Preparation Of Poly(high Internal Phase Emulsion)monolith And Its Application To The Separation Of Proteins

High performance liquid chromatography (HPLC) plays an important role in pharmaceutical analysis at present. Column is the heart of the HPLC, so the key step to improve the separation technique is the development of chromatographic media. Because of the advantages of simple preparation and good permeability, the monolithic column has been applied widely in HPLC analysis. Therefore, this thesis focuses on the fabrication and characterization of novel monolithic column for chromatography analysis.In this paper, a summary concerned with the development and application of monolith was introduced firstly. Using vinyl ester resin as monomer, ethylene glycol dimethacrylate (EGDMA) as the crosslinker, potassium persulfate as initiators and PF127 as emulsifier, a poly(vinyl ester) resin monolith was prepared by"in-situ"process. The main factors affecting the porous structure, the mechanical stability and the penetrability of the monolith were investigated. Moreover, the monolithic column was modified into strong cation-exchange and Lysozyme was separated from chicken egg white with high resolution in a short time, which demonstrated the potential of poly(vinyl ester) resin monolith for the fast separation of proteins.Using glycidyl methacrylate as monomer, crosslinker and other reagents as above, a novel porous polymeric monolith based on poly(high internal phase emulsion) was prepared. The main factors affecting the porous structure and the mechanical stability of monolithic columns were investigated. These unique properties, together with high porosity, good mechanical property and chemical modification of the surface make themselves superior in monolithic medium applications. Additionally, the separation capabilities of this monolith in conjunction with HPLC were investigated. Immunoglobulin could be separated from human plasma and chicken egg yolk with high resolution on the hydrophobic interaction chromatographic column within 2 min. Moreover, fast separation of a two mode protein mixture (IL-18 and lysozyme) on the monolith was achieved within 1.5 min at a velocity of 1445 cm/h. As a result, good separation was achieved, and stable low back pressure drop was ensured at high throughput elution with an even longer column.