Preparation Of A Novel Hybrid Monolithic Column And Application In The Separation Of Proteins

At present, in the field of pharmaceutical analysis, High performance liquid chromatography (HPLC) plays an important role and column is the heart of the HPLC. So, development of new chromatographic media has important practical significance. There are many advantages of polymer monolith. The structure and pore-size distribution of the fixed phase can be adjusts expediently by altering the ratio of the porogen, monomer and cross linking agent. And the polymer monolithic column has the certain superiority in the separation biological macromolecule aspect. Therefore, this paper focuses on the fabrication and characterization of novel monolithic column for chromatography.At first, the classification, the preparation and application of polymeric monolithic column were introduced. Moreover, the advantages of hybrid materials the was described.In this work, based on ATRP, poly ( glycidyl methacrylate-co-ethyleneglycol dimethacrylate)hybrid monolith, which catalyzed by FeCl2/CCl4, was prepared. The mixture system of Glycidyl methacrylate as monomer, Ethylene dimethacrylate as cross linking agent, dodecyl alcohol and methyl alcohol as porogen, carbon tetrachloride as initiator and FeCl2 as catalyst was made to the hybrid monolith. The monolithic column was characterized by various technique. The main factors affecting the porous structure, the mechanical stability and the penetrability of the monolith were investigated. The results show that the column has good mechanical tolerance. Lysozyme standard was successful separated by the column, and the condition of separation was optimized.Finally, the separation of albumin egg have been explored, and the condition was optimized. And then, the actual samples, albumin egg in egg, was isolated by the column. As a result, good separation was achieved.