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三溴偶氮胂共振光散射法定量测定蛋白质

Quantitative determination of proteins by Rayleigh light scattering after the optimization of the reaction with arsenazo-TB

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【作者】 贾润萍陈晓峰董立军李前锋陈兴国胡之德

【Author】 JIA Run-ping, CHEN Xiao-feng, DONG Li-jun, LI Qian-feng, CHEN Xing-guo, HU Zhi-de(College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, China)

【机构】 兰州大学化学化工学院兰州大学化学化工学院 甘肃 兰州 730000甘肃 兰州 730000甘肃 兰州 730000

【摘要】 首次研究了三溴偶氮胂和蛋白质作用的共振光散射光谱.实验发现,在pH 3.5-4.7的范围内,蛋白质的加入导致三溴偶氮胂共振光散射增强,并在345和410 nm处产生两个增强峰,其强度与蛋白质的浓度成线性关系,据此建立了一种定量测定蛋白质的共振光散射新方法.采用五水平正交设计对反应参数进行选择.方法的线性范围为2.5-30.0μg/mL(BSA)和2.5-40.0μg/mL(HSA),对应的检测限分别为46.5和43.8 ng/mL.该方法已用于实际样品中蛋白质含量的测定,结果令人满意.

【Abstract】 A simple, sensitive and selective method has been developed for the determination of proteins using Rayleigh light scattering (RLS) technique and the method is based on the interaction of protein and Arsenazo-TB (TEA) at pH 4.1, which causes a substantial enhancement of the light scattering signal of Arsenazo-TB in the wavelength range of 300-550 nm with the maximal RLS enhancement at 345 and 410 nm. The reaction parameters, such as acidity, volume of buffer solution and concentration of Arsenazo-TB, were evaluated by the 5-level orthogonal array design(OAD). With this method, 2.5-30.0 μg/mL of bovine serum albumin(BSA) and 2.5-40.0 μg/mL of human serum albumin(HSA) can be determined and the detection limits, calculated three times with the standard deviation of six blank measurements, were 46.5 ng/mL and 43.8 ng/m for BSA and HSA respectively. Total proteins in human serum samples were determined; the relative standard deviation was no more than 5% and the recovery was 99.2%-100.6%.

【基金】 国家自然科学基金资助项目(20275014)
  • 【文献出处】 兰州大学学报 ,Journal of Lanzhou University , 编辑部邮箱 ,2004年03期
  • 【分类号】O657.32
  • 【被引频次】16
  • 【下载频次】102
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