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在pH2.6的Clark-Lubs缓冲介质中,紫红色的偶氮硝羧λmax为545 nm,与金属Sr(Ⅱ)生成稳定的β型蓝色配合物,其λmax为717 nm,红移172 nm。蛋白质(HSA)能与该配合物形成紫红色三元复合物,发生消光反应,吸光度呈对应波长下的负吸收。实验确定了反应的最佳条件,测定了有可能干扰的物质,探讨其β型配合物对不同蛋白质的响应情况,发现胃蛋白酶有较大灵敏度,ε=5.16×105L·mol-1·cm-1,在10～50μg/mL范围内遵循比尔定律。建立了以偶氮硝羧-Sr(Ⅱ)β型配合物与蛋白质发生褪色反应测定蛋白质含量的光度分析新方法,用于胃液中蛋白酶的测定,结果和经典方法吻合。
In the Clark-Lubs buffer medium with pH2.6, the violet-colored azo nitrate has a λmax of 545 nm and forms a stable β-type blue complex with metal Sr (Ⅱ) with a λmax of 717 nm and a redshift of 172 nm . Protein (HSA) can form a fuchsine ternary complex with the complex, extinction occurs, and the absorbance is negatively absorbed at the corresponding wavelength. The optimum reaction conditions were determined experimentally, the possible substances were determined and the response of different β-type complexes to different proteins was investigated. The results showed that pepsin had higher sensitivity, ε = 5.16 × 105L · mol-1 · cm- 1, follow Beer’s law in the range of 10 ~ 50μg / mL. A new spectrophotometric method was developed for the determination of protein content by the fading reaction of the azo-nitro-carboxyl-Sr (Ⅱ) complex with protein, which was used for the determination of protease in gastric juice. The results were in good agreement with the classical methods.