利用選擇性大量進樣陽離子搭配掃集式微胞電泳法針對人體血漿中安眠藥Zolpidem及其代謝物之同步分析研究

Simultaneous determination of zolpidem and its main metabolites in human plasma by cation-selective exhaustive injection and sweeping micellar electrokinetic chromatography

李明勳(Ming-Hsun Lee)

指導教授 ： 吳秀梅

高雄醫學大學/藥學院/藥學系碩士班/碩士(2016年)

摘要

本研究應用選擇性大量進樣陽離子搭配掃集式微胞電泳法
(cation-selective exhaustive injection-sweeping-micellar electrokinetic chromatography, CSEI-sweeping-MEKC) 對血漿中安眠藥zolpidem (Z) 及其主要代謝物zolpidem-6-carboxylic acid (Z6) 以及zolpidem phenyl-4-carboxylic acid (Z4) 進行同步分析。本法經簡單的檢品前處理過程後，於有效長度30 cm 之熔矽毛細管(內徑50 μm) 完成分離，首先通入分離用緩衝磷酸鹽溶液(40 mM, pH 2.5) 內含 20% ACN，接著通入一高導電度之緩衝磷酸鹽溶液(150 mM, 1 psi, 40 sec)，並以10 kV 取樣5 分鐘，再用-20 kV 電壓，通以掃集用緩衝磷酸鹽溶液(40 mM, pH 2.5) 內含 20% ACN 及100 mM SDS，將待測物堆積濃縮並分離。並於200 nm 波長下進行偵測。分析方法經確效後，Z 的定量範圍是5∼500 ng/mL，而Z6 及Z4 皆為25∼1000 ng/mL，其相關係數皆大於0.9996，顯示其具有良好的線性關係；同批間與異批間的精密度與準確度探討，相對標準偏差(RSD) 與相對誤差 (RE)皆小於 10.5%。本方法成功應用於臨床血漿檢品之檢測分析，並期盼未來能利用於偵測病人血中藥物濃度值之變化，以評估藥效及副作用。

關鍵字

選擇性大量進樣陽離子搭配掃集式微胞電泳法 ； 安眠藥 ； 使帝諾斯

並列摘要

In this study, a cation-selective exhaustive injection and sweeping micellar electrokinetic chromatography (CSEI-sweeping-MEKC) method was developed for simultaneous determination of zolpidem and its main metabolites (zolpidem-6-carboxylic acid, Z6; zolpidem phenyl-4-carboxylic acid, Z4) in human plasma. After sample pretreatment, phosphate buffer (40 mM, pH 2.5) containing 20% acetonitrile was filled into an uncoated fused silica capillary (30 cm, 50 μm I.D.), then high conductivity buffer (150 mM, 1 psi for 40 s) was followed. Electrokinetic injection (10 kV, 5 min) was used to load samples and to enhance the sensitivity. The stacking step and separation were performed at -20 kV and 200 nm by using phosphate buffer (40 mM, pH 2.5) containing 20% acetonitrile and 100 mM sodium dodecyl sulfate. During method validation, calibration plots were linear (r≧0.9996) over a range of 5-500 ng/mL for Z, and 25-1000 ng/mL for Z6 and Z4. The intra-batch and inter-batch assays demonstrated the good precision and accuracy due to the small RSD and RE values (≦10.5%). Futhermore, the developed method has been successfully applied to analyze real sample from patients, aiming to be helpful for adverse effect evaluation with drug concentration in plasma.

並列關鍵字

CSEI-sweeping-MEKC ； hypnotic drugs ； Stilnox

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