Sun Shaoai, Zhang Xiaoqiong, Han Qiang, Wan Wei, Ding Mingyu
Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
Talanta. 2016;149:187-193. doi: 10.1016/j.talanta.2015.11.042. Epub 2015 Nov 17.
Siliceous mesostructured cellular foam (MCF) with highly interconnected porous structure, ultralarge pore size and relatively uniform particle size (3-5μm) was prepared to achieve the mixed-mode and efficient separation of intact proteins. And molecular sieving effect for the first time played an important role in protein separation using mesoporous silica materials as HPLC stationary phase. The spherical silica particles were synthesized via hydrothermal method and the pore size was easily regulated by adding NH4F as well as altering the aging time. After aminopropyl derivatization, the chromatographic performance of functionalized mesoporous silica particles was investigated in comparison with those without modification and commercial NH2 column, and their mixed-mode retention mechanisms were investigated in detail. The superior separation performance for the retention of proteins was obtained on our home-made column in comparison with commercial NH2 column. The influences of aminopropyl derivatization and mobile phase composition on the column property were also investigated. Moreover, the home-made column showed similar performance for separation of polar anilines and neutral PAHs with the commercial column, owing to mixed-mode retention mechanisms including p-π stacking, electron interaction, hydrophobic effect, π-π EDA interaction and hydrogen bonding. All these results indicated that the aminopropyl modified MCF would be promising in the mixed-mode and efficient separation of biomolecules in addition with small molecules.
制备了具有高度互连多孔结构、超大孔径和相对均匀粒径（3-5μm）的硅质介孔泡沫材料（MCF），以实现完整蛋白质的混合模式高效分离。首次在以介孔二氧化硅材料作为高效液相色谱固定相的蛋白质分离中，分子筛分效应发挥了重要作用。通过水热法合成了球形二氧化硅颗粒，通过添加NH4F以及改变老化时间可轻松调节孔径。进行氨丙基衍生化后，将功能化介孔二氧化硅颗粒的色谱性能与未改性颗粒和市售NH2柱进行了比较，并详细研究了它们的混合模式保留机制。与市售NH2柱相比，我们自制的柱子在保留蛋白质方面具有优异的分离性能。还研究了氨丙基衍生化和流动相组成对柱子性能的影响。此外，由于包括p-π堆积、电子相互作用、疏水作用、π-π电子给体-受体相互作用和氢键在内的混合模式保留机制，自制柱在分离极性苯胺和中性多环芳烃方面表现出与市售柱相似的性能。所有这些结果表明，氨丙基改性的MCF在生物分子与小分子的混合模式高效分离方面具有广阔前景。
