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首次提出超分子显色反应的概念。乳化剂OP存在下,在pH=2 8~3 2 的Clark Lubs缓冲介质中,2 (4 磺基苯偶氮) 1,8 二羟基3,6 萘二磺酸与血清蛋白质发生超分子显色反应形成红色超分子复合物,该超分子复合物的最大吸收波长为565 nm,而试剂本身最大吸收波长位于510nm,对比度为55 nm。详细研究了该超分子显色反应的条件,建立了测定多种蛋白质的分光光度新方法,不同蛋白质在10~130 mg/L至10~200 mg/L质量浓度范围内服从比尔定律,其表观摩尔吸光系数在1 774×104~5 882×105 L/(mol·cm)。所拟方法简便、快速、准确、灵敏,选择性高,重现性好,用于实际人血清样品中总蛋白含量的测定,结果与考马斯亮蓝G 250法一致。
For the first time put forward the concept of supramolecular chromogenic reaction. In the presence of emulsifier OP, 2 (4 sulfophenylazo) 1,8-dihydroxy-3,6-naphthalenedisulfonic acid and serum proteins undergo supra-molecular colorization in a Clark Lubs buffer at pH = 28-3 2 The reaction formed a red supramolecular complex with a maximum absorption wavelength of 565 nm and a maximum absorption wavelength of the reagent at 510 nm with a contrast ratio of 55 nm. The conditions of this supramolecular chromogenic reaction were studied in detail. A new spectrophotometric method for the determination of various proteins was established. Beer ’s law was obeyed by different proteins in the range of 10 ~ 130 mg / L to 10 ~ 200 mg / L, The molar extinction coefficient ranged from 1 774 × 104 to 5 882 × 105 L / (mol · cm). The proposed method is simple, rapid, accurate, sensitive, selective and reproducible. The proposed method was applied to the determination of total protein in real human serum samples. The results were consistent with the Coomassie Brilliant Blue G 250 method.