酸性铬蓝K与卵白蛋白相互作用的分光光度研究及应用

Spectrophotometric Study of Interaction between Ovalbumin with Acid Chrome Blue K and Its Application

在pH1.30的氯乙酸缓冲溶液中,在TritonX-100存在下,卵白蛋白与酸性铬蓝K可形成有色复合物,其最大吸收波长为585nm。卵白蛋白浓度在0～2.85mg/L和3.14～11.40mg/L范围呈线性,摩尔吸光系数分别为4.76×105L·mol-1·cm-1、5.46×104L·mol-1·cm-1。相对标准偏差在1.18%～1.27%,回收率在98.8%～101.7%之间。提出的方法已用于豆粕浸提液中蛋白质的测定,分析结果满意。探讨了卵白蛋白与酸性铬蓝K形成复合物的反应机理。

In pH 1.30 chloroacetate buffer solution and in the presence of Triton X100, the ovalbumin(OVA) reacts with acid chrome blue K(AK) to form a colored complex. The maximum absorption of the complex is at 585 nm. The regression equations and molar absorptivities of the two linear ranges of 0~2.85 mg·L-1 and 3.14~11.40 mg·L-1 for OVA are A=0.0070+0.1074 c(mg·L-1) with ε=4.76×105 L·mol-1·cm-1 and A=0.0610+0.0799 c(mg·L-1) with ε=5.46×104 L·mol-1·cm-1 respectively. The proposed method has been used for the determination of proteins in extracted solutions of soya residue with satisfactory results. The relative standard deviations and recoveries are 1.18%~1.27% and 98.8%~101.7%, respectively. The reaction mechanism of OVA with AK is briefly discussed.