂的模拟移动床色谱分离工艺,并对制备的桑叶黄酮提取物的体外抗氧化、抑制酪氨酸酶等生物活性进行测定。根据大孔径树脂的静态和动态的吸附与解吸特性,从5种树脂中选取NKA-9型作为分离填料,上样时的最佳流速为6 BV/h、体积为4 BV,洗脱时的最佳流速为6 BV/h、体积为2.5 BV。在此基础上,构建了4区6柱的模拟移动床色谱分离桑叶黄酮,获得优化工艺条件为上样流速0.6 mL/min、洗脱流速1.2 mL/min、转换周期200 s,在该条件下所得桑叶黄酮的最高质量浓度为1.96 mg/mL,产量为141.12 mg/(BV·h)。生物活性测定结果显示,模拟移动床色谱分离桑叶黄酮的酪氨酸酶抑制率提高了3.3倍,其半抑制浓度(IC50)为0.14 mg/mL;自由基清除率提高了3.1倍,其IC50为0.095 mg/mL。模拟移动床色谱分离所得的桑叶黄酮纯度较高,抗氧化和酪氨酸酶抑制活性均得到明显提高。In order to separate flavonoids with high bioactivity from mulberry leaf, simulated moving bed chromatography separation process based on macroporous adsorption resin was established in our study, and bioactivity like in vitro antioxidation and tyrosinase inhibition was determined. In accordance to the static and dynamic adsorption and desorption characteristics of macroporous resin, NKA-9 type was selected from 5 types of resin as the separation media. The optimal loading flow rate was 6 BV/h and the volume was 4 BV. The optimal elution flow rate was 6 BV/h and the volume was 2.5 BV. On this basis, simulated moving bed chromatography of 4 areas and 6 columns was constructed to separate flavonoids from mulberry leaf with the following optimized process conditions: loading flow rate was 0.6 mL/min, elution flow rate was 1.2 mL/min, and transition period was 200 s. Under such conditions, maximum concentration of flavonoids from mulberry leaves was 1.96 mg/mL, and the yield was 141.12 mg/(BV·h). Bioactivity test showed that tyrosinase inhibition rate of flavonoids separated from mulberry leaf by simulated moving bed chromatography increased by 3.3 times, whose half maximal inhibitory concentration(IC50) was 0.14 mg/mL. Free radical scavenging rate increased by 3.1 times, whose IC50 was 0.095 mg/mL. In conclusion, flavonoids separated from mulberry leaf by simulated moving bed chromatography were better in purity, antioxidation activity and tyrosinase inhibition activity.

关 键 词：桑叶 黄酮类化合物 大孔径树脂 模拟移动床 酪氨酸酶抑制活性 抗氧化活性 

分 类 号：R284.4[医药卫生—中药学]