亚2微米硅胶基质多模式复合固定相的制备与分离机理研究

Ultra performance liquid chromatography systems utilize sub -2-micron porous particles as column materials and have demonstrated significant improvement in speed, sensitivity and resolution compared to conventional HPLC. UPLC has already become one of the most commonly used analytical tools in chemistry, food hygiene, drug testing, life science, and many other areas. However, there are still many issues existed in UPLC technology, and the core technology is mainly mastered by foreign researchers. For example, it was difficult to prepare monodisperse sub-2-micron silica particles directly, and stationary phases for UPLC was too simple to make full and deeply use of it. Besides, chromatographic retention mechanism was still not clear at present. In order to break up the foreign technical monopoly and develop domestic UPLC technology, this study will focus on preparation and multi-modification of sub-2-micron silica particles, and will study retention mechanism of the stationary phase. Firstly, since we have prepared the mesoporous 5-micron silica particles, monodisperse sub-2-micron silica particles will be directly prepared by capillary auxiliary granulation method and sol-gel coupled with PICA method. Secondly, sub-2-micron mesoporous silica particles will be multi-modificated by different group in order to prepare polar-embedded stationary phase and HILIC/Reversed stationary phase. In addition, based on quantum mechanics and molecular mechanics principle, an attempt is made to simulate the separation process in UPLC column and study the retention mechanism of the stationary phase. Then, a series of chromatographic tests will be conducted to determine the suitability of the new stationary phase for various UPLC applications. Finally, a series of separation methods for different samples on UPLC system will be established. This study not only give suggest to the design and preparation of stationary phase for special samples, but also provide basis for the choice of stationary phase for special samples. Moreover, foundation will be laid for industrialization of stationary phase for UPLC in our country.

基于亚2微米填料的超高效液相色谱（UPLC），以其高速度、高灵敏度、高分离度的特点，在食品安全、药物分析、生命科学等应用领域已得到快速推广。面对国外在该类研究中的高端技术垄断，针对目前国内亚2微米硅胶制备单分散性差、键合相单一、机理研究缺乏理论支持等现状，本项目在前期系统研究制备单分散5微米硅胶固定相的基础上，引入毛细管造粒，开展直接合成"单分散"亚2微米高效硅胶固定相的系统研究；在所制备的硅胶表面，进行多模式不同位点的复合改性，制备内嵌极性官能团、HILIC/反相混合模式固定相；通过量子力学和分子力学计算，模拟固定相表面样品的色谱行为，研究分离机理，评价固定相分离能效；将制备的亚2微米硅胶复合固定相应用于UPLC系统，针对实际样品进行分析方法发展。通过研究，力争为色谱用固定相"靶向"设计、筛选制备具有"特异性"机理的填料提供依据，为我国超高效液相色谱填料产业的技术研究提供基础数据。

超高效液相色谱技术，以其高速度、高灵敏度、高分离度的特点，在各应用领域得到快速推广。作为其分离关键核心的色谱填料，尤其是复合模式的新型填料，几乎被国外所垄断。.项目组系统研究了超纯硅胶的传统制备方法，一次催化溶胶凝胶法、二次催化溶胶凝胶法、模板法、聚合诱导胶体凝聚法(PICA)，考察研究了不同原料性能及反应条件等对微球粒度、形貌、孔结构等物理参数以及色谱分离性能的影响。在此基础上，发展了一种自制溶胶-聚合诱导胶体凝聚法，可直接制备5µm、亚2µm单分散性硅胶微球。.在制备硅胶微球的基础上，以三聚氯氰为反应物，γ-氨丙基三乙氧基硅烷为偶联剂，采用固液相表面连续法，制备了一种5µm内嵌三嗪环酰胺的复合模式的固定相，并采用傅里叶红外光谱和元素分析法对其进行表征。结果表明，配体已成功地键合到硅胶表面，所制备的固定相具有较高的碳载量。采用优化的合成方法，进行多次重复性试验，获得了良好的重复性结果。将该合成方法发展到亚2µm硅胶基质填料上，成功制备1.8µm内嵌三嗪环酰胺的复合模式固定相。.采用Tanaka等提出的方法，评价了所制备固定相装填色谱柱的基本性能，并与常规的C18色谱柱进行了对比。结果表明，该固定相具有较高的立体选择性和较低的氢键结合能力、总离子交换能力。