生物色谱用于当归中活性成分筛选及分离研究

Study on Sieving and Separation of Active Components of Chinese Angelica by Biochromatography

应用分子生物色谱技术研究了筛选和分离当归中的活性成分。以甲基丙烯酸缩水甘油酯为功能单体,通过原位聚合得到了整体柱基质,用二乙胺活化后,以吸附蛋白的方法得到了以牛血清白蛋白(BSA)为固定相的分子生物色谱柱。当归样品经甲醇提取,0.45μm膜过滤后,以BSA分子生物色谱柱为分离柱,以含2%(体积分数)甲醇的Tris-盐酸缓冲溶液为流动相进行分离,用二极管阵列检测器检测。结果表明:活性组分在BSA上的保留机理是疏水作用、静电作用和特定的与空间结构相关的多种因素的共同作用结果。将BSA柱上的保留组分进行了气相色谱-质谱分析,鉴定了8种成分,包括当归中的主要可挥发性成分(藁本内酯)。

Molecular biochromatography(MBC)was applied to the study on sieving and separation of active components of Chinese angelica.The matrix of monolithic column was prepared with glycidylmethacrylate as functional monomer by in-situ copolymerization.After activating the functional group with diethylamine,bovine serum albumin(BSA)was absorbed onto the matrix of monolithic column and a MBC column with BSA as stationary phase was formed.The sample of Chinese angelica was extracted with methanol,and the extract was filtered on 0.45μm filtering membrane,the filtrate was separated on the MBC column,with Tris-HCl buffer solution(containing 2% by volume of methanol as mobile phase.DAD was adopted in the determination.It was found that the mechanism of retention of the active components on BSA was the combining action of a variety of factors,including hydrophobic and electrostatic actions as well as the spatial structure.The components retained on BSA column were analyzed by GC-MS,and 8components were identified,including the main volatile component of ligustilide.