Preparation and characterization of hybrid-silica monolithic column with mixed-mode of hydrophilic and strong anion-exchange interactions for pressurized capillary electrochromatography

► A novel hybrid-silica monolith with HI-SAX mode was described. ► The monolith was optimized in details by a modified “one-pot” process. ► Column performances and HI-SAX mechanism were investigated. ► Monolith efficiently separated nucleotides, nucleic acid bases and nucleosides.