探针N-戊基-N′-(对氨基苯磺酸钠)硫脲快速测定血清样品中蛋白质含量的光谱法研究

A Rapid Method to Determine the Total Proteins in Serum Sample Studied by Synchronous Fluorescence Technique with N-(pentyl)-N′-(sodium p-aminobenzenesulfonate) thiourea as Probe

研究了N-戊基-N′-(对氨基苯磺酸钠)硫脲(APT)与人血清白蛋白(HSA)体系的三维荧光光谱,证明了两者可以发生相互作用,APT的加入导致HSA疏水微环境极性和构象的变化.考察了Δλ值,pH值、离子强度、试剂加入顺序及APT用量等因素对APT与HSA体系的同步荧光强度的影响.在最佳实验条件下,体系的同步荧光强度与HSA的浓度(2.0~479.0 mg.L-1范围内)呈良好的线性关系,线性相关系数为0.998,检出限可达1.3 mg.L-1.在此基础上,以APT为荧光探针,运用固定波长同步荧光光谱法测定了人血清样品中的蛋白质含量并进行了加标回收实验,回收率在102.8%~104.6%之间.结果显示,本方法简单、快速、灵敏度高、线性范围宽、体系稳定且具有较好的选择性和回收率.

The experiment is investigated for the three-dimensional spectra of N-(pentyl)-N′-(sodium p-aminobenzenesulfonate)thiourea(APT)-human serum albumin(HSA) system.The results show that APT can react with HSA,and the conformational and micro-environmental changes of HSA are investigated by comparing the three-dimensional spectral changes.The effect factors for spectral characterization and intensity of synchronous fluorescence such as the value of Δλ,pH,ionic strength,addition sequence,reagent concentration are investigated.Under the optimum conditions,the synchronous fluorescence intensity of the system is in proportion to the concentration of protein in the range of 2.0~479.0 mg·L-1 for human serum albumin(HSA),with the detection limit 1.3 mg·L-1 for HSA.The linear correlation coefficient is 0.998.It is used to determine protein in human serum sample,the recovery is within the range of 102.8%~104.6%.