首页> 中文期刊>光谱学与光谱分析 >荧光法研究琥珀酸曲格列汀与牛血清白蛋白的相互作用及分析应用

荧光法研究琥珀酸曲格列汀与牛血清白蛋白的相互作用及分析应用

     

摘要

琥珀酸曲格列汀(QGLT)是一种新型降糖药,基于其能够猝灭牛血清白蛋白(BSA)的荧光,用荧光法研究了两者的相互作用.对影响荧光猝灭的因素如溶液酸度、介质性质等进行了优化.猝灭机理探讨发现QGLT主要通过静态猝灭降低BSA的荧光,并推导出两者的结合位点数为n=1,结合常数分别为4.529×104 L·mol-1(298k)和2.958×104 L·mol-1(303 k).热力学参数测定结果显示吉布斯自由能变、焓变和熵变均为负值,表明QGLT和BSA的结合主要是通过氢键与范德华力自发结合的.研究中发现Lineweaver-Burk方程具有良好的线性关系,线性方程为:(F0-F)-1=2.711×10-4+3.51×10-3 cQGLT-1(cQGLT:μg·mL-1),r=0.9989,检出限为0.13μg·mL-1(S/N=3),线性范围为0.5~10.0μg·mL-1.回收率实验结果表明方法的准确性较好(回收率为94.0%~97.5%).用该方程测定了药片和人血清中QGLT含量,结果令人满意.本研究拓展了QGLT含量测定的新方法.%In this paper the interaction between trelagliptin succinate (QGLT) and bovine serum albumin (BSA) was studied by fluorescence spectrometry .The factors affecting fluorescence quenching such as solution acidity and medium characteristics were optimized .The examination results of quenching mechanism showed that the fluorescence intensity of BSA was quenched strongly by QGLT mainly through a static quenching procedure ,the number of binding sites was 1 ,and the biding constants was 4.529 × 104 L · mol-1 (298 k) or 2.958 × 104 L · mol-1 (303 k) ,respectively .The determination results of thermodynamic parameters show that the change of Gibbs free energy ,enthalpy or entropy was negative ,which clarified there was an automatic reaction happened between QGLT and BSA ,and both hydrogen bonds and van der Waals force played a role in the binding of QGLT to BSA .It was also found in the examination that Lineweaver-Burk equation [(F0 - F)-1 =2.711 × 10-4 + 3.51 × 10-3 cQGLT -1 (cQGLT :0.5~10.0 μg · mL -1 )] owned better linear relationship (r=0.9989) with the detection limit of 0.13 μg · mL -1 .The recovery experiment results showed that the method owed good accuracy (the recovery is 94.0% ~ 97.5% ) and the proposed method has been successfully applied to determine QGLT in QGLT tablet and human serum .The examination developed a new method to determine the content of QGLT .

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