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.
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%.
出处
《兰州大学学报（自然科学版）》
CAS
CSCD
北大核心
2004年第3期38-42,共5页
Journal of Lanzhou University(Natural Sciences)
基金
国家自然科学基金资助项目(20275014)
关键词
三溴偶氮胂
共振光散射法
蛋白质
正交设计
arsenazo-TB
Rayleigh light scattering
proteins
orthogonal array design
