本研究的目的是在管柱層析中設計一個固定相作為膽固醇之分析。分子模版高分子提供與模版分子互補的鍵結位置，而此高辨識能力可作為管柱層析之固定相。懸浮聚合所製備之分子模版高分子具較適於管柱層析的多孔性球狀結構。聚合時的攪拌速率能輕易地控制顆粒之粒徑。在聚合過程中，連續相（水）會影響官能基單體與模版之間氫鍵鍵結。為了防止水的影響，在聚合過程中膽固醇甲基丙烯酸酯的加入，產生共價模印的模版─單體錯合物。此外，使用鄰苯二甲酸二(2-乙己基)酯 ：正癸烷 = 77：23體積比作為造孔劑的分子模版高分子EMPD410H呈現破碎狀，表面具有多孔性、高表面積（277.4 m2/g）特點，對膽固醇吸附量（3.993 μmole/g）比使用正丁所製備之分子模版高分子EMBD410H高（2.443 μmole/g）。使用正丁作為造孔劑，分子模版高分子在聚合後可形成球狀，但是表面積只有0.2 m2/g。
以直徑為4-12 μm的分子模版高分子填充管柱可以有效分離膽固醇與雌素二醇。膽固醇結構為疏水性，移動相中乙比水的的比例從19:0提高到19:2時，膽固醇的滯留時間從21.2 min增加為27.7 min，分離因子從1.26增加為4.43，解析度從無法計算變為1.20，理論板數從69降低至37。此外，移動相中水的比例低時，膽固醇在無模印的分子模版高分子作為固定相時，會先流出管柱而雌素二醇則較慢流出，此結果與分子模版高分子相反。另外，管柱EMPD1190H相較於所有的管柱有較好的分離因子（16.02）與解析度（3.44），伴隨著高表面積與良好的填充。
相較於懸浮聚合，塊狀聚合所製備之顆粒在管柱層析上有些許問題，例如波峰變寬、波峰拖曳以及高操作壓力（乙比水的比例19:2，流速0.5 mL/min下，管柱壓力65 kg/cm2）。管柱分離之研究也顯示管柱溫度20℃提高至60℃時，理論版數從37增加至217。

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.

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