Fluorospectrometric Study on the Interaction of the Condensate of o-Vanillin and Arginine Schiff Base with Trypsin and Its Analytical Application
摘 要
应用紫外分光光度法和荧光光度法研究了邻香草醛缩精氨酸席夫碱与胰蛋白酶相互作用.试验数据采用Stern-Volmer方程和Lineweaver-Burk方程处理,证实了邻香草醛缩精氨酸席夫碱对胰蛋白酶的荧光淬灭属于静态猝灭,测得结合常数(KA)为2.5×103L·mol-1,结合位点数为1.在激发波长292 nm、发射波长339 nm处,反应体系的ΔF(即F0与F值之差)与胰蛋白酶的质量浓度在0.016 7~0.150 6 g·L-1之间呈线性关系,检出限(3σ/k)为5.02×10-3g·L-1.方法用于合成样品中胰蛋白酶的测定,回收率在94.9%~103.7%之间,相对标准偏差(n=7)为1.92%.
Abstract
UV-spectrophotometry and fluorospectrophotometry were applied to the study on the interaction of condensate of o-vanillin and arginine schiff base with trypsin.It was found that the fluorescence of trypsin was quenched due to its interaction with the condensate,and it was proved by the Stern-Volmer equation and Lineweaver-Burk equation that the fluorescence quenching was a static process.Equilibrium constant of combination (KA) and number of combination sites (n) were determined and the values obtained were 2.5×103L·mol-1 and 1 respectively.Linear relationship was found between values of ΔF(F0-F)determined at wavelengths of 292 nm (λex) and 339 nm (λem) and concentration of trypsin in the range of 0.016 7-0.150 6 g·L-1,and detection limit (3σ/k) of the method found was 5.02×10-3g·L-1.The proposed method was used in the determination of trypsin in simulated sample,giving values of recovery ranged from 94.9%-103.7%,and of RSD (n=7) of 1.92%.
中图分类号 O657.32
所属栏目 工作简报
基金项目 广西自然科学基金项目(桂科基0731015)
收稿日期 2009/8/31
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备注陶慧林(1961-),女,广西贺州人,副教授,主要从事分析化学教学与科研工作.
引用该论文: TAO Hui-lin,LIU Zheng,WANG Song-mei,GAO Jiong-yang. Fluorospectrometric Study on the Interaction of the Condensate of o-Vanillin and Arginine Schiff Base with Trypsin and Its Analytical Application[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2010, 46(11): 1269~1272
陶慧林,刘峥,王松梅,高炅杨. 荧光光谱法对邻香草醛缩精氨酸席夫碱与胰蛋白酶相互作用的研究及其应用[J]. 理化检验-化学分册, 2010, 46(11): 1269~1272
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【6】冯小珍,夏金虹,刘峥.氨基酸席夫碱配合物的制备及性能研究进展[J].化学研究与应用,2007,19(3):238-243.
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