8-溴鸟苷探针测定生物样品中蛋白质含量的光谱法研究

Studied on the determination of protein by spectroscopic methods with 8-bromoguanosine as probe

研究了8-溴鸟苷（BG）与人血清白蛋白（HSA）体系的荧光猝灭光谱、三维荧光光谱，证明了两者可以发生相互作用，导致人血清白蛋白疏水微环境极性的改变和构象的变化；还分析了△入值、pH值、离子强度等因素对该体系同步荧光强度的影响，选定了8-溴鸟苷测定生物样品中蛋白质含量的最佳实验条件；在最佳实验条件下，体系的同步荧光强度与人血清白蛋白在1．38～262．20μg／mL范围内呈良好的线性关系，线性相关系数为0．9994，检出限1．281μg／mL。以8一溴乌苷为荧光探针，运用固定波长同步荧光光谱法测定了人血清、唾液和尿液等生物样品中的蛋白质含量并进行了加标回收实验，回收率在95．O％～103．8％之间。本实验还用经典的考马斯亮蓝法（CBB）进行了对比，与同步荧光光谱法测定结果基本一致。

The experiment were investigated for the fluorescence spectra and the three-dimensional spectra of 8- bromoguanosine （BG）-human serum albumin （HSA） system. The results showed that BG can react with HSA, and the confonnational and micro-environmental changes of HSA were investigated by comparing the three- dimensional spectral changes in absence and presence of BG. The effect factors for spectral characterization and intensity of synchronous fluorescence such as the value of Ak, pH, ionic strength, and so on, were alsoinvestigated. And then the optimum conditions of this experiment were selected. The linear range of this synchronous fluorescence method was 1.38 - 262.2 μg/mL, with the detection limit 1. 281 μg/mL for HSA. The linear correlation coefficient was 0. 9994. This method was applied to the determination of the proteins in human serum, saliva and urine samples and the recovery was 95.0% - 103.8%. The Coomassie Brilliant Blue （CBB） method was also done as the control experiment and the results were in good accordance with those by synchronous fluorescence method.