漆酚-松香基高分子印迹柱分离天麻素及其构效关系研究

This project aims at the synthesis of molecular imprinted polymer with gastrodin as the template, urushiol as monomer, and modified rosin as cross-linker. This polymer then will be used for the preparation of imprinted chromatography column which can be installed in HPLC instrument to establish on-line identification and separation MIPs-HPLC system. This system can be employed to detect and separate gastrodin which is a kind of effective compoment of Chinese herbal medicine. To obtain molecular imprinted polymeric column with high performance, firstly, factors influencing the properties of molecular imprinted polymer will be investigated, these factors include template molecule, monomer, cross-linker, porogenic solvent, polymerization process and reaction time. Secondly, the retention factor and partition coefficient of the urushiol-rosin-based imprinted polymer chromatography column will be explored. Additionally, the separation conditions of gastrodin on MIPs-HPLC system will be discussed and optimized. Lastly, the structure-property relationship between gastrodin and polymer structure, degree of crosslinking, specific surface, bore diameter, poriness, will also be investigated. As a result of the modified rosin molecule is large framework crosslinker, can synthesis huge bore diameter polymers. The urushiol-rosin-based polymer imprinted chromatography column will possesses excellent mechanical strength and high pressure resistance. In addition, the starting materials which synthesis urushiol-rosin -based molecular imprinted polymer derived from the non-toxic, environmental friendly, and inexpensive renewable resources. This work will provide new method and theoretical basis for exploring new application of the urushiol and rosin, and separation of Chinese herbal medicine effective components.

本项目以天麻素为模板分子，以漆酚为单体，以改性松香为交联剂，经交联聚合成漆酚-松香基分子印迹聚合物，制备漆酚-松香基高分子印迹柱，建立MIPs-HPLC在线检测分离系统分离纯化天麻素药物。探索模板分子、单体、交联剂、溶剂、反应时间、聚合方法等因素对分子印迹聚合物性能的影响，筛选出能有效分离天麻素的漆酚-松香基高分子印迹柱；研究漆酚-松香基高分子印迹柱分离天麻素的印迹指数、分离系数及MIPs-HPLC系统分离天麻素的分离条件；研究漆酚-松香基印迹聚合物的结构、交联度、比表面、孔径、空隙率等性质与天麻素之间的选择性构效关系。改性松香的分子骨架大，能合成大孔网状印迹聚合物，以漆酚-松香基聚合物制备印迹色谱柱，其机械强度好，可以耐高压；另外，合成漆酚-松香基聚合物的原料来源于可再生资源，丰产价廉且无毒环保。因此，本项目的研究将为探索生漆和松香的应用领域以及分离纯化中草药有效成分提供新的方法和理论。

以甲基丙烯酸甲酯等为单体，以丙烯海松酸丙烯酸乙二醇酯等为交联剂，采用悬浮聚合方法，分别制备了Ru131、Ru232松香基高分子微球、酯化修饰的Ru131松香基高分子微球、及SiO2@松香基高分子微球，采用扫描电镜、透射电镜、激光粒度测定仪、比表面积及微孔物理吸附仪、热重分析仪对合成的微球进行了表征。结果表明：各种松香基高分子微球粒径均一，球形度好，比表面积大，表面有丰富的孔结构，在300℃以内热稳定性良好。. 用制备的松香基高分子微球填充的系列色谱柱分离天麻素及其衍生物，并探索了流动相种类及浓度、流速、柱温等色谱条件对分离效果的影响。结果表明：Ru232色谱柱分离Gastrodin与Phenolglucoside的分离度Rs为1.54、分离Gastrodin与Methylarbutin的分离度Rs为2.73；Ru131-Mt分离Gastrodin与Methylarbutin的分离度Rs为2.30；SiO2@R3色谱柱分离Gastrodin与Phenyl β-D-glucopyranoside的分离度为8.52、分离Gastrodin与4-Methoxyphenyl-β-D-glucopyranoside的分离度为14.7、分离Phenyl β-D-glucopyranoside与4-Methoxyphenyl-β-D-glucopyrano -side的分离度为7.14。通过拟合范特霍夫方程分别计算了各对竞争分子的ΔH、ΔS、ΔG等热力学数据，并对其分离机理进行了分析。. 分别用Ru232色谱柱和SiO2@R3色谱柱分离纯化天麻水提物，分离纯化后的天麻素纯度分别达到82%和86.4%，说明合成的这些松香基高分子填料能够有效的分离纯化天麻素，具备较好的应用前景。