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  

Fluorospectrometric Study on the Interaction of the Condensate of o-Vanillin and Arginine Schiff Base with Trypsin and Its Analytical Application

作　　者：陶慧林[1] 刘峥[1] 王松梅[1] 高炅杨[1] 

机构地区：[1]桂林理工大学化学与生物工程学院,桂林541004

出　　处：《理化检验（化学分册）》2010年第11期1269-1272,共4页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：广西自然科学基金项目(桂科基0731015)

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-Vol mer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%。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%.

关 键 词：荧光光谱法 胰蛋白酶 邻香草醛缩精氨酸席夫碱 相互作用 

分 类 号：O657.32[理学—分析化学]