固定波长同步荧光法测定生物样品中蛋白含量

Determination of Proteins in Biological Samples by Fixed Wavelength Synchronous Fluorescence Spectrometry

不同代喹诺酮类药物分子与人血清白蛋白(HSA)相互结合,体系的同步荧光强度和溶液中HSA浓度呈良好的线性关系,其中氧氟沙星(OFLX)水溶性好,且对HSA的内源荧光有较强的猝灭作用,因而建立了以OFLX为分子探针,运用固定波长同步荧光法测定生物样品中蛋白质总含量的新方法。考察了△λ、介质、pH值、试剂用量和离子强度等因素对体系同步荧光的影响。在最佳实验条件下,体系同步荧光强度与HSA浓度在6.0×10-8～3.4×10-6 mol.L-1范围内的线性相关系数为0.9997。运用该方法对人血清、唾液和尿液等样品进行测定并进行加标回收实验,回收率在99.3%～103.5%之间。该方法具有简单、快速、准确度高、重现性好等优点,可直接用于血清、唾液和尿样等生物样品中蛋白质总量的快速测定。

Different kinds of quinolones can interact with serum albumin （HSA） and synchronous fluorescence intensities of these systems have good linear relative with serum albumin＇s concentration. Ofloxacin（OFLX） was chosen as the best molecular probe due to better water solubility and higher quenching efficiency among them. And fixed wavelength synchronous fluorescence spectrometry for the determination of proteins in biological samples was developed. Various factors including △λ, medium, reagent concentration and ionic strength were investagated. The linear range is 6.0 × 10^-8 - 3.4 × 10^-6 mol. L- 1 ,with a correlation coefficient 0. 9997. Human serum,saliva and urine were determined by this method and the recovery rates were 99.3%-103.5% . The results showed that the method was simple and rapid with high selectivity and good repeatability.