Alternative method for measurement of albumin/creatinine ratio using spectrophotometric sequential injection analysis
Alternative method for measurement of albumin/creatinine ratio using spectrophotometric sequential injection analysis
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使用分光光度连续注射分析测量白蛋白/肌酐比率的替代方法
DOI：
10.1016/j.talanta.2008.12.068
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发表时间：
2009
期刊：
影响因子：
6.1
通讯作者：
O. Chailapakul
中科院分区：
文献类型：
--
作者：
W. Siangproh;N. Teshima;T. Sakai;S. Katoh;O. Chailapakul
将顺序注射分析(SIA)与高灵敏的染料结合分析相结合，建立了一种简单、自动、实用的白蛋白和肌酐连续测定系统。白蛋白的检测是基于白蛋白与曙红Y在酸性介质中形成络合物而导致的吸光度增加。络合物的吸光度在547 nm处被监测。肌酐测定是通过与碱性苦味酸盐反应生成有色产物，在500 nm处吸收来测定肌酐的浓度。考察了pH、试剂浓度、标样体积比、干扰等实验参数对测定结果的影响。在最佳条件下，白蛋白和肌酐的线性范围分别为20mgL−1和100mgL−1。3种σ方法对白蛋白和肌酐的检出限分别为0.6mgL−-1和3.5mgL-−-1，相对标准偏差(n=10)分别为2.49%和3.14%。将该方法应用于尿样的直接分析，得到的结果与Bradford蛋白测定法和肌酸酶测定法的结果一致。所获得的结果应该是朝着开发全自动和可靠的临床研究分析系统的方向迈出的一步，这需要直接测定白蛋白和肌酐和/或其比率。
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 547nm. For the creatinine assay, the concentration of creatinine was measured by reaction with alkaline picrate to form a colored product which absorbs at 500nm. 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 20mgL−1for albumin and 100mgL−1for creatinine. The 3σ detection limits were 0.6 and 3.5mgL−1for albumin and creatinine, respectively, and the relative standard deviations (n=10) were 2.49% for 20mgL−1albumin, and 3.14% for 20mgL−1creatinine. 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.