曙红Y-SDS体系共振瑞利散射法测定蛋白质

Determination of Protein with Eosin Y by Resonance Rayleigh Light Scattering Method Using SDS as Sensitizer

研究了曙红Y与蛋白质的结合反应。在十二烷基磺酸钠存在下及pH3.05的柠檬酸-NaOH介质中,蛋白质与曙红Y通过分子间作用力形成复合物,使最大波长365nm的共振光散射光谱得到加强,根据其共振光散射的增强程度,可用于蛋白质的定量测定。十二烷基磺酸钠的加入,使灵敏度提高2.6倍。牛血清白蛋白、γ-球蛋白的线性范围分别为0.05～3.7、0.10～5.6mg/L,检测限分别为12、18μg/L。用于人血清、牛奶、豆浆、尿液中总蛋白质的测定,结果与经典的考马斯亮蓝法一致。

The binding reactions of the dye Eosin Y with proteins have been studied. In the presence of SDS and in the citricacid-NaOH buffer medium (pH3.05), the Eosin Y combined with the proteins by intermolecular forces to form complexes,causing an enhancement of resonance light scattering (RLS) at maximum wavelength of 365nm. A novel method for the determinationof proteins based on the enhancement of RLS was developed. The sensitivity raisesd 2.6 times in the presence of SDS. The linearranges were 0.05～3.7 mg/L for the bovine albumin and 0.10~5.6 mg/L for human γ-IgG. The detection limits were 12 and 18μg/L, respectively .The method has been applied to determine proteins in human serum, milk, soybean milk and protein, samplein urine. The results obtained with this method were in accordance with the authentic brilliant blue method.