The development of monodisperse hybrid silica microspheres with highly regular pore structure and uniform distribution of functional groups have significant value in the biomolecular separation field. In this work, the short range ordered pore channels are precisely constructed onto the non-porous silica microsphere surface by a bi-phase assembly method, and the cylindrical silica channel introduced a plethora of vinyl groups by "one-pot" co-condensation to form vinyl hybrid silica shell. As hydrophilic interaction chromatography (HILIC) stationary phase, the vinyl hybrid core-shell silica microsphere is simply modified with zwitterion glutathione (SiO2@SiO2-GSH), in which the HILIC enrichment process is significantly shortened due to its specific porous characteristics. Most importantly, SiO2@SiO2-GSH microsphere can enrich 2186 N-glycopeptides from the rat liver protein digest within 2 min, which mapped to 806 glycoproteins. Compared with HILIC enrichment result within 1 h, the glycoproteins and glycopeptides overlap are 88.3% and 79.1%, performing excellent reproducibility. The short range ordered channels onto the silica microsphere surface exhibit excellent mass transfer efficiency, so the developed bi-phase assembly method is expected to design more advanced hybrid silica materials for other urgently fields.
Keywords: bi‐phase; chromatographic separation; core–shell; hybrid silica; self‐assembly.
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