Preparation and chromatographic evaluation of the hydrophilic interaction chromatography stationary phase based on nucleosides or nucleotides

In this work, a series of novel hydrophilic interaction chromatography (HILIC) stationary phases were prepared by grafting nucleosides or nucleotides on the surface of silica gel. Firstly, the silica was modified with 3-glycidoxypropyltrimethoxysilane (GPTMS). And then nucleosides or nucleotides were bonded on the surface of GPTMS-modified silica through the epoxy-amine ring-opening reaction to provide four HILIC materials. These obtained stationary phases were successfully characterized by Fourier transform-infrared spectroscopy (FT-IR) and elemental analysis (EA), respectively. Effects of column temperature, water content of the mobile phase, pH and buffer concentration on the retention behavior of these HILIC materials and the corresponding separation mechanism were evaluated using various nucleosides and nucleobases, respectively. In addition, polar and hydrophilic compounds such as amino acids and water-soluble vitamins were successfully separated using the corresponding columns, showing application potential for the separation of bioactive substances.