反相高效液相色谱法安培检测器快速测定人血清中维生素C含量

Rapid Determination of Vitamin C in Human Serum by HPLC with Amperometric Detection

维生素C(下称VC)是维持人体健康的重要维生素之一。它广泛参与机体的氧化、还原和羟化等代谢过程,对维持机体正常生理功能具有重要作用。VC缺乏可导致坏血病和机体抗病能力降低。通过血清VC浓度测定,对坏血病的诊断和治疗,对于及时纠正VC缺乏及评价某些生理、病理条件下的机体状况具有重要的临床意义。VC的测定有紫外比色法,有HPLC-UV检测食品和体液中VC含量,但灵敏度低。还有反相离子对色谱法,而离子对试剂价格昂贵。本法具有简便、快速、灵敏等特点。

A rapld reversed-phase HPLC procedure with amperometrie detection which resolves and quantifies vitamin C in human serum is presented. A 4.6×200mm column packed with 5μm Ul trasphere C8 was used as tne analytical column and Ultrasphere ODS (4.6×45mm) was for the guard column. A buffer containing 0.02%EDTA-2K, 0.54% sodium acetate and 0.10% acetic acid at a pH of 5.25, was used as mobile phase at a flow rate of 0.5ml/min.Detection of vitamin C was accomplished by amperometry using a carbon electrode and 232 calomel reference electrode. The applied potential was set at +0.60V. The sensitivity is 50nA or 100nA. As low as 1.0-2.7ng vitamin C can be detected.