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基于共振光散射增强用波尔多红测定纳克级蛋白质。

Determination of proteins at nanogram levels with Bordeaux red based on the enhancement of resonance light scattering.

作者信息

Feng Suling, Pan Zihong, Fan Jing

机构信息

College of Chemical and Environmental Sciences, Henan Normal University, Henan, Xinxiang 453007, PR China.

出版信息

Spectrochim Acta A Mol Biomol Spectrosc. 2006 Jun;64(3):574-9. doi: 10.1016/j.saa.2005.07.056. Epub 2006 Mar 10.

Abstract

A simple, sensitive and selective method was proposed for the determination of proteins by using a resonance light scattering technique. The weak resonance light scattering (RLS) of Bordeaux red (BR) can be enhanced greatly in the pH range 3.87-3.96 by the addition of micro amounts of proteins, resulting in four characteristic peaks in the wavelength range 250-600 nm. At the maximal wavelength of 363 nm, the enhanced RLS is proportional to the concentration of proteins in the range 0.12-10.8 microg ml-1 for bovine serum albumin (BSA) and 0.24-18.0 microg ml-1 for human serum albumin (HSA). The detection limits were 40.0 and 52.9 ng ml-1 for BSA and HSA, respectively. The present method has been applied to the determination of total proteins in human serum, urine and saliva samples. The obtained results are in good agreement with those obtained by the Bradford method with relative standard deviations (R.S.D.) of 0.9-2.3%.

摘要

提出了一种利用共振光散射技术测定蛋白质的简单、灵敏且具选择性的方法。通过加入微量蛋白质,在pH值3.87 - 3.96范围内,波尔多红(BR)的弱共振光散射(RLS)可大幅增强,在250 - 600 nm波长范围内产生四个特征峰。在最大波长363 nm处,对于牛血清白蛋白(BSA),增强的RLS与0.12 - 10.8 μg ml-1范围内的蛋白质浓度成正比,对于人血清白蛋白(HSA),则与0.24 - 18.0 μg ml-1范围内的蛋白质浓度成正比。BSA和HSA的检测限分别为40.0和52.9 ng ml-1。该方法已应用于人血清、尿液和唾液样本中总蛋白的测定。所得结果与采用Bradford法获得的结果高度一致,相对标准偏差(R.S.D.)为0.9 - 2.3%。

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