Application of cholesterol-imprinted polymer prepared by suspension polymerization on chromatography

• Main Authors: Chih-Wei Kuo, 郭致瑋
• Other Authors: Ming-Chang Yang
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/76222991679015239462

碩士 === 國立成功大學 === 化學工程學系碩博士班 === 96 === The purpose of this study was to design a stationary phase in chromatography to analyze cholesterol. Molecular imprinted polymers provides the complementary binding site(s) for the template molecule and could be used as a stationary phase in chromatography due to its good specific recognition ability. Molecular imprinted polymers prepared by suspension polymerization could form the spherical macroporous particles, which might avoid the peak broadening and tailing in chromatography. The particle size could be easily controlled by the rate of agitation during the polymerization. The hydrogen bond between functional monomer and template might be affected by the continuous phase, water, during the polymerization, and can be avoided by the application of cholesteryl methacrylate in the polymerization to form the covalent imprinting with the template-monomer complex. In addition, using dioctyl phthalate:n-decane = 77:23 (v/v) as porogenic solvent to synthesize imprinted polymer EMPD410H, the polymer would become monolith and porous with surface areas up to 277.4 m2/g. The uptake of cholesterol (3.993 μmole/g) with EMPD410H was higher than that with the imprinted polymer EMBD410H prepared by porogenic solvent, butyronitrile (2.443 μmole/g). Using butyronitrile as porogenic solvent, the polymer would become spherical particle with surface areas as less as 0.2 m2/g. The column packed with 4~12μm-imprinted polymers separated cholesterol from β-estradiol effectively. The structure of cholesterol was hydrophobic. When the mobile phase, acetonitrile to water ratio changed from 19:0 to 19:2, the retention time of cholesterol was increased from 21.2 min to 27.7 min, the separation factor was increased from 1.26 to 4.43, and the plate number was decreased from 69 to 37 with the resolution of 1.20. For the non-imprinted polymers as stationary phase, cholesterol would elute first and β-estradiol second in the low water content. The result was different from that with imprinted polymers. Furthermore, the column with EMPD1190H polymer, with higher surface area and better packing, gave higher separation factor (16.02) and resolution (3.44) than other columns. Compared to the polymer prepared with the suspension polymerization, the particles prepared by bulk polymerization had several problems in chromatography, such as broaden and tailed peak with high performance pressure (at acetonitrile : water = 19:2, flow rate, 0.5 mL/min, column pressure, 65 kg/cm2 ). The study on the chromatographic separation also revealed that the increase in column temperature from 20℃ to 60℃ would increase the plate number from 37 to 217, and that the decrease in flow rate increased the resolution.