偶氮胂羧与蛋白质作用的光谱性质及其分析应用研究

Study on the spectral character of the reaction between carboxyarsenazo and proteins and its analytical applications

研究表明,在pH 1.5的Clark-Lubs(C-L)缓冲介质中,偶氮胂羧与多种蛋白质在室温下能迅速结合生成蓝色复合物,λmax为612nm,线性范围(HSA)为0～100mg/L,表观摩尔吸光系数为2.67×105L·mol-1·cm-1.研究了光谱性质及反应机理,试验了影响因素,在确定的最佳实验条件下,建立了以CAA为光谱探针的光度法测定人血清蛋白质的新方法.测定人血清样品中蛋白质的含量,与经典的考马斯亮蓝G-250法(CBB G-250)结果一致.

In pH 1.5 Clark-Lubs buffer, carboxyarsenazo could bind rapidly with many kinds of proteins at room temperature forming a blue complex, which had a maximum absorption wavelength at 612nm. Beer＇ s law was obeyed in the range from 0 to 100 rag/L（HSA）. The molar absorptivity of reaction product was 2.67 × 105L· mol^-1. cm^-1. The influencing factors and mechanism were studied. A new method for the determination of proteins by using carboxyarsenazo as a spectral probe was established under the optimal conditions. This method has been used for the determination of total protein in human serum samples. The results obtained agreed well with those by coomassie brilliant blue G-250 method.