Determination of uroporphyrin and coproporphyrin in urine by capillary electrophoresis

• Main Authors: Yu-Chen Huang, 黃俞甄
• Other Authors: Shou-Mei Wu
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/81276884956400151751

碩士 === 高雄醫學大學 === 藥學研究所 === 100 === In this study, a method of anion-selective exhaustive injection and sweeping micellar electrokinetic chromatography was established for determination of uroporphyrin dihydrochloride (U) and coproporphyrin dihydrochloride (C) in human urine. After alkalization and dilution, the samples were loaded by electrokinetic injection (-10 kV, 200 s). The capillary is filled with phosphate buffer (70 mM, pH 3), and then with a water plug (2.5 psi, 10 sec) before anion-selective exhaustive injection. The separation was performed at -20 kV and detected at 400 nm using phosphate buffer (70 mM, pH 3) containing 100 mM sodium dodecyl sulfate. During method validation, calibration curves were linear over a range of 15-200 ng/ml for U, and 300-1000 ng/ml for C. The correlation coefficients for the calibration curves were greater than 0.983 for the intra- and interday analysis. The RSD and RE in intra- and interday were all less than 8.6%. The LODs (S/N = 5) were 5 ng/ml for U, and 100 ng/ml for C. This analysis can be used to diagnose porphyrias.