Zhao Licong, Yang Limin, Wang Qiuquan
Department of Chemistry & the Key Laboratory of Spectrochemical Analysis and Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, PR China.
Department of Chemistry & the Key Laboratory of Spectrochemical Analysis and Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, PR China.
J Chromatogr A. 2016 May 13;1446:125-33. doi: 10.1016/j.chroma.2016.04.014. Epub 2016 Apr 7.
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 separate dl-amino acids. The OV-silica hybrid monolithic column was most effective in chiral separation and could separate dl-glutamic acid (Glu) (the resolution R=1.07), dl-tyrosine (Tyr) (1.57) and dl-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. The R of dl-Glu, dl-Tyr and dl-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.
设计并合成了谷胱甘肽（GSH）、醋酸生长抑素（ST）和卵类黏蛋白（OV）功能化的硅胶整体固定相，用于采用毛细管电色谱（CEC）的亲水相互作用色谱（HILIC）和手性分离。通过GSH、ST和OV的氨基与γ-缩水甘油氧基丙基三甲氧基硅烷（GPTMS）中的缩水甘油基部分之间的环氧开环反应，以及四甲氧基硅烷和GPTMS的缩聚和共聚反应，将GSH、ST和OV共价结合到硅胶骨架中。制备的GSH-、ST-和OV-硅胶杂化整体固定相上的电渗流方向和大小不仅可以通过流动相的pH值进行控制，而且还观察到了典型的HILIC行为，从而在CEC过程中使用无任何乙腈的水性流动相时，核苷酸和HPLC肽标准混合物可以实现基线分离。此外，制备的整体柱具有分离dl-氨基酸的手性分离能力。OV-硅胶杂化整体柱在手性分离方面最有效，能够分离dl-谷氨酸（Glu）（分离度R = 1.07）、dl-酪氨酸（Tyr）（1.57）和dl-组氨酸（His）（1.06）。重要的是，当使用金纳米颗粒（AuNP）制备AuNP介导的GSH-AuNP-GSH-硅胶杂化整体柱时，GSH-硅胶杂化整体柱的手性分离能力可以显著提高。dl-Glu、dl-Tyr和dl-His的R分别达到1.19、1.60和2.03。因此，该整体柱被应用于分离药物对映体，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至5.64。这些具有典型HILIC分离行为和手性分离能力的肽和蛋白质-硅胶整体固定相不仅表明它们在未来代谢研究分析中具有前景，而且在药物对映体识别方面也具有前景。
