Preparation of Strong Cation Exchange Packings Based on Monodisperse Hydrophilic Non-porous Resins and Their Application for Fast Separation of Proteins

Monodisperse, 3.0 μm non-porous hydrophilic poly (glycidylmethacrylate-co-ethylenedimethacrylate) particles were prepared by one-step swelling and polymerization method. The particles were modified to be a strong cation exchange (SCX) stationary phase for high performance liquid chromatography (HPLC) in following steps. First, the particles were completely hydrolyzed. Second, the hydrolyzed particles were reacted with epichlorhydrin followed by another hydrolysis of the newly introduced epoxide groups. Third, the particles were reacted with chlorosulfonic acid. The SCX stationary phase was evaluated in light of the ion exchange property, separability and hydrophilicity on the separation and retention of proteins in detail. Four proteins were quickly separated in 1.0 min linear gradient elution using the synthesized SCX stationary phase. It was found that it followed ion exchange chromatographic (IEC) retention mechanism. The SCX resins were used for the fast purification of lysozyme from egg white and cytochrome-C from pig heart in 3.0 min with only one step. The results obtained were satisfactory.