Ueyama Jun, Ishikawa Yusuke, Kondo Takaaki, Motoyama Megumi, Matsumoto Hiroyuki, Matsushita Tadashi
Department of Pathophysiological Laboratory Sciences, Field of Radiological and Medical Laboratory Sciences, Nagoya University Graduate School of Medicine, Nagoya, Japan
Department of Medical Technique, Nagoya University Hospital, Nagoya, Japan.
Ann Clin Biochem. 2015 Jan;52(Pt 1):144-50. doi: 10.1177/0004563214531930. Epub 2014 Mar 27.
Previously, high-performance liquid chromatography (HPLC) equipped with ultraviolet or fluorescence detectors has been used for separation of human mercaptalbumin (HMA) and human non-mercaptalbumin (HNA). However, it is difficult to perform reliable chromatographic analysis due to peak interference of such serum compounds as uric acid and bilirubin. The aim of this study is to explore a selective and simple analytical method for the determination of HMA and HNA.
HMA and HNA in serum sample were separated by HPLC and reacted with bromocresol green using a postcolumn reaction scheme.
A complete separation of HMA and HNA is achieved in less than 30 min by using weak anion exchange columns and isocratic elution. Within-run and between-day precisions at albumin concentration of 45 g/L were 4.2 and 1.7% for HMA and 4.5 and 4.6% for HNA, respectively. There was no interference in HMA and HNA peaks when bilirubin-, haemoglobin- or chyle-spiked pooled serum samples were analysed.
Our method is reliable and not labour-intensive and, therefore, might be applicable for clinical and epidemiological studies.
此前，配备紫外或荧光检测器的高效液相色谱法（HPLC）已用于分离人巯基白蛋白（HMA）和人非巯基白蛋白（HNA）。然而，由于尿酸和胆红素等血清化合物的峰干扰，难以进行可靠的色谱分析。本研究的目的是探索一种选择性且简便的分析方法来测定HMA和HNA。
血清样本中的HMA和HNA通过HPLC分离，并采用柱后反应方案与溴甲酚绿反应。
使用弱阴离子交换柱和等度洗脱，在不到30分钟内实现了HMA和HNA的完全分离。白蛋白浓度为45 g/L时，HMA的批内精密度和日间精密度分别为4.2%和1.7%，HNA分别为4.5%和4.6%。分析胆红素、血红蛋白或乳糜加标的混合血清样本时，HMA和HNA峰无干扰。
我们的方法可靠且不耗费人力，因此可能适用于临床和流行病学研究。
