同步荧光光谱法测定蛋白质

Synchronous Fluorimetric Determination of Protein

在模拟生理条件下，由于核苷类药物中间体氰基乙基尿嘧啶（CEU）与血清白蛋白相互作用，血清白蛋白的内源荧光发生特异性变化，且体系的同步荧光强度和溶液中血清白蛋白的浓度呈线性关系，据此提出以氰基乙基尿嘧啶为探针，用固定波长同步荧光光谱法测定人血清白蛋白（HsA）和牛血清白蛋白（BSA）的方法。在最佳试验条件下，体系的荧光强度与HSA和BSA的质量浓度分别在1．38-496．2mg·L^-1和1．56-624．0mg·L^-1范围内呈线性关系，检出限（3S／N）分别为0．045mg·L^-1和0．051mg·L^-1。方法应用于人血清及牛血清中HSA及BSA的测定，并以此样品为基体分别加入HSA及BSA标准溶液作回收试验，测得回收率在95．1％～102．5％之间，相对标准偏差（n=6）在0．43％～2.72％之间。

Under the simulated physiological conditions, by the interaction of serum albumin and cyanoethyl uracil （CEU）, an intermediate of nucleoside drugs, an extraordinary change of inherent fluorescence of serum albumin was observed; and linear relationship between the magnitude of synchronous fluorescence intensity and concentration of serum albumin in the solution was obtained. Based on these facts, a new method of synchronous fluorimetric determination of human serum albumin （HSA） and bovine serum albumin （BSA）, using CEU as probe was proposed. Ranges of linearity for HSA and BSA were found to be 1.38-496.2 mg · L^-1 and 1.56-624. 0 mg · L^-1 , with values of detection limit （3S） 0. 045 mg · L^-1 and 0. 051 mg· L^-1 , respectively. The proposed method was applied to the determination of HSA in human serum sample and BSA in bovine serum sample, and on the base of these samples, standard solutions of HSA and BSA were added to test for recovery of the method. Values of recovery and RSD＇ s （n = 6） found were in the ranges of 95. 1% --102. 5% and 0. 43% - 2. 72% respectively.