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在pH3.5左右的柠檬酸-NaOH介质中,蛋白质与铬天青S可以通过以静电引力为主的分子间作用力结合生成大分子离子缔合物,产生最大散射波长约为370nm的弹性光散射信号.基于这一现象可以测定低至0.02μg·mL~(-1)的血清白蛋白,工作曲线在0～1.0μg·mL~(-1)范围内呈线性关系.这一新方法的灵敏度比Coomassie亮蓝法高50倍.用普通的荧光分光光度计测量了这一体系的散射光谱,研究了pH,离子强度和试剂浓度等实验条件对散射光强的影响,考察了阳离子、阴离子和非离子表面活性剂以及氨基酸和常见金属离子对反应体系的作用.将这一方法用于测定尿中徽量蛋白,结果满意.
In citric acid-NaOH media at pH 3.5, the protein and chrome azure S can generate macromolecular ion association by intermolecular force mainly based on electrostatic attraction, resulting in elastic light with the maximum scattering wavelength of about 370 nm Scattering signal.According to this phenomenon, the serum albumin can be measured as low as 0.02μg · mL -1, and the working curve is linear in the range of 0 ~ 1.0μg · mL -1. The sensitivity was 50 times higher than that of Coomassie Brilliant Blue method.The scattering spectra of this system were measured by a common fluorescence spectrophotometer.The effects of pH, ionic strength and reagent concentration on the light intensity were studied. The effects of cation, anion And non-ionic surfactants as well as amino acids and common metal ions on the reaction system.This method was used to determine the urine protein in Anhui, the results were satisfactory.