亮绿SF(淡黄)与蛋白质相互作用的光度法研究及其分析应用

Spectrophotometric study on interaction of light green SF(yellowish) with protein and its analytical application

在pH 2.0的B-R缓冲介质中,蛋白质与亮绿SF(淡黄)(LGSF)在室温下结合生成复合物,其最大吸收波长为665 nm。本文采用光度法研究了结合反应的最佳条件、结合常数及最大结合位点数,并在此基础上建立了一个测定蛋白质的新方法。该方法测定牛血清蛋白(BSA)、人血清蛋白(HSA)及γ-球蛋白G(IgG)的线性范围至少可达80μg/mL,其表观摩尔吸光系数ε分别为:7.25×105、7.22×105与1.84×106L.mol-1.cm-1。除阴阳离子表面活性剂外,其余大部分物质不干扰蛋白质测定。可见方法具有选择性好、灵敏度高等优点。用于人血清样品中总蛋白的测定,所得结果与考马斯亮蓝G-250法基本一致。

In the britton-robison medium with pH 2.0, light green SF （yellowish） can bind with protein at room temperature to form a complex, which has a maximum absorption wavelength at 665 nm. The optimum reaction conditions, binding constant and maximum binding number of light green SF to proteins were investigated by spectrophotometry, and a new method for the determination of proteins was developed based on this binding reaction. The linear range for the determination of bovine serum albumin （BSA）, human serum albumin （HAS）, and Human Immunoglobulin G （IgG） can reach more than 80 tsg/mL with the apparent molar absorptivity of 7.25×10^5, 7.22×10^5 and 1.84×10^6 L·mol^-1·cm^-1, respectively. A lot of foreign substances do not interfere with the determination of proteins except for the anionic surfactants and cationic surfactants. The results of the determination for the total proteins in human serum sampies obtained by this new proposed method, which is of good selectivity and high sensitivity, agree well with those obtained by the coomassie brillant blue G-250 method.