偶氮胂羧-Ce(Ⅲ)-HSA-OP的结合反应及其光度分析应用

Carboxymethyl-Ce(Ⅲ)-HSA-OP binding reaction and spectrophotometric application

偶氮胂羧在pH3.85的HAc-NaAc缓冲介质中与Ce3+反应生成稳定的蓝紫色配合物,该配合物与生物大分子蛋白质反应形成蓝色复合物,λmax=702nm,比试剂本身的λmax=543 nm红移159 nm,ε=3.74×105L.mol-1.cm-1(HSA),蛋白质在0～90μg/mL范围内遵循比尔定律,加入表面活性剂OP形成多元复合物,灵敏度提高18%。与目前临床检验中使用的双缩脲法相比,灵敏度高17倍。方法已用于测定实际人血清总蛋白,结果满意。

In HAc-NaAc buffer at pH 3.85,Carboxymethyl could bind with Ce（Ⅲ） to form a blue-violet complex,and the complex could combine with protein to form a blue complex,which presents a maximum absor-ption at 702 nm with a red shift of 159 nm compared to Carboxymethyl itself.Its apparent molar absor ptivity and linear range were 3.76×105 L·mol-1·cm-1 and 0 ～ 90 μg/mL（HSA）.The sensitivity increased 17% in the presence of OP.The sensitivity of the method is 17 times higher than that by the Biuret method.The method is simple,fast,sensitivie,and reproducible with wide linear range.The method has been used for the determination of total protein in Human serum protein with satisfactory results.