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Development of capillary electrophoretic technique for analyzing nucleosides in urine

 

Development of capillary electrophoretic technique for analyzing nucleosides in urine

年度:

101

專題生:

黃玟玲鍾均孟

吳佳憲

指導老師:

胡焯淳

邱泰嘉

查詢關鍵字:

 Cancer biomarker; Capillary electrophoresis, Creatinin;, Nucleosides; Urine

摘要:

 

   We developed a simple method for rapid detection of four urinary nucleosides, creatinine, and uric acid by capillary electrophoresis (CE) coupled with UV detection. CE was performed in a capillary of 60 cm × 75 μm I.D. (effective length, 50 cm). The optimum buffer system for the separation of nucleosides in micellar electrokinetic chromatography was 15 mM borate buffer (pH 9.76) with 25 mM sodium dodecyl sulfate. The UV detector was set at 254 nm and the applied voltage was 16 kV, the separation was completed within 10 min. The LODs were ranged from 0.17μM to 0.36μM. In addition, we analyze the urinary creatinine and uric acid by capillary zone electrophoresis. The optimized conditions were using 20 mM phosphate buffer (pH 3). The UV detector was set at 240 nm and the applied voltage was 25 kV, the separation was completed within 10 min. The LODs (S/N = 3) were 0.83 μM and 1.99 μM for uric acid and creatinine, respectively. The RSD of intraday and interday of both CE methods were less than 4%. Comparison the urine samples from cancer patients and healthy people, urinary cytidine and uridine level were 576-fold and 18-fold higher cancer patients and than in healthy people. These two nucleosides are potential cancer biomarkers, and can be applied in study of urinary cancer biomarkers.

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