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在pH3.94的HAc-NaAc缓冲介质中,偶氮胂羧与Cu2+反应生成稳定的紫红色配合物,该配合物与生物大分子蛋白质发生染色反应形成复合物,λmax646nm,比试剂本身的541nm红移105nm,ε646nm=2.10×105L/mol·cm-1(BSA),蛋白质在20～100μg/mL范围内遵循比尔定律,加入表面活性剂TrittonX-100形成四元超分子复合物,灵敏度提高27%。和农业部颁布的NY/T1678—2008乳品测定-双缩脲法相比,灵敏度提高18倍。建立的四元反应体系光度法,操作简便,再现性好,线性范围宽,基本无干扰,用于乳品中总蛋白含量测定,和经典方法考马斯亮蓝法结果吻合。
In pH3.94 HAc-NaAc buffer medium, the azoarsenazoles reacted with Cu2 + to form a stable purplish red complex, which reacted with the biological macromolecule protein to form a complex. The λmax646nm was 541nm red (BSA). The protein follows the Beer’s law in the range of 20-100 μg / mL. The addition of surfactant TrittonX-100 to form a quaternary supramolecular complex increases the sensitivity by 27% . And the Ministry of Agriculture promulgated the NY / T1678-2008 dairy determination - biuret method compared to 18 times the sensitivity. The established quadruple reaction system photometry, simple, reproducible, wide linear range, the basic non-interference, for the determination of total protein in dairy products, and the classic method Coomassie brilliant blue method consistent.