Siangproh Weena, Teshima Norio, Sakai Tadao, Katoh Shuji, Chailapakul Orawon
Department of Applied Chemistry, Aichi Institute of Technology, 1247 Yachigusa, Yakusa-cho, Toyota 470-0392, Japan.
Talanta. 2009 Sep 15;79(4):1111-7. doi: 10.1016/j.talanta.2008.12.068. Epub 2009 Jan 20.
A simple, automatic and practical system for successive determination of albumin and creatinine has been developed by combining sequential injection analysis (SIA) and highly sensitive dye-binding assays. Albumin detection was based on the increase in the absorbance due to complex formation between albumin and eosin Y in acidic media. The absorbance of the complex was monitored at 547 nm. For the creatinine assay, the concentration of creatinine was measured by reaction with alkaline picrate to form a colored product which absorbs at 500 nm. The influences of experimental variables such as effects of pH, reagent concentration, standard/sample volume and interferences were investigated. Under optimal conditions, the automated method showed linearity up to 20 mg L(-1) for albumin and 100 mg L(-1) for creatinine. The 3 sigma detection limits were 0.6 and 3.5 mg L(-1) for albumin and creatinine, respectively, and the relative standard deviations (n=10) were 2.49% for 20 mg L(-1) albumin, and 3.14% for 20 mg L(-1) creatinine. Application of the proposed method to the direct analysis of urinary samples yielded results which agreed with those obtained from the Bradford protein assay and a creatinine enzymatic assay according to a paired t-test. The results obtained should be a step towards developing a fully automated and reliable analytical system for clinical research, which requires direct determination of albumin and creatinine and/or its ratios.
通过结合顺序注射分析（SIA）和高灵敏度染料结合测定法，开发了一种用于连续测定白蛋白和肌酐的简单、自动且实用的系统。白蛋白检测基于在酸性介质中白蛋白与曙红Y形成复合物导致吸光度增加。在547nm处监测复合物的吸光度。对于肌酐测定，通过与碱性苦味酸盐反应形成在500nm处有吸收的有色产物来测量肌酐浓度。研究了诸如pH值、试剂浓度、标准品/样品体积和干扰等实验变量的影响。在最佳条件下，该自动化方法对白蛋白的线性范围高达20mg L(-1)，对肌酐的线性范围高达100mg L(-1)。白蛋白和肌酐的3σ检测限分别为0.6和3.5mg L(-1)，20mg L(-1)白蛋白的相对标准偏差（n = 10）为2.49%，20mg L(-1)肌酐的相对标准偏差为3.14%。将所提出的方法应用于尿液样品的直接分析，根据配对t检验，所得结果与通过Bradford蛋白质测定法和肌酐酶法获得的结果一致。所获得的结果应是朝着开发用于临床研究的全自动且可靠的分析系统迈出的一步，该系统需要直接测定白蛋白和肌酐及/或其比率。
