Cui Feng-ling, Yan Ying-hua, Zhang Qiang-zhai, Qu Gui-rong, Lu Yan
School of Chemistry and Environmental Science, Henan Normal University, Xinxiang 453007, China.
Guang Pu Xue Yu Guang Pu Fen Xi. 2009 Sep;29(9):2531-4.
The authors studied the characterization of synchronous fluorescence spectra of 5-methyluridine-protein system; the spectral characterization and intensity of synchronous fluorescence were related to the value of deltalamda, reaction medium, reagent concentration, ionic strength, addition sequence, reaction time, reaction temperature, and so on. On basis of the experimental results, the new method for the determination of the proteins was developed with 5-methyluridine as a molecular probe. Under the optimum experimental conditions, the synchronous fluorescence intensities of 5-methyluridine-HSA systems were in good proportion to the HSA concentration of the system in the range of 1.38-575.2 microg x mL(-1) and the detection limit could achieve 0.12 microg x mL(-1). The method is simple and rapid. Biological samples such as human serum, saliva and urine were determined utilizing this method, standard addition experiment was done, and the recovery rate was 98.7%-103.8%. Eleven blank solutions were used for the parallel experiment, resulting in a relative standard deviation of 1.56%. The results show that the method using synchronous fluorescence spectroscopy with 5-methyluridine as a molecular probe is simple, rapid and highly sensitive with a wide linear range, good stability and high selectivity. The method was applied directly to determine the total proteins in human serum, saliva and urine samples, and the results were satisfactory.
作者研究了5-甲基尿苷-蛋白质体系同步荧光光谱的表征；同步荧光的光谱特征和强度与Δλ值、反应介质、试剂浓度、离子强度、添加顺序、反应时间、反应温度等有关。基于实验结果，开发了以5-甲基尿苷为分子探针测定蛋白质的新方法。在最佳实验条件下，5-甲基尿苷-HSA体系的同步荧光强度与体系中HSA浓度在1.38 - 575.2 μg·mL⁻¹范围内呈良好比例关系，检测限可达0.12 μg·mL⁻¹。该方法简便快速。利用该方法对人血清、唾液和尿液等生物样品进行了测定，进行了标准加入实验，回收率为98.7% - 103.8%。采用11个空白溶液进行平行实验，相对标准偏差为1.56%。结果表明，以5-甲基尿苷为分子探针的同步荧光光谱法简便、快速、灵敏度高，线性范围宽，稳定性好，选择性高。该方法直接应用于人血清、唾液和尿液样品中总蛋白的测定，结果令人满意。
