反相HPLC法快速测定人血清中抗坏血酸含量

KAPID DETERMINATION OF ASCORBIC ACID IN HUMAN SERUM BY REVBKSEDPHASE HPLC WITH ELECTROCHEMICAL DETECTION

应用ＵｌｔｒａｓｐｈｅｒｅＯｃｔｙｌ柱，建立了快速测定人血清中抗坏血酸含量的反相ＨＰＬＣ法。流动相为醋酸盐缓冲液（ｐＨ５．２５）。电化学检测器用碳膜电极和甘汞参比电极，工作电压＋０．６０Ｖ，灵敏度１００ｎＡ／Ｖ。抗坏血酸的平均回收率为１００．４６％，日内（ｎ＝５）和日间（ｎ＝８）的平均变异系数分别为３．８５％和６．８０％。最低检出浓度为０．１３６μｇ／ｍｌ，线性范围０．１３６μｇ／ｍｌ～１３．６μｇ／ｍｌ（ｎ＝９，ｒ＝０．９９９９）。

A reversed-phase HPLC method has been developed for rapid detec-tion of ascorbic acid in human serum using Ultrasphere Octyl column.A buffer contain-ing 0.02%EDTA-2K,0.54%sodium acetate and 0.0l%acetic acid(pH 5.25)wasused as mobile phase at a flow of 0.5ml/min. Detection of ascorbic acid was accom-plished by electrochemical detector using a carbon electrode and 232 calomel refereneelectrode. The applied potential was set at+0.60v and the sensitivity was l00nA/V.The mean absolute recovery was l00.46%. Within n day(n=5)and between day(n=8 )percision(cv)were 3.85%and 6.80%,respectively,The measurable lowest levelwas 0.136 μug/ml and the standard curve was linear over the concentration range of O.136μug/ml～13.60μug/ml(n=9,r=0.9999).