Silica-based polypeptide-monolithic stationary phase for hydrophilic chromatography and chiral separation.
Silica-based polypeptide-monolithic stationary phase for hydrophilic chromatography and chiral separation.
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DOI：
10.1016/j.chroma.2016.04.014
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发表时间：
2016-05
期刊：
影响因子：
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通讯作者：
Licong Zhao;Limin Yang;Qiuquan Wang
中科院分区：
文献类型：
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作者：
Licong Zhao;Limin Yang;Qiuquan Wang
设计并合成了谷胱甘肽（GSH）、生长抑素乙酸酯（ST）和卵类粘蛋白（OV）功能化硅胶整体固定相，用于毛细管电色谱（CEC）的HILIC和手性分离。通过γ-缩水甘油氧基丙基三甲氧基硅烷（GPTMS）中缩水甘油基与GSH、ST和OV的氨基之间的环氧开环反应，以及四甲氧基硅烷与GPTMS的缩聚共聚反应，将GSH、ST和OV共价结合到SiO2骨架中。不仅可以通过移动的相的pH值来控制所制备的GSH-、ST-和OV-硅胶杂化整体固定相上的电渗流方向和电渗流大小，而且还观察到典型的HILIC行为，使得核苷酸和HPLC肽标准混合物可以在CEC期间使用不含任何乙腈的水性移动的相进行基线分离。此外，所制备的整体柱对l-氨基酸具有手性分离能力。OV-硅胶杂化整体柱对谷氨酸（Glu）、酪氨酸（Tyr）和组氨酸（His）的分离度分别为1.07、1.57和1.06。重要的是，GSH-硅胶杂化整体柱的手性分离能力可以显着增强时，使用金纳米粒子（AuNP），以制造AuNP介导的GSH-AuNP-GSH-硅胶杂化整体柱。Dl-Glu、Dl-Tyr和Dl-His的RofD分别为1.19、1.60和2.03。将该整体柱应用于药物对映体的分离，实现了所有四种R/S药物对映体的定量分离，R范围为4.36 - 5.64。这些肽和蛋白质硅胶整体固定相具有典型的HILIC分离行为和手性分离能力，这意味着它们不仅适用于未来的代谢研究，而且适用于药物对映体识别。
Glutathione (GSH)-, somatostatin acetate (ST)- and ovomucoid (OV)-functionalized silica-monolithic stationary phases were designed and synthesized for HILIC and chiral separation using capillary electrochromatography (CEC). GSH, ST and OV were covalently incorporated into the silica skeleton via the epoxy ring-opening reaction between their amino groups and the glycidyl moiety in γ-glycidoxypropyltrimethoxysilane (GPTMS) together with polycondensation and copolymerization of tetramethyloxysilane and GPTMS. Not only could the direction and electroosmotic flow magnitude on the prepared GSH-, ST- and OV-silica hybrid monolithic stationary phases be controlled by the pH of the mobile phase, but also a typical HILIC behavior was observed so that the nucleotides and HPLC peptide standard mixture could be baseline separated using an aqueous mobile phase without any acetonitrile during CEC. Moreover, the prepared monolithic columns had a chiral separation ability to separatedl-amino acids. The OV-silica hybrid monolithic column was most effective in chiral separation and could separatedl-glutamic acid (Glu) (the resolution R = 1.07),dl-tyrosine (Tyr) (1.57) anddl-histidine (His) (1.06). Importantly, the chiral separation ability of the GSH-silica hybrid monolithic column could be remarkably enhanced when using gold nanoparticles (AuNPs) to fabricate an AuNP-mediated GSH-AuNP-GSH-silica hybrid monolithic column. TheRofdl-Glu,dl-Tyr anddl-His reached 1.19, 1.60 and 2.03. This monolithic column was thus applied to separate drug enantiomers, and quantitative separation of all four R/S drug enantiomers were achieved with R ranging from 4.36 to 5.64. These peptide- and protein-silica monolithic stationary phases with typical HILIC separation behavior and chiral separation ability implied their promise for the analysis of not only the future metabolic studies, but also drug enantiomers recognition.