Zwitterionic silica-based monolithic capillary columns for isocratic and gradient hydrophilic interaction liquid chromatography

► We prepare zwitterionic monolithic capillary columns intended for HILIC separations. ► Columns were evaluated under the isocratic and gradient separation conditions. ► Prepared columns showed high permeability and efficiency. ► Successful separation of modified nucleobases and nucleosides was achieved. This study introduces zwitterionic monolithic capillary columns intended for isocratic and gradient HILIC separations. Silica-based monolithic capillary columns (100μm×150mm) prepared by acidic hydrolysis of tetramethoxysilane in the presence of polyethylene glycol and urea were modified by a sulfoalkylbetaine zwitterion ([2-(methacryloyloxy)ethyl]-dimethyl-(3-sulfopropyl)-ammonium hydroxide) to HILIC stationary phase. The prepared columns were evaluated under the isocratic and gradient separation conditions using a standard mixture containing nucleic acid bases, nucleosides, and 2-deoxynucleosides. Mobile phases contained high concentration of acetonitrile (95–85%, v/v) and 5–50mM ammonium acetate or ammonium formate in the pH range of 3–6. The synthesized columns showed a long-term stability under the separation conditions while the high permeability and efficiency originating from dual structure of the silica monolith were preserved. The relative standard deviations (RSDs) for the retention times of tested solutes were lower than 2% under the isocratic conditions and lower than 3.5% under the gradient conditions.