以钙色素线性扫描极谱法测定血清白蛋白的研究及分析应用

Linear sweep polarographic determination of protein with calcion and its analytical appliction

在pH4.0的BrittonRobinson(B-R)缓冲溶液中钙色素与蛋白质能够发生相互作用,生成一种非电活性的超分子复合物,使钙色素在溶液中的游离浓度降低,从而造成钙色素于-0.31V处产生的二阶导数极谱还原峰电流下降而峰电位不变。在优化的结合反应条件和电化学测定条件下,峰电流的下降值同人血清白蛋白(HSA)的浓度在4.0～70.0mg/L范围内呈线性关系,其线性回归方程为ΔIp″/nA=69.93c(mg/L)-192.90,(γ=0.992)。将该方法应用于实际人血清样品的测定,结果与经典的考马斯亮蓝G250光度法一致,回收率令人满意。此方法还可应用于牛血红蛋白、卵清白蛋白等蛋白质的测定。

In pH 4.0 Britton-Robinson(B-R) buffer solution calcion(CAL) could interact with protein to form a supermolecular complex.CAL had a sensitive reductive peak at -0.31V(vs.SCE) on the hanging mercury drop electrode.After adding human serum albumin(HSA) to the above solution,the reduction peak current of CAL decreased linearly with HSA concentration from 4.0mg/L to 70.0mg/L with the regression equation ΔI_(p″)(nA)=69.93c(mg/L) -192.90,γ=0.992.This method was further applied to determine the content of HSA in blood samples and the results were in good agreement with the traditional coomassie brilliant blue G-250 spectrophotometric method.This method could also be used to determine bovine hemoglobin,egg albumin etc.with satisfactory results.