快速测定生物样品中蛋白含量的同步荧光法研究

基于胞苷酸(CMP)与人血清白蛋白(HSA)相互作用,体系的同步荧光强度和溶液中白蛋白的浓度呈良好的线性关系,建立了以胞苷酸为分子探针,运用同步荧光光谱法测定生物样品中蛋白质含量的新方法.考察了Δλ值、介质、试剂用量、离子强度、加入顺序、反应时间等因素对体系同步荧光光谱的影响,在最佳实验条件下,体系的同步荧光强度与白蛋白在2.76～552.0μg·mL-1范围内的线性相关系数为0.9984,方法的检测限可达0.13μg·mL-1.用本方法对人血清、唾液和尿液等样品进行测定并做了加标回收实验,回收率在97.8%～103.6%之间.对11份空白溶液进行平行测定,相对标准差为1.16%.并用CBB法做了对照实验,取得了令人满意的结果.

Rapid Method to Determine the Total Proteins in Biological Samples

Cytidine-5'-monophosphate (CMP) can interact with serum albumin to form a complex and the synchronous fluorescence intensity of system has a good linear relative with the serum albumin concentration in the solution. Based on this,a new method for the determination of proteins in biological samples is developed utilizing CMP as a molecular probe. The factors of determination such as Δλ,medium,reagent concentration,ionic strength,addition sequence,reaction time,are investagated. The range is 2.76～552.0 μg·mL-1 and the detection limit is 0.13?g/mL Under the optimum conditions. Human serum,saliva and urine are determined by this method and standard addition experiment is done. The recovery rate is 97.8% ~ 103.6%. 11 blank solutions to the parallel experimental data obtain the relative standard deviation of 1.16%. CBB G-250 method is done as the control experiment and the result is in good accordance with synchronous fluorescence method.