使用高效液相色谱法测定干血斑样本中的总谷胱甘肽
Determination of Total Glutathione in Dried Blood Spot Samples Using a High-Performance Liquid Chromatography.
作者信息
Kanďár Roman, Štramová Xenie, Drábková Petra, Brandtnerová Martina
机构信息
Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 532 10 Pardubice, Czech Republic
Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 532 10 Pardubice, Czech Republic.
出版信息
J Chromatogr Sci. 2015 Jul;53(6):879-85. doi: 10.1093/chromsci/bmu135. Epub 2014 Oct 25.
A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes.
描述了一种使用高效液相色谱-荧光检测法测定干血斑（DBS）样本中总谷胱甘肽（TGSH）的方法。从一组献血者中采集全血和DBS样本。用二硫苏糖醇还原谷胱甘肽并用乙醇沉淀蛋白质后，样本用萘-2,3-二甲醛衍生化以形成非常稳定、高荧光的衍生物。分离采用反相高效液相色谱法。乙醇和去离子水（8:92，v/v）的混合物用作流动相。该方法的分析性能令人满意：批内和批间变异系数均低于10%。加标DBS样本的定量回收率在98.3%至103.6%之间。所提出的方法成本低廉，适用于临床检测目的。
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