摘要
以NaCl为介质,在Tris-HCl缓冲液(pH 7.4)中,三维荧光光谱显示核苷类药物N-(6-嘌呤)-蛋氨酸(PYMBA)与人血清白蛋白可以发生相互作用。结果表明,PYMBA的加入使得人血清白蛋白内源荧光猝灭,且体系的同步荧光强度和溶液中人血清白蛋白的浓度呈良好的线性关系,基于此,建立了测定蛋白质的新方法。在选定的最佳实验条件(Δλ=50 nm,离子强度为0.1 mol/L,pH=7.4)下,同步荧光强度与人血清白蛋白在2.8～552 mg/L范围内的线性相关系数为0.9985。运用本方法测定人血清、唾液和尿液等生物样品中的蛋白质含量并进行加标回收实验,回收率在98.0%～103.1%之间。本方法的检出限可达0.1 mg/L。本方法所得结果与经典的考马斯亮蓝法一致。
In Tris-HCl and NaCl medium,the three-dimensional fluorescence spectra of HSA-2-（9H-purin-6-ylamino）-4-（methylthio）（PYMBA） system showed that the PYMBA can react with serum albumin and the endogenous fluorescence of human serum albumin was quenched.At the same time,the fluorescence intensity of the system was proportional to the concentration of serum albumin.Based on this,a synchronous fluorescence method for the determination of trace proteins was developed.The fluorescence intensity of the system was in direct proportion to the concentration of protein in the range from 2.8 to 552.0 mg/L,with the detection limit of 0.1 mg/L for HSA when Δλ=50 nm,Tris-HCl buffer（pH 7.4） and ionic strength was 0.1 mol/L of NaCl solution.This method was applied to determine the proteins in human serum,urine and saliva samples and the recovery was 98.0%-103.1%.The results obtained with this method coincided with those obtained by the coomassie brilliant blue method.The effects of coexistent substances were also investigated.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2011年第5期728-732,共5页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金(No.30970696)资助项目
