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中文摘要：
研究了邻羟基苯基荧光酮（o-HPF）荧光光度法测定牛血清白蛋白（BSA）的反应条件,在pH 7.90的B-R缓冲溶液中,o-HPF与BSA作用形成稳定的结合物,导致体系荧光猝灭,依据其荧光猝灭程度与外加BSA量的关系建立了定量测定BSA的方法。该方法具有良好的选择性和稳定性,实验操作简单,干扰少,灵敏度较高,测定BSA线性范围为1.32～18.54μg·mL^-1,回归方程为ΔF=431.51c（10-7mol·L^-1）＋457.78,相关系数r=0.997。测定方法中重金属离子如Pb（Ⅱ）等对BSA测定允许量较低,其他物质的存在对BSA的测定干扰较小。通过对o-HPF与BSA作用荧光猝灭常数的测定及热力学常数计算,讨论了o-HPF与BSA的作用机理,表明o-HPF与BSA作用方式主要为静电引力的非共价作用,BSA对o-HPF荧光猝灭为分子间形成了结合物而引起的静态猝灭。
英文摘要：
The reaction conditions for the determination of bovine serum albumin （BSA） using o-hydroxyphenylfluorone （o- HPF） as fluorescence probe reagent were studied. In the B-R medium at pH 7. 90, o-HPF could react with BSA, producing a stable complex compound which resulted in fluorescent quenching of this binary system. The quantitative assay was carried out based on the relation between the degree of fluorescent quenching and the additional amount of BSA. The method has advantages of good selectivity, stability and simplicity. The linear range of o-HPF fluorescence spectrophotometric method was 1.32-18. 54 μg - mL-1. Moreover, the mechanism of the reaction BSA and o-HPF was discussed, and the quenching constant, and the thermodynamics constant of the fluorescence effects were calculated. The results showed that the nonconvalent binding forces were the binding force between o-HPF and BSA, and the quenching of BSA to o-HPF was probably a single static quenching process.
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