Yasukawa Keiko, Shimosawa Tatsuo, Okubo Shigeo, Yatomi Yutaka
Department of Clinical Laboratory, The University of Tokyo Hospital, Tokyo, Japan.
Ann Clin Biochem. 2018 Jan;55(1):121-127. doi: 10.1177/0004563217693257. Epub 2017 Jan 23.
Background Human mercaptalbumin and human non-mercaptalbumin have been reported as markers for various pathological conditions, such as kidney and liver diseases. These markers play important roles in redox regulations throughout the body. Despite the recognition of these markers in various pathophysiologic conditions, the measurements of human mercaptalbumin and non-mercaptalbumin have not been popular because of the technical complexity and long measurement time of conventional methods. Methods Based on previous reports, we explored the optimal analytical conditions for a high-performance liquid chromatography method using an anion-exchange column packed with a hydrophilic polyvinyl alcohol gel. The method was then validated using performance tests as well as measurements of various patients' serum samples. Results We successfully established a reliable high-performance liquid chromatography method with an analytical time of only 12 min per test. The repeatability (within-day variability) and reproducibility (day-to-day variability) were 0.30% and 0.27% (CV), respectively. A very good correlation was obtained with the results of the conventional method. Conclusions A practical method for the clinical measurement of human mercaptalbumin and non-mercaptalbumin was established. This high-performance liquid chromatography method is expected to be a powerful tool enabling the expansion of clinical usefulness and ensuring the elucidation of the roles of albumin in redox reactions throughout the human body.
背景 人巯基白蛋白和人非巯基白蛋白已被报道为各种病理状况的标志物,如肾脏和肝脏疾病。这些标志物在全身的氧化还原调节中发挥重要作用。尽管在各种病理生理状况下已认识到这些标志物,但由于传统方法技术复杂且测量时间长,人巯基白蛋白和非巯基白蛋白的测量并不普遍。方法 基于先前的报道,我们探索了使用填充有亲水性聚乙烯醇凝胶的阴离子交换柱的高效液相色谱法的最佳分析条件。然后使用性能测试以及各种患者血清样本的测量对该方法进行验证。结果 我们成功建立了一种可靠的高效液相色谱法,每次测试的分析时间仅为12分钟。重复性(日内变异性)和再现性(日间变异性)分别为0.30%和0.27%(CV)。与传统方法的结果具有很好的相关性。结论 建立了一种用于临床测量人巯基白蛋白和非巯基白蛋白的实用方法。这种高效液相色谱法有望成为一种强大的工具,能够扩展临床应用,并确保阐明白蛋白在人体全身氧化还原反应中的作用。