同步荧光法测定生物样品中蛋白含量的研究  

Determination of Protein in Biological Samples by Synchronous Fluorescence

作　　者：崔凤灵[1] 闫迎华[1] 张强斋[1] 渠桂荣[1] 卢雁[1] 

机构地区：[1]河南师范大学化学与环境科学学院,河南新乡453007

出　　处：《光谱学与光谱分析》2009年第9期2531-2534,共4页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(20673034;20772024);河南省教育厅自然科学基金项目(2006150012);河南省新乡市科技攻关项目(06G086)资助

摘　　要：基于5一甲基尿苷（5一Methyluridine）与血清白蛋白（HSA）相互作用，导致血清白蛋白的同步荧光发生特异性变化，且体系的同步荧光强度和溶液中HSA的浓度呈良好的线性关系，建立了以5-甲基尿苷为分子探针，运用固定波长同步荧光光谱法测定生物样品中蛋白质总含量的新方法。文章考察了猷值、反应介质、试剂用量、离子浓度、试剂加入顺序、反应时间、反应温度等因素对体系同步荧光的影响。在选定的最佳实验条件下，体系的同步荧光强度与血清白蛋白在1．38～575．2μg·mL^-1范围内的线性相关系数为0．9981，方法的检测限可达0．12μg·mL^-1。运用本方法对人血清、唾液和尿液等生物样品进行测定并进行加标回收实验，回收率在98．7％～103．8％之间。对11份5一甲基尿苷空白溶液进行平行测定，其相对标准差为1．56％。还考察了一些常见离子和有机物存在时对蛋白质测定的影响。结果显示，本方法具有简单、快速、灵敏度较高、线性范围宽、体系稳定、精密度高、重现性好等优点。该法可直接用于血清、唾液和尿样等生物样品中蛋白质总量的快速测定，结果令人满意。The authors studied the characterization of synchronous fluorescence spectra of 5-methyluridine-protein system~ the spectral characterization and intensity of synchronous fluorescence were related to the value of △λ, reaction medium, reagent con- centration, ionic strength, addition sequence, reaction time, reaction temperature, and so on. On basis of the experimental results, the new method for the determination of the proteins was developed with 5-methyluridine as a molecular probe. Under the optimum experimental conditions, the synchronous fluorescence intensities of 5-methyluridine-HSA systems were in good pro- portion to the HSA concentration of the system in the range of 1.38-575.2μg·mL^-1and the detection limit could achieve 0. 12 μg·mL^-1. The method is simple and rapid. Biological samples such as human serum, saliva and urine were determined utilizing this method, standard addition experiment was done, and the recovery rate was 98. 7%-103.8%. Eleven blank solutions were used for the parallel experiment, resulting in a relative standard deviation of 1.56%. The results show that the method using synchronous fluorescence spectroscopy with 5-methyluridine as a molecular probe is simple, rapid and highly sensitive with a wide linear range, good stability and high selectivity. The method was applied directly to determine the total proteins in human serum, saliva and urine samples, and the results were satisfactory.

关 键 词：5-甲基尿苷(5-Methyluridine) 人血清白蛋白(HSA) 同步荧光光谱 分子探针 

分 类 号：O657.3[理学—分析化学]