利用電場加大樣品堆積 ( Field-amplified sample stacking ) 建立簡易毛細管電泳分析法對doxorubicin及其主要活性代謝物doxorubicinol血中濃度同時分析之研究

Simultaneous determination of doxorubicin and active metabolite doxorubicinol in plasma by capillary electrophoresis and field-amplified sample stacking

顏妙如(Miao-Ju Yen)

指導教授 ： 陳素惠

高雄醫學大學/藥學院/藥學研究所/碩士(2009年)

摘要

本論文乃利用電場加大樣品堆積之毛細管電泳法( Field-amplified sample stacking ; FASS )，搭配UV作為偵測器(CE-UV)，針對人體血漿中抗癌藥物doxorubicin及其代謝物doxorubicinol，建立一個簡單、同時並具有選擇性之定量分析方法。此方法分析條件為: 血漿檢品經過前處理之後，以metformin做為內部標準品進行分析，緩衝溶液組成包括有50 mM磷酸鹽緩衝溶液( pH 6.5 ) 內含16% ethylene glycol及60% methanol。待測物 ( doxorubicin及其代謝物doxorubicinol ) 檢量線範圍為20.0 - 100.0 ng/mL，而二者之檢出極限分別為5 ng/mL (doxorubicin) 及10 ng/mL (doxorubicinol) ( s/n=3，電壓取樣10 kV 30秒 )。影響分離的因素包括: 磷酸鹽緩衝溶液的pH值和濃度，ethylene glycol的濃度、有機溶媒添加之體積量加以探討。此分析方法可應用於血漿中doxorubicin及doxorubicinol之含量分析。

關鍵字

毛細管電泳 ； 電場加大樣品堆積 ； 抗癌藥

並列摘要

A field-amplified sample stacking and capillary electrophoresis with UV detection (CE-UV) is described for the determination of doxorubucin and doxorubicinol in human plasma. After sample pretreatment, field-amplified sample stacking method is applied for sensitive improvement. The sample was employed with an electrokinetic injection of 10 kV for 30 sec. CE separation of doxorubicin and doxorubicinol from human plasma was performed at 250C using a background electrolyte consisting of phosphate buffer (50 mM, pH 6.5) containing 16% ethylene glycol and methanol 60%. Using metformin as an internal standard (I.S.), the linear range of the method for the determination of doxorubicin and doxorubicinol was over 20.0-100.0 ng/mL. The LOD of the doxorubibin and doxorubicinol in human plasma was 5 ng/mL(S/N=3, 10 kV, 30 sec) and 10 ng/mL (S/N=3, 10 kV, 30 sec) respectively. For optimization of the procedure, the pH and strength of phosphate buffer, the volume of ethylene glycol and methanol would be discussed.
Application of the method to the determination of doxorubicin and doxorubicinol in human plasma proved to be feasible.

並列關鍵字

Capillary electrophoresis ； field-amplified sample stacking ； antineoplastic agent

參考文獻

1.G.N Hatobagyi. Anthracyclines in the treatment of cancer. An overview, Drugs, 1997,54(suppl.4), 1-7.

2.R.M. O’Bryan, L.H. Baker, J.B. Gottlieb, S.E. Rivkin, S.P. Balcerzak, G.N. Grumet, S.E. Salmon, T.E. Moon, B. Hoogstraaten, Dose response evaluation of adriamycin in human neoplasia,Cancer,1977,39, 1940-1948.

3.R. Danesi, S. Fogli, A. Gennari, P. Conte, M. Del Tacca, Pharmacokinetic-pharmacodynamic relationships of the anthracycline anticancer drugs, Clin. Pharmacokinet., 2002 , 41 (6) , 431-/444.

4.DJ. Stewart, D. Grewaal, RM. Green, N. Mikhael, R. Goel, VA. Montpetit, MD. Redmond, Concentrations of doxorubicin and its metabolites in human autopsy heart and other tissues, Anticancer Research, 1993, 13(6A), 1945-1952.

5.S. Fogli, D. Danesi, F. Innocenti, A. Di Paolo, G. Bocci, C. Barbara, M. Del Tacca, An improved HPLC method for therapeutic drug monitoring of daunorubicin, idarubicin, doxorubicin, epirubicin, and their 13-dihydro metabolites in human plasma, Ther. Drug Monit., 1997, 21(3), 367.