光谱法研究N-丁基-N'-(对氨基苯磺酸钠)硫脲与人血清白蛋白的相互作用及其分析应用

The interaction of N-(n-butyl)-N′-(sodium p-aminobenzenesulfonate)thiourea with human serum albumin and its analytical application

利用荧光光谱法和紫外光谱法研究了模拟生理条件下N-丁基-N'-(对氨基苯磺酸钠)硫脲(BPT)与人血清白蛋白(HSA)的相互作用。研究结果表明,BPT对HSA内源荧光的猝灭是静态猝灭。由热力学参数确定了BPT与HSA间存在疏水作用,根据Frster能量转移理论计算出结合距离为2.97 nm。通过三维荧光光谱研究了BPT对HSA构象的影响。在二者相互作用的基础上,以BPT为分子探针,运用同步荧光光谱法测定了生物样品中的蛋白质含量。对人血清、唾液和尿样进行平行测定并进行加标回收实验,回收率在96.2%～101.9%之间。

This study was calculated to examine the interaction of N-（n-butyl）-N′-（sodium p-aminobenzenesulfonate）thiourea（BPT） with human serum albumin（HSA） by fluorescence spectroscopy and ultraviolet spectroscopy under the condition of physiology.The results showed that BPT had a strong ability to quench the intrinsic fluorescence of HSA through static quenching procedure.The intermolecular forces may be the hydrophobic interaction according to the thermodynamic parameters.In terms of the Frster theory of non-radiation energy transfer,the binding distances was obtained.The three-dimensional fluorescence spectrum was applied to investigation of the conformation of HSA with BPT.Moreover,on the basis of the interaction of BPT with HSA the synchronous fluorescence technique was successfully employed to determine the total proteins in biological samples.This method was used for the determination of the proteins in human serum,saliva and urine samples and the recoveries were 96.2%~101.9%.