Determination of protein with eriochrome black T as a probe of resonance light scattering

研究了金属指示剂铬黑T(EBT)作为共振光散射探针测定蛋白质的分析方法。实验表明 ,在pH =4 10的Britton Robinson缓冲溶液条件下 ,铬黑T只有极弱的光散射 ,它与蛋白质结合后有强烈的共振光散射作用。在λ =375nm处 ,光散射强度最大 ,光散射强度与蛋白质的浓度成正比。据此建立了一种测定蛋白质的新方法。该方法简便、快速、灵敏度高 ,对HSA的检出限达到 39μg/L ;线性范围为 0～ 15mg/L ,用于人体血清样品的分析并同考马斯亮蓝法比较 ,取得了令人满意的结果。同时亦研究了牛血清白蛋白 (BSA)、γ球蛋白、鸡蛋白蛋白、溶菌酶与染料EBT之间的作用。比较了 2...A new resonance light scattering (RLS) probe is presented. In the Britton-Robinson buffer at pH 4.10, eriochrome black T (EBT) combined with proteins by intermolecular forces, which resulted in resonance light scattering. The RLS intensity reaches maximum at 375 rim and is proportional to the concentration of protein. A novel method for the determination of micro-amount of protein was developed. The assay is simple, rapid and sensitive with, a detection limit of 39 mug/L and a linear range up to 15 mg/L. Ibis method was applied to the determination of human serum protein, and compared to Bradford method. The results are satisfactory. The interaction of EBT with bovine serum albumin (BSA), gamma-globulin, oval albumin, lysozyme was investigated. Two RLS from different fluorophotometers were compared, and the mechanism of RLS was also discussed.SCI(E)中国科学引文数据库(CSCD)606732-7353