利用毛細管電泳法進行血清中carbamazepine及其五種代謝物的濃度分析

Analysis of carbamazepine and its five metabolites in serum by capillary electrophoresis

林妍瑜(Yan-Yu Lin)

指導教授 ： 吳秀梅

高雄醫學大學/藥學院/藥學研究所/碩士(2012年)

摘要

本實驗利用緩衝溶液導電度的差異結合大體積樣品進樣與掃集技術進行樣品堆積，對血清檢體中的carbamazepine (CBZ)及其五種代謝物10,11-dihydro-10,11-epoxycarbamazepine、10,11-dihydro-10,11-dihydroxycarbamazepine、2-hydroxycarbamazepine、3-hydroxycarbamazepine與10,11-dihydro-10-hydroxycarbamazepine同時分析。將待測物溶於低濃度磷酸緩衝液，以壓力大體積方式進樣(10 psi, 20 s)，打入充滿高濃度背景磷酸緩衝液的毛細管中(50/60 cm)。在施加- 20 kV分析電壓後，於緩衝液導電度的差異下導致待測物在緩衝液界面移動率減緩，而大體積的樣品則藉由電滲透流的回推力進行濃縮，同時以添加有50 mM SDS與15%甲醇的掃集磷酸緩衝液往偵測端進行掃集而完成樣品堆積作用，搭配紫外光偵測器以214 nm進行偵測。分析方法經確效後，CBZ定量範圍為0.03 - 25 μg/mL，五種代謝物皆為0.03 - 3 μg/mL，其相關係數皆大於0.997。此方法中六種分析物的偵測極限(S/N=3)皆為0.01 μg/mL，相較於MEKC的偵測感度可以提升約300倍，且此分析方法最後實際應用於六位每日投予200 mg CBZ之患者其血中濃度監測，其濃度範圍於2.88到5.82 μg/mL。

關鍵字

毛細管電泳 ； Carbamazepine ； Carbamazepine代謝物 ； 血清 ； 大體積樣品進樣掃集堆積

並列摘要

To improve the sensitivity, this study used the different conductivity buffers and LVSS-sweeping for carbamazepine (CBZ) and its five metabolites analyzing. In this research, the capillary (50/60 cm) was filled with high concentration of background electrolyte added 15% methanol, then a large-volume of sample, reconstituted in low concentration of sample matrix added 5% methanol, was injected into the capillary. When high voltage was applied (- 20 kV), the analytes decelerated at the boundary between low and high-conductivity buffer; meanwhile the large-volume of sample was stacked by electroosmotic flow pushing it backward and formed a narrow sample zone. At the same time, the sweeping procedure was carried out by sweeping buffer, added 15% methanol and 50 mM SDS, and the detection was performed by UV detector with 214 nm. The method was validated for linearity (r≧0.997), precision and accuracy. The calibration curves were established for CBZ and its metabolites between 0.03 - 25 μg/mL and 0.03 - 3 μg/mL, respectively. The limits of detection (S/N=3) were 0.01 μg/mL for each analyte. Compared with MEKC, this system can improve the sensitivity about 300-fold. Finally, this method was successful applied to 6 patients, who have taken 200 mg CBZ daily, and drug levels were found from 2.88 to 5.82 μg/mL.

並列關鍵字

Capillary electrophoresis ； Carbamazepine ； Metabolites of carbamazepine ； Serum ； Large volume sample stacking-sweeping

參考文獻

參考文獻

[1] N. S. Baath, J. Hong, and S. P. Sattar. Possible carbamazepine toxicity with terbinafine (2006) Can J Clin Pharmacol 13:e228-e231.

[2] A. Nakamura, K. Mihara, G. Nagai, T. Suzuki, and T. Kondo. Pharmacokinetic and pharmacodynamic interactions between carbamazepine and aripiprazole in patients with schizophrenia (2009) Ther Drug Monit 31:575-578.

[3] K. Octani, M. Ishida, N. Yasui, T. Kondo, K. Mihara, A. Suzuki, H. Furukori, S. Kaneko, and Y. Inoue. Interaction between carbamazepine and bromperidol (1997) Eur J Clin Pharmacol 52:219-222.

[4] A. C. Leon, D. A. Solomon, C. Li, J. G. Fiedorowicz, W. H. Coryell, J. Endicott, and M. B. Keller. Antiepileptic drugs for bipolar disorder and the risk of suicidal behavior: a 30-year observational study (2011) Am J Psychiatry 168:1-7.