应用高效液相色谱结合电化学检测技术建立了尿毒症病人血浆及透析液中抗坏血酸(AA)及脱氢抗坏血酸(DHAA)的测定方法.采用0.8 g/L偏磷酸与18%(体积分数)高氯酸的混合溶液作为生物样本中的蛋白沉淀剂及AA的提取剂,可大大提高AA在血浆中的稳定性.该方法已成功地应用于临床维生素C的常规检验.AA浓度为2 μmol/L～40 μmol/L时,其峰面积与浓度的相关系数大于0.99.AA与DHAA的日内相对标准偏差(RSD)分别小于8.9%和10.5%.AA在血浆及透析液中的回收率分别大于95%和78%.
A convenient and valid method for the determination of ascorbic acid(AA) and dehydroascorbic acid(DHAA) in plasma and dialysate from patients with uremia by high performance liquid chromatography with electrochemical detection is described. A mixture of 0.8 g/L metaphosphoric acid and 18%(volume fraction) perchloric acid was used as a protein precipitant and the extractant for AA from biosamples. It was also a good stabilizer for AA in samples. The proposed method is satisfied for routine screening of vitamin C in clinical applications with a correlation coefficient of more than 0.99 in the range of 2 μmol/L-40 μmol/L AA. The within-day precision was less than 8.9% and 10.55% for AA and DHAA, respectively. The recoveries of AA in plasma and dialysate were over 95% and 78%, respectively.
参考文献
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