Preparation Of Polymer Monoliths By Two Kinds Of "Living"/Controlled Radical Polymerization And Their Applications In The Separation Of Proteins

As an important separation and analytical method, high performance liquid chromatography (HPLC) plays an important role in life science, medical science, chemical industry and environmental science. It is of great significance to research, develop and improve chromatographic media, because column is the heart of the HPLC. A lot of advantages, high permeability, porous structure and large specific surface area, make monolithic column have a rapid development, especially in the separation and analysis biomacromolecules.At first, the classification, the characteristics and preparation of monolithic column were introduced, and classification and their mechanisms were explained.Secondly, monolithic column was prepared with vinyl ester resin as monomer and crosslinker by RITP/RATRP, and its construction, permeability and mechanical stability were investigated. Lysozyme was separated from chicken egg white with high resolution in a short time, in addition, the influences of buffer concentration and pH value on elution were investigated, respectively.Finally, monolithic column was prepared with methyl methacrylate (MMA) as monomer, ethylene glycol dimethacrylate (EDMA) as the crosslinker, carbon tetrachloride (CCl4) as initiators, ferrous chloride (FeCl2) as catalyst natrium bisulfurosum (NaHSO3) as inorganic adjunct, by Atom Transfer Radical Polymerization (ATRP). The main factors affecting the preparation of monolithic columns were investigated. Besides, the mixture of Lys, papain and bovine serumal bumin (BSA) were separated by the monolith with good resolution and reproducibility. In addition, the influences of buffer concentration and pH value on elution were investigated, and the separation mechanism was studied.