光谱法研究根皮苷与人血清白蛋白相互作用.docVIP

光谱法研究根皮与人血清白蛋白相互作用 摘 要在模拟生理条件下，采用荧光光谱紫外光谱研究了根皮Phlorizin) 与人血清白蛋白（HSA）的相互作用。：根皮293K、303K和313根皮结合常数分别为×105、1.774 8×105、1.193 5×105 L?mol-1?s-1，结合位点数n近似为1；热力学根皮与HSA间结合力为；根据F?rster非辐射能量转移理论求得二者结合距离为 nm；同步荧光光谱。 关键词根皮；人血清白蛋白；荧光；Study on Interaction between Phlorizin and Human Serum Albumin by Spectroscopic Methods Abstract: The interaction of Phlorizin with human serum albumin (HSA) was studied by fluorescence spectrometry, UV-Visible absorption spectrometry and synchronous fluorescence spectrometry. The results showed that Phlorizin could induced an endogenous fluorescence quenching of HSA under a mechanism of static quenching. The binding constants were determined to be 3.163 5×105(293K), 1.774 8×105 (303K) and 1.193 5×105(313K), respectively. The binding site (n) was about 1. The thermodynamic parameters suggested that the interaction forces between Phlorizin and HSA were mainly hydrogen and vander waals force., The binding distance was estimated to be 3.97 nm according to F?rster nonradiation energy transfer mechanism. The results synchronous fluorescence spectra of Phlorizin-HAS system showed that Phlorizin mainly interacted with tryptophan in HSA, leading to a local change in protein conformation. In addition, the addition of the common ions could affected the binding constant of Phlorizin-HSA complex. Key words: phlorizin; human serum albumin; fluorescence quenching; thermodynamic parameter 根皮Phlorizin)是一种具有明显生物活性的天然化合物，其化学成分是1-(2-(β-D-吡喃葡萄糖氧基)-4,6-二羟基苯基)-3-(4-羟基苯基)-丙酮根皮本采用荧光光谱根皮根皮根皮根皮根皮 1 实验部分 1.1仪器与试剂 -5301型荧光分光光度计（日本公司）；UV-2550紫外可见分光光度计（日本岛津公司）；501A型超级数显恒温水浴（上海浦东荣丰科学仪器有限公司）人血清白蛋白（9%，）HSA配制成1.5×10-5 mol·L-1标准液（北京索莱宝科技有限公司）成×10-5 mol·L-1标准溶液；缓冲溶液(pH 7.4)；水为二次蒸馏水，其它试剂均为分析纯。 1.2实验方法 以定容至恒温水浴30 min以285 nm为激发波长激发和发射狭缝均为5 nm△λ=15、60 nm 2 结果与讨论 2.1 对HSA的荧光猝灭光谱 HSA发生荧光猝灭。 图1对HSA的荧光猝灭光谱（K） Fig.1 Fluorescence quenching spectra ofHSA system 303 K；c HSA=1.05×10-6 mol·L-1; c 根皮苷: 0.0, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0（(10-6 mol·L-1） 2.2 荧光猝灭机理及猝灭常数的测定 引起