【摘要】 目的应用高效液相色谱(high performance liquid chromatography,HPLC)和电化学检测技术建立快速测定慢性肾衰竭(chronic renal failure,CRF)患者血浆抗坏血酸(ascorbic acid,AA)及脱氢抗坏血酸(dehydroascorbic acid,DHAA)含量的方法。方法应用15%的偏磷酸溶液作为血液样本的蛋白沉淀剂及抗坏血酸的保护剂,处理后血浆样品-70℃保存,可极大提高血样中抗坏血酸的稳定性;分析柱为Inertsil ODS-3(DIKMA)150mm×4.6 mm,4.5μm;进样量为10μl,流速为0.8 ml/min,电压为0.8 V,柱温25℃;流动相由50 mmol/L磷酸钾和1 mmol/L乙二胺四乙酸(EDTA)溶液组成,pH为3.0。结果线性范围2μmol/L至200μmol/L;其峰高与浓度的相关系数大于0.999;抗坏血酸的平均回收率为101.89%,AA的日内(n=5)和日间(n=5)的平均相对标准偏差(RSD)分别为1.710%和5.915%。结论用高效液相色谱和电化学检测法检测CRF患者血浆中AA及DHAA含量,特异性强,灵敏度高。更多还原

【Abstract】 Objective A improved and rapid method for determination of ascorbic acid(AA) and dehydroascorbic acid(DHAA) in plasma from chronic renal failure patients by high performance liquid chromatography(HPLC) with electrochemical detection was presented.Methods A 15%(g/L)metaphosphoric acid solution was used as precipitant and protectant for AA from blood samples.The processed plasma samples was stored at-70℃ untill use.It would greatly improve stability of AA in samples.The analytical column was a Inertsil ODS-3(DIKMA)150 mm×4.6 mm,4.5 μm.An aliquot of 10 μl analyte for the samples was injected into HPLC at a flow rate of 0.8 ml/min.The applied potential was set at 0.8 V.The mobile phase was consisted of 50mmol/L potassium phosphate buffer containing 1mmol/L ethylenediaminetetraacetic acid(EDTA) at pH3.0.Results The correlation coefficient was more than 0.999 between the height and concentration in the range of 2 μmol/L-200 μmol/L.The average recovery of AA in plasma was 101.89%.The average within-day precision(n=5)and between-day precision(n=5)of AA were 1.710% and 5.915%.Conclusion The method for determination of ascorbic acid(AA) and dehydroascorbic acid(DHAA) in plasma from chronic renal failure patients by high performance liquid chromatography(HPLC) with electrochemical detection is convenient,rapid,and sensitive.更多还原