快速测定生物样品中蛋白含量的同步荧光法研究  

Rapid Method to Determine the Total Proteins in Biological Samples

作　　者：闫迎华[1] 杨艳杰[2] 万笑云[1] 崔凤灵[1] 张强斋[1] 党晓云[3] 渠桂荣[1] 

机构地区：[1]河南师范大学化学与环境科学学院,河南新乡453007 [2]漯河医学高等专科学校,河南漯河462002 [3]河南师范大学体育学院,河南新乡453007

出　　处：《河南师范大学学报（自然科学版）》2010年第2期108-111,共4页Journal of Henan Normal University(Natural Science Edition)

基　　金：国家自然科学基金(30970696);河南省基础与前沿技术研究计划项目(092300410239)

摘　　要：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.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.

关 键 词：胞苷酸(CMP) 人血清白蛋白(HSA) 考马斯亮蓝G-250(CBB) 同步荧光光谱 

分 类 号：O657.3[理学—分析化学]