Preparation of a Hybrid Cation-Exchange Monolith and Its Application for the Separation of Proteins by High Performance Liquid Chromatography

Abstract

A hybrid cation-exchange monolith for high performance liquid chromatography (HPLC) was prepared by a one-pot synthesis with reverse-atom transfer radical polymerization, in which sodium bisulfite was used as the inorganic material, methyl methacrylate was used as organic material, 2, 2'-Azobisisobutyronitrile as initiator, and ferric trichloride as the inorganic catalyst. Moreover, sodium bisulfite was used to provide the sulfonic group. The conditions of polymerization were optimized. The chemical groups of the monolith were assayed by Fourier transform infrared spectroscopy; the morphology of monolithic material was investigated by scanning electron microscopy; the pore size distribution was determined by a mercury porosimeter; and the mechanical stability and permeability of the monolith were investigated by the back-pressure drop at several linear velocities. Finally, the monolith was used to separate human serum albumin from human plasma in conjunction with HPLC in 5 min, and the influences of buffer concentration and pH value on the elution of human serum albumin were investigated. In addition, the monolith was applied to separate lysozyme from egg whites in a short time (4 min) and separate a mixture of aromatic compounds.