高选择性和高吸附容量混合模式色谱固定相的可控制备及应用研究

How to prepare stationary phase with selectivity and high coulmn capacity is an important scietific issue in separation science, which has been exploring for a long time. Grafting functional groups with high density onto solid phase matrix is the key to prepare high-capacity and high-selectivity stationary phase. Based on our previous work, this project selects two or three functional monomers and aims to graft functional block polymers and coplomers at high density onto silica beads by atom transfer radical polymerization to prepare novel mixed-mode stationary phase with high-capacity and high-selectivity by enhancing loaded-capacity of functional groups and adjusting the type of functional group and its density on the surface of stationary phase. To further improve the selectivity, the charge and polar of stationary phase will be adjusted by changing the ratio of two monomers in polymers. Using standard proteins, peptides and Chinese medicine components as solutes, the relationships between the column capacity and selectivity with the grafting degree of polymer and the proportion of monomer ratio will be investigated. The project can provide theoretical guidance and experimental support for the preparation of the stationary phase with high-selectivity and high-capacity. By analyzing small molecular and purifying antibody and protein tergets in biological samples, the application prospects of the prepared stationary phases will be elucidated in separation of complex samples.

如何制备高选择性和高吸附容量的色谱固定相是分离科学中长期探索的重要科学问题。将不同功能团高密度地接枝在基质表面是制备高负载量和高选择性固定相的关键。在前期工作基础上，本项目选择两种或三种功能单体，采用原子转移自由基聚合（ATRP）技术，在硅胶微球表面高密度地修饰功能嵌段聚合物和共聚物，通过提高功能团负载量和调控功能团种类和密度，制备出高选择性和高负载量的混合模式色谱固定相；通过调节嵌段聚合物和混聚物中两种单体的比例，调控固定相表面电荷和极性，提高选择性；以蛋白质、多肽、中药组分等标准品为溶质，探索柱容量和分离选择性与功能单体接枝量、聚合物链段中两种单体比例之间的关系，为高选择性和高负载量固定相的制备提供理论指导；以生物样品中小分子分析、抗体和蛋白药物纯化为例，阐明这两类固定相在复杂样品分析、目标物分离纯化中的应用价值。

针对分离科学领域内固定相吸附容量低，选择性不高的问题，本项目采用原子转移自由基聚合（ATRP）和光引发自由基聚合技术，将不同功能单体高密度地修饰在基质表面，通过提高功能团负载量和调控功能团种类，建立了选择性和高负载量的混合模式色谱固定相和吸附剂的制备方法。（1）在采用表面ATRP技术研究中，对邻羟基物质如核苷的分析，选择了丙烯酰氨基苯硼酸类单体；对食品和环境中的有害分子分析，选择了一系列疏水性/离子性单体或疏水/亲水单体进行ATRP聚合制备固定相和吸附剂。（2）在用光引发自由基聚合方法中，选择多功能基单体，以及可降解聚合物致孔剂进行聚合反应，再结合点击化学反应进行后修饰，制备适合不同分析对象的垂直孔整体柱。用色谱分离和固相分散萃取技术，证明了吸附容量对链长具有正相关性，选择性取决于高分子结构单元组成。阐明了以新材料为核心的分离富集技术具有选择性好、吸附剂用量少、富集能力强等优点，在生命分析、环境分析、生物大分子分离纯化等领域具有潜在的应用价值，同时所建立的吸附剂制备方法对于表面浸润、摩擦等表界面材料的结构设计具有一定的借鉴作用。