毛細管電泳法對阿茲海默症及帕金森氏症治療藥物與胰島素製劑之分析研究與應用

本論文的主題是針對藥物發展出簡單及高感度的分析方式，主要研究包括利用微胞電動層析法針對低血中濃度阿茲海默症及帕金森氏症藥物達到微量分析的目的，及利用毛細管區帶電泳法分離insulin與相差一個胺基酸的insulin aspart。主要成果摘錄如下: I. 毛細管電泳法對阿茲海默症及帕金森氏症治療藥物之微量分析研究與應用研究內容包括以線上濃縮的方式對乙烯膽鹼酶抑制劑(acetylcholine-esterase inhibitor, AChEI)-donepezil達到有效的微量分析；以及配合化學衍生的方式分析人體血漿中N-methyl D-aspartate (NMDA)受體抑制劑memantine及其結構相似用於治療帕金森氏症之amantadine的濃度，以應用於臨床上醫師對於病患治療劑量的參考。 1. 毛細管電泳法對乙烯膽鹼酶抑制劑(AChEI)-donepezil之微量分析研究本研究以liquid-liquid extraction (LLE)為血漿樣品前處理方式，將donepezil由血漿萃取至有機層，吹乾後回溶。再以毛細管電泳配合電場加大樣品濃縮法(filed amplified sample stacking，FASS)進行線上濃縮及分析。此方法應用於阿茲海默症病人血漿中donepezil濃度監測及健康受試者服藥後的藥物動力學參數計算。 2. 毛細管電泳法配合化學衍生對人體血漿中memantine及amantadine之同時分析本研究利用化學衍生的方式配合毛細管電泳法同時分析memantine及amantadine。血漿樣品以LLE前處理後吹乾回溶於borate緩衝液，加入衍生試劑CFSE (6-carboxyfluorescein N-hydroxy-succinimide ester)進行衍生反應。衍生產物以MEKC模式使用中性界面活性劑Brij-35®達到有效分離。此方法應用於阿茲海默症病人血漿中memantine及帕金森氏症病人amantadine的濃度監測。 II. 毛細管電泳法對regular insulin及insulin aspart的同時分析及其在分析市售製劑上之應用此部分研究內容為建立一簡單的毛細管區帶電泳法(capillary zone electrophoresis)進行insulin與insulin aspart的同時分析。Regular insulin與insulin aspart在結構上非常接近，只差在B28位置上的胺基酸，將proline以aspartic acid取代即為insulin aspart。本研究分析條件為: 80 mM phopshate緩衝溶液(pH 6.5)，實驗上探討影響分離的參數，包括緩衝液的濃度和pH值。並將建立之分析方法應用於製劑分析。

關鍵字

毛細管電泳；阿茲海默症；帕金森氏症

並列摘要

The theme of this study is to develop a simple, rapid and high sensitive analytical method for the determination of therapeutic agents. The main works included using MEKC method for the trace analysis of anti-Alzheimer’s disease and anti-Parkinson’s disease agents with low plasma concentrations; or using CZE method for the simultaneous determination of insulin and insulin aspart, which were different in single amino acid. The results are summarized as follow: I. Trace Analysis of Anti-Alzheimer’s Disease and Anti-Parkinson’s Disease agents by capillary electrophoresis The works of this study included the development of a CE method with on-line sample stacking to achieve trace analysis of acetylcholine esterase inhibitor (AChEI) agent – donepezil in plasma; and using chemical derivatization combined with CE method to analysis N-methyl D-aspartate (NMDA) receptor antagonist – memantine and its structurally similar compound – amantadine in human plasma. The developed methods were applied in clinical as the reference for physicians to evaluate the therapeutic dosage of patients. 1. Trace analysis of acetylcholine esterase inhibitor (AChEI) – donepezil by capillary electrophoresis In this work, liquid-liquid extraction (LLE) was used as the pretreatment method to extract donepezil from plasma to organic layer, which was dried and the residue was reconstructed. CE coupled with field amplified sample stacking (FASS) was used to perform on-sample stacking and analysis. The method was applied for monitoring plasma concentration of donepezil in Alzheimer’s disease patients; and evaluates the pharmacokinetic data of a health volunteer. 2. Simultaneous determination of memantine and amantadine in human plasma by capillary electrophoresis coupled with chemical derivatization A CE method combined with derivatization was developed for the simultaneous determination of memantine and amantadine. The plasma sample was pretreated with LLE, the organic layer was dried and reconstructed with borate buffer to perform chemical derivatization with the labeling reagent – CFSE. The derivatized products were efficiently separated by a MEKC method with non-ionic surfactant Brij-35®. The developed method was applied for the monitoring plasma concentration of memantine and amantadine in Alzheimer’s disease and Parkinson’s disease patients. II. Simultaneous determination of insulin and insulin aspart by capillary electrophoresis The main work of this study is to develop a simple capillary zone electrophoresis for the simultaneous determination of insulin and insulin aspart. The structures of regular insulin and insulin aspart are very similar, the amino acid on B28 position was replaced from proline to aspartic acid to form insulin aspart. The BGE used in the study was 80 mM phosphate buffer (pH 6.5), and different parameters were discussed including the concentration and pH value of buffer to optimize the separation condition. The developed method was applied for the analysis of pharmaceutical formulation.