应用化学 ›› 2011, Vol. 28 ›› Issue (07): 849-854.DOI: 10.3724/SP.J.1095.2011.00446

• 研究论文 • 上一篇    

白藜芦醇探针同步荧光法测定蛋白质含量

邢卫卫,崔凤灵*,毛润泽,程姗,霍瑞娜   

  1. (河南师范大学化学与环境科学学院 新乡 453007)
  • 收稿日期:2010-08-02 修回日期:2010-10-30 出版日期:2011-07-10 发布日期:2011-07-10
  • 通讯作者: 崔凤灵,教授; Tel:0373-3326335; Fax:0373-3326336; E-mail:fenglingcui@hotmail.com; 研究方向:荧光分析
  • 基金资助:
    国家自然科学基金资助项目(30970696 )

Synchronous Fluorescence Spectrometric Determination of Proteins with Resveratrol as Probe

XING Weiwei, CUI Fengling*, MAO Runze, CHENG Shan, HUO Ruina   

  1. (School of Chemistry and Environmental Science,Henan Normal University,Xinxiang 453007)
  • Received:2010-08-02 Revised:2010-10-30 Published:2011-07-10 Online:2011-07-10
  • Contact: CUI Fengling

摘要:

在Tris-HCl缓冲溶液体系中(pH=7.4),白藜芦醇与人血清白蛋白相互作用,对蛋白的内源性荧光产生猝灭作用,而且,同步荧光的强度与人血清白蛋白的浓度成正比。 基于此,建立了以白藜芦醇为荧光探针,运用固定波长同步荧光光谱法测定生物样品中蛋白质含量的新方法。 体系同步荧光光谱特征及强度受Δλ、反应介质和离子强度等因素的影响。 在最佳实验条件下,体系的同步荧光强度(I)与人血清白蛋白在1.380~276.0 mg/L的浓度范围内呈良好的线性关系,检测限为1.037 mg/L(n=11)。 对血清、尿样和唾液等生物样品进行测定,回收率在93.5%~105.8%之间,与传统的考马斯亮蓝(G-250)法作对照,结果一致。

关键词: 白藜芦醇, 人血清白蛋白, 同步荧光光谱, 考马斯亮蓝, 三维荧光

Abstract:

The threedimensional spectra combined with fluorescence spectroscopy under simulative physiological conditions were used to investigate the interaction between resveratrol and human serum albumin. With the presence of Tris-HCl, the endogenous fluorescence was quenched by resveratrol and the three-dimensional spectra also changed when resveratrol exsited in the system. The synchronous fluorescence intensity and the concentration of serum albumin solution have a good linear relationship. A new synchronous fluorescence method for the determination of trace proteins was developed. The spectral characterization and intensity of synchronous fluorescence were related to the value of Δλ, reaction medium and so on. Under optimized conditions, the fluorescence intensity(I) of the system was proportional to the concentration of protein in the range of 1.380~276.0 mg/L, with the detection limit of 1.037 mg/L for human serum albumin(HSA). This method was applied to the determination of proteins in human serum, urine and saliva samples and the recovery was 93.5%~105.8%. The results obtained with this method were accordant with those obtained by the coomassie brilliant blue(G-250) method.

Key words: resveratrol, human serum albumin, synchronous fluorescence spectra, coomassie brilliant blue, three-dimensional fluorescence

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