毛細管電泳法對抗痛風及降尿酸劑之分析研究

本論文之主題在於建立毛細管電泳法 ( capillary electrophoresis , CE )對多種抗痛風及降尿酸藥物之同時定量分析。首先探討區帶毛細管電泳法 (capillary zone electrophoresis) 對allopurinol、benzbromarone、orotic acid、oxypurinol、probenecid、sulfinpyrazone及colchicine 七種藥物之同時分析。初步結果顯示，在硼酸鹽緩衝溶液 ( 50 mM ; pH 9.00 )，使用15 kV的分離電壓，偵測波長為214 nm條件下，除了colchicine外，能分離其餘六種待測物，且具定量性，其定量範圍均在20 - 100 μM。進而利用微胞電動毛細管層析法( micellar electrokinetic chromatography, MEKC ) 分別針對上述七種待測物進行同時分析研究。結果顯示以硼酸鹽緩衝溶液 ( 45 mM; pH 9.00 )、20 mM sodium dodecyl sulfate ( SDS )，使用20 kV的分離電壓，偵測波長為214 nm，能分離上述等七種待測物。其中allopurinol、oxypurinol、orotic acid及probenecid定量範圍在10–200 μM，benzbromarone、colchicine、sulfinpyrazone定量範圍則為5-200 μM。偵測極限 (limit of detection) : allopurinol 2.0 μM; benzbromarone 0.6 μM； colchicine 0.9 μM；orotic acid 4.0 μM； oxypurinol 1.0 μM；probenecid 3.0 μM及 sulfinpyrazone 0.8 μM ( S/N=3，0.5 psi，注射5秒)。本法之準確度與精密度其R.S.D.及R.E. 均小於4 %，本法已成功應用於市售製劑allopurinol及colchicine之含量分析。

關鍵字

毛細管電泳法；抗痛風及降尿酸劑

並列摘要

Capillary electrophoresis method (CE) was studied for the simultaneous separation and quantitation of seven antigout and uricosuric drugs including allopurinol (AP), benzbromarone (BZB), orotic acid (OA), oxypurinol (OP), probenecid (PB) and sulfinpyrazone (SPZ). Capillary zone electrophoresis method (CZE) was first tried to the separation of the seven drugs. At the conditions of background electrolyte with borate buffer (50 mM; pH 9.00), applied voltage of 15 kV, untreated fused-silica capillary (40.2 cm, effective length 30 cm, 50 μm I.D.), hydrodynamic injection (0.5 psi, 5 s) and detection at 214 nm, CZE can separate and quantitate six drugs excluding colchicine with a quantitative range of 20-100 μM. Further application of micellar electrokinetic chromatography (MEKC) to the separation of the seven drugs was performed. The results indicate that MEKC can separate and quantities all the seven drugs including colchicine at the conditions of borate buffer (45 mM; pH 9.00) with 20 mM sodium dodecyl sulfate and other CE conditions similar to those used in CZE. On the method validation, the calibration curves were linear over a range of 5-200 μM for benzbromarone, sulfinpyrazone and colchicine and 10-200 μM for allopurinol, orotic acid, oxypurinol and probenecid, respectively (r > 0.999). The RSD and RE were all less than 4 % for the intraday and interday assay. All the recoveries were greater than 96 %. The limits of detection were 2.0 μM for allopurinol, 0.6 μM for benzbromarone, 0.9 μM for colchicine, 4.0 μM for orotic acid, 1.0 μM for oxypurinol, 3.0 μM for probenecid, and 0.8 μM for sulfinpyrazone (S/N=3, 0.5 psi, 5s). Partial application of the method to the determination of allopurinol and colchicine in commercial tablets proved feasible.

並列關鍵字

CZE ； Capillary electrophoresis ； antigout and uricosuric drugs ； MEKC

參考文獻

1 R.A. Terkeltaub, Gout, N. Engl. J. Med, 349, 17(2003) 1647-1655.

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