氨基酸离子液体功能化多作用机制色谱材料制备及蛋白质分离性能研究

The extreme complexity of protein samples renders the separation or fractionation step is crucial for further in-depth study of proteins. However, the existing chromatographic techniques are still difficult to resolve the problems of low separation efficiency and poor selectivity, especially for the complex protein samples. In this project, with the combination of theoretical and computational chemistry, a variety of imidazolium-based amino acid ionic liquids, possession of different hydrophilicity&hydrophobicity, acidity&basicity or aromaticity, will be firstly designed and synthesized. Furthermore, with the prepared polyhedral oligomeric silsesquioxane (POSS) hybrid monolithic materials or POSS modified silica (POSS@SiO2) core-shell particles as the matrixes, a series of novel mixed-mode chromatographic stationary phases with multiple interaction mechanisms will be prepared and further used for separation of complex proteins or specific proteins with high efficiency and selectivity. Moreover, the separation mechanisms and retention characteristics of the developed staionary phases will be further studied. This project could provide novel chromatographic stationary phases for protein separation with high efficiency, which is of great benefit to subsequent mass spectrometry based identification of proteins. Simultaneously, it could assist the design of more efficient chromatographic stationary phases in the future work.

蛋白质样品的极端复杂性使得分离和分级步骤对其深入地研究至关重要。本项目拟针对复杂蛋白质样品色谱分离分析中存在的分离效率低、分离选择性差的问题，通过结合理论化学计算辅助筛选，设计并合成多种具有不同亲疏水性、酸碱性或芳香性的咪唑基氨基酸离子液体，作为色谱功能化试剂，并以合成的具有优良特性的笼型聚倍半硅氧烷（POSS）杂化毛细管整体材料或POSS修饰二氧化硅（POSS@SiO2）的核壳型颗粒材料为基质，制备一系列具有多分离作用机制的新型混合模式色谱固定相，并深入研究其分离机制和分离特性，实现复杂蛋白质样品及特定类型蛋白质样品的高效、高选择性分离分析。该项目的研究成果可为蛋白质的高效分离分析提供新型色谱材料，从而为蛋白质的高效质谱鉴定在分离阶段提供强有力的保障；同时，可为后续进一步设计新型高效的色谱材料提供理论指导。

蛋白质样品的极端复杂性使得分离和分级步骤对其深入地研究至关重要。本项目针对复杂蛋白质样品色谱分离分析中存在的分离效率低、分离选择性差的问题，以合成的笼型聚倍半硅氧烷（POSS）杂化毛细管整体材料为基质，合成了5种氨基酸—离子液体修饰的杂化毛细管整体柱和2种离子液体功能化杂化毛细管整体柱，可实现反相/离子交换、反相/亲水等多模式色谱分离分析。所开发的新型固定相材料实现了蛋白质消解肽段、模式糖蛋白质和非糖蛋白质混合物、表达的TARG1蛋白质样品及卵清蛋白质样品的高效分离和纯化。优化了POSS@SiO2基质材料的合成方法，并以合成的POSS@SiO2或SiO2为基质材料，合成了2种氨基酸—离子液体、离子液体修饰的填充色谱分离材料，可实现反相/亲水/离子交换、反相/亲水混合模式色谱分离分析。此外，进一步拓展并制备了超疏水CMPs@SiO2、MOFs@SiO2核壳型填充色谱固定相和基于二季戊四醇戊-/己-丙烯酸的高交联杂化毛细管整体柱，并研究了其保留和分离分析特性。本项目所开发的新型固定相材料有望在复杂蛋白质样品的分离分级方面发挥重要的作用。在本项目的支持下，在Journal of Chromatography A、ACS Applied Materials & Interfaces、Microchimica Acta、Analytica Chimica Acta等期刊发表研究论文8篇，在TrAC-Trends in Analytical Chemistry撰写关于整体柱研究进展的综述论文2篇，申请相关发明专利3项。项目组成员参加国际国内学术会议4次，发表会议论文7篇。培养硕士研究生7人。