Silica monolith particles with sizes of 2-5 μm and pore sizes of 20-60 nm were obtained by grinding, flotation, pseudomorphic transformation, and hydrothermal treatment of the silica monolith prepared by the sol-gel method. The pseudomorphic transformation was performed with a dual micellar templating system consisting of Capstone FS-66, a partially fluorinated anion surfactant, and cetyltrimethylammonium bromide (CTAB), a commonly used cation surfactant. Hydrothermal treatment with a sodium carbonate solution was adopted to further expand the pore size. Scanning electron microscopy (SEM) images and N2 adsorption-desorption isotherm measurement results of the silica monolith particles before and after the treatments clearly demonstrated the changes in morphology caused by these treatments. Afterward, a long-chain polyethylene glycol (PEG) containing silane was bonded on the surface of the as-prepared particles, and the resulting products were characterized by elemental analysis and FT-IR spectroscopy analysis, and evaluated by high performance liquid chromatography (HPLC). Elemental analysis and thermogravimetric analysis (TGA) of the bonded stationary phase revealed that the bonding amount of PEG on the silica surface is about 8%. It has been shown that silica monolith particles can be treated and modified for the separation of proteins in size exclusion chromatography mode. It is also demonstrated that the bonded stationary phase can be used for the separation of ribonuclease A and lysozyme in hydrophobic interaction chromatography mode, and for the separation of highly polar compounds (picolinic acid, levodopa, melamine, and catechol) in hydrophilic interaction chromatography mode. The results indicate the versatility of the PEG-bonded stationary phase and its promising application to multi-modal separation in HPLC.
Keywords: high performance liquid chromatography (HPLC); multi-modal stationary phase; protein separation; pseudomorphic transformation; silica monolith particles.