蛋白质的曙红Y-中性红逆向能量转移荧光法测定  

Determination of Protein by Reverse Fluorescence Energy Transfer between Eosin Y and Neutral Red

作　　者：刘保生[1] 薛春丽[1] 王晶[1] 吕云开[1] 

机构地区：[1]河北大学理化分析中心药物化学与分子诊断教育部省部共建重点实验室,河北保定071002

出　　处：《分析测试学报》2009年第3期345-348,共4页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(20075005)

摘　　要：在pH2.35的B-R缓冲溶液中,酸性染料曙红Y(EY)与碱性染料中性红(NR)之间由于静电吸引能够发生有效的能量转移,使能量受体NR荧光增强,能量给体EY的荧光猝灭。在该体系中加入人血清白蛋白(HSA),由于竞争作用使得HSA的加入破坏了EY-NR间的能量转移,产生了明显的逆向反应,并且生成HSA-EY络合物,利用该络合物的荧光增加强度与HSA含量间的线性关系建立了一种测定微量血清蛋白的新方法。结果表明,血清蛋白工作曲线线性范围为0.6～12.0mg/L;方法检出限为0.25mg/L;6次平行测定相对标准偏差为1.94%～4.58%;回收率为96%～105%。该方法的稳定性好、选择性高,直接用于人血清试样中总蛋白含量的测定,实验结果与常用的双缩脲法基本一致。In the Britton - Robinson buffer solutions of pH 2.35, effective energy transfer could occur between Eosin Y（EY） and neutral red（NR） due to static attraction. The fluorescence intensity of EY was reduced while that of NR was increased. When human serum albumin（HSA） was added to this system, the fluorescence energy transfer between EY - NR pair was disturbed, causing reverse reaction to occur, resulting in the increase of fluorescence intensity of EY and the formation of HSA - EY complex. A new method for the determination of trace protein was established on the basis of the linear relationship between the fluorescence intensity of the coordinate compound and the concentration of HSA. The detection limit for this method was 0.25 mg/L. The determination range for HSA was 0. 6 - 12. 0 mg/L. Among six parallel experiments, the relative standard deviations were 1.94% - 4.58% and the recoveries were 96% - 105% . The method was used for the determination of proteins in human serum samples with good stability and high selectivity and the results obtained were in good agreement with those obtained by biuret spectrophotometric method.

关 键 词：曙红Y 中性红 荧光能量转移 蛋白 

分 类 号：O657.3[理学—分析化学] Q51[理学—化学]