探针5-硝基水杨酸同步荧光法测定生物样品中蛋白质的应用研究

在模拟人体生理条件下，基于5-硝基水杨酸与人血清白蛋白(HSA)相互作用生成复合物，导致血清白蛋白的内源荧光产生特异性变化，而建立了以5-硝基水杨酸为分子探针，用固定波长同步荧光光谱分析测定蛋白质的新方法。体系同步荧光光谱特征及强度受Δλ、反应介质、反应温度等因素的影响。对上述实验条件进行了优化，结果表明，在最佳实验条件下，体系的同步荧光强度(ISF)与人血清白蛋白在2.21 ~ 469.2 mg/L 的浓度范围内呈良好的线性关系，检测限为0.81 mg/L(n=11)。本实验对血清、尿样和唾液样品进行了测定，回收率在98.4% ~ 102.6 %之间。

Studied on the determination of proteins in biology samples by synchronous fluorescence technique with 2-Hydroxy-5-nitrobenzoic acid as a probe

2-Hydroxy-5-nitrobenzoic acid can react with serum albumin to form a complex. Based on this, a new method for the determination of proteins by synchronous fluorescence spectra under simulated physiological conditions was established using 2-Hydroxy-5-nitrobenzoic acid as a fluorescence probe. The spectral characterization and intensity of synchronous fluorescence were related to the value of Δλ, reaction medium and reaction temperature, and so on. On basis of these experimental results show that, under the optimum conditions, the synchronous fluorescence intensity of the system is in proportion to the concentration of protein in the range of 2.21 ~ 469.2 μg/mL for human serum albumin (HSA). The detection limit was 0.81 μg/mL (n=11) for HSA. On basis of these it had been applied to the determination of proteins in human serum, urine and saliva samples with satisfactory results, the recovery was within the range of 97.1~102.7%. Results suggested that the method possessed easy of implementation, rapidity, high sensitivity, broad linear range, and better RSD and recovery, and so on.