電場放大電動取樣及短端注射毛細管電泳法對人體血漿中meperidine及其代謝物normeperidine之分析研究

Simultaneous determination of meperidine and normeperidine in plasma by field amplified sample injection and short-end capillary electrophoresis

許育栽(Yu-Tsai Hsu)

指導教授 ： 吳秀梅

高雄醫學大學/藥學院/藥學研究所碩士在職專班/碩士(2014年)

摘要

本研究開發毛細管電泳分析法，利用線上堆積技術”電場放大電動取樣”及短端注射方式來同時分析人體血漿中meperidine (MP)及其代謝物normeperidine (NMP)。MP會結合到中樞神經系統的鴉片受體而產生鎮痛效果，但長期連續投與MP將因其代謝物NMP的蓄積，而造成中樞神經毒性，因此有必要建立有效之方法來監測其體內濃度。本法在毛細管內預先注入高濃度的背景緩衝溶液使其充滿整個毛細管後，再注入一段去離子水，接著以電動取樣大量注入樣品，在施加−20 kV分析電壓後，於緩衝液導電度的差異下減緩待測物在緩衝液界面移動率而完成樣品堆積作用，搭配紫外光偵測器以214 nm進行偵測。本法相較於壓力取樣(2 psi, 10 s)可提升感度約350倍；而短端注射毛細管電泳法可以有效縮短分析時間至2分鐘內。此法經確效後，偵測極限定義為3倍訊號/雜訊值(S/N=3)，MP與NMP皆為3 ng/mL，而定量範圍為10-500 ng/mL，其相關係數(r)均大於0.99。對於血漿中的MP與NMP，進行同日樣品內(N=3)與異日樣品間(N=5)分析方法之準確度與精密度皆小於8%。此方法目前已成功地應用於實際MP肌肉注射50 mg之人體檢體之血中濃度測定，MP於30分鐘達到最高血漿濃度，NMP則於60分鐘，而血漿濃度MP為180.7 ± 9.8 ng/mL，NMP為64.9 ± 3.3 ng/mL，經計算後得MP之半衰期分別為3.3，NMP為18.5小時，皆與藥物動力學參數相符合，未來可做為臨床分析MP與NMP血中濃度之應用。

關鍵字

配西汀 ； 去甲基配西汀 ； 短端注射毛細管電泳法 ； 電場放大電動取樣 ； 血漿

並列摘要

A stacking capillary electrophoresis method, field amplified sample injection and short-end injection procedure (FASI-SEIP), was developed for the determination of meperidine (MP) and normeperidine (NMP) in human plasma. MP produces analgesia by binding opioid receptors in the central nervous system (CNS). But continuous administration may result in the accumulation of NMP, responsible for CNS toxicities. It is important to establish one assay method. In this study, the capillary was filled with background electrolyte, then a water plug was injected. The electrokinetic injection was carried out, followed by a high voltage of −20 kV for separation. When analytes passed the high field region (water plug) and enter the low field region (background electrolyte), they accumulated in the interface. Finally, the analytes was stacked and separated within 2 min. The LOD (S/N= 3) was 3 ng/mL for MP and NMP, respectively. The sensitivity was about 350 folds comparing with hydrodynamic injection mode (2 psi, 10 s). During method validation, calibration curves were linear (r≧0.99) over a range from 10-500 ng/ml for MP and NMP. As the precision and accuracy assays, the absolute values of relative standard deviation (RSD) and relative error (RE) in intraday (n=3) and interday (n=5) observations were less than 8%. This method was successfully applied to real samples. After 50 mg intramuscular administration, the maximum concentration of MP was 180.7 ± 9.8 ng/mL at 30 min, and that of NMP was 64.9 ± 3.3 ng/mL at 60 min. The half-lives are 3.3 hours for MP and 18.5 hours for NMP. It was coincident to its pharmacokinetic profile in human. This method was feasible for clinical detection of MP and NMP
in human plasma.

並列關鍵字

meperidine ； normeperidine ； short-end capillary electrophoresis ； field amplified sample injection ； human plasma

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