固定化白桦酸亲和整体柱的制备方法及其应用  

Preparation and application of a betulinic acid functionalized affinity monolithic column

作　　者：黄小玲 杨鸿雁 单紫轩 袁英才 汪锦才 赖亮 刘正 郭嘉亮[1,2] HUANG Xiao-ling;YANG Hong-yan;SHAN Zi-xuan;YUAN Ying-cai;WANG Jin-cai;LAI Liang;LIU Zheng;GUO Jia-liang(Pharmacy College,Jinan University,Guangzhou 510632,China;Foshan University,Foshan 528000,China)

机构地区：[1]暨南大学药学院,广州510632 [2]佛山科学技术学院,佛山528000

出　　处：《天然产物研究与开发》2023年第2期302-309,290,共9页Natural Product Research and Development

基　　金：国家自然科学基金(81872832,82173781);广东省基础与应用基础研究基金自然科学基金(2019A1515010806);广东省教育厅重点领域专项(2020ZDZX2057)。

摘　　要：探究固定化天然产物白桦酸衍生物制备整体色谱柱的方法及应用。以天然活性小分子白桦酸衍生物3,23-O-二乙酰基-28-白桦酸胺乙基酰胺(EDA-BA)为配体,甲基丙烯酸缩水甘油酯(GMA)为单体,乙二醇二甲基丙烯酸酯(EDMA)为交联剂,正丙醇和1,4-丁二醇为生孔剂,偶氮二异丁腈(AIBN)为引发剂,通过原位聚合反应一锅法制备而成固定化白桦酸的整体柱poly(GMA-EDA-BA-co-EDMA);通过元素分析、扫描电镜、微径液相等手段对整体柱进行表征。液相色谱的条件下用于蛋白质的分离分析研究,发现除了α-葡萄糖苷酶,其他四种蛋白质和酶(包括胰蛋白酶、牛血清白蛋白、人血清白蛋白以及细胞色素C)在该柱上无特异性吸附,提示了该整体柱具有一定的选择性。研究表明,“一锅法”可应用于固定化天然产物单体化合物分子的整体柱的制备。To investigate the preparation method and application of immobilized betulinic acid(BA)derivatives in monolithic chromatography columns.The monolithic column poly(GMA-EDA-BA-co-EDMA)was prepared with a one-pot in situ copolymerizations of the ligand ethylenediamine-betulinic acid(EDA-BA),the monomer glycidyl methacrylate(GMA),the cross-linker ethylene glycol dimethacrylate(EDMA),the porogen(n-propanol/1,4-butanediol),and initiator 2,2′-azobisisobutyronitrile(AIBN),under optimized polymerization conditions.The columns were characterized by elemental analysis,scanning electron microscopy,and micro-HPLC.The result showed that most of the tested proteins and enzymes,including trypsin,bovine albumin,human serum albumin,and cytochrome C,have no retention in this column apart fromα-glucosidase,which the resulted monolithic column had a selective affinity toα-glucosidase.It was shown that the"one-pot"method can be well applied to the preparation of monolithic columns for the immobilization of natural molecules.

关 键 词：天然产物 白桦酸 整体柱 液相色谱 一锅法制备 

分 类 号：R284.2[医药卫生—中药学] Q503[医药卫生—中医学]