变色酸2R共振光散射光谱法测定蛋白质

Determination of Protein with Chromotrope 2R by Resonance Light-scattering Method

基于人血清白蛋白 (HSA)对酸性偶氮染料变色酸 2R的共振光散射的增强效应 ,拟定了一种新的测定HSA的共振光散射法 .在pH =4.10的条件下 ,变色酸 2R本身只有极弱的光散射 ,但它与蛋白质的结合物却有强烈的共振光散射作用 ,在λex=λem =42 0nm处 ,光散射有最大的散射强度 ,并且光散射强度与蛋白质HSA的浓度成线性关系 ,线性范围为 0 .0～ 10 .0 μg mL ,检测限可达 0 .12 μg mL .该法简便、快速 ,用于人体血清样品蛋白质的测定并与考马斯亮蓝法比较 。

A new RLS (Resonance light scattering) method is presented in this paper. In the buffer of pH=4.10, chromotrope 2R is combined with proteins by intermolecular forces, which results in an enhanced resonance light scattering. The RLS intensity at 420 nm is maximum and proportional to the concentration of protein. The method is simple, rapid and sensitive. The detection limit is 0.12 μg/mL and the linear range is 10.0 μg/mL. It has been applied to the determination of human serum sample with satisfactory results.