【作者】 王明瑞；

【导师】 董学芝；

【作者基本信息】 河南大学 ， 分析化学， 2008， 硕士

【摘要】 近年来,随着我国畜牧业的不断发展,畜禽品种和数量有了很大幅度的增加。相应的,我国畜禽疾病的种类和发生频率也大大增加,这给我国的畜禽业造成了巨大的经济损失。因此,畜禽疾病的快速诊断是很重要的研究课题。血清白蛋白是血液中含量最丰富的蛋白质,它在动物体液中的含量变化可以作为诊断疾病的依据。利用新研究的光谱探针检测动物体液中蛋白质的含量,并应用于动物疾病的诊断和防治,是一项非常有意义的工作。本文重点开展了有机小分子二苯胺磺酸钠,二甲酚橙及萘酚绿B等光谱探针与蛋白质的相互作用研究,并将研究应用于动物体液中蛋白质含量的测定。论文主要内容如下:一、综述了光谱分析法中分光光度法、荧光光度法、共振散射光度法在蛋白质定量分析中的研究现状。二、研究了二甲酚橙与Cu(II)配合物与牛血清白蛋白的共振散射光谱的特征,考察了体系主要影响因素;然后建立了牛血清白蛋白(BSA)的测定新方法,并将方法用于牛血清样品中蛋白质的测定,结果与文献方法一致。三、研究了二苯胺磺酸钠探针与牛血清白蛋白的共振散射光谱的特征,考察了体系主要影响因素;然后建立了牛血清白蛋白(BSA)的测定新方法,并将方法用于牛血清样品中蛋白质的测定,结果与文献方法一致。四、应用紫外可见光谱、荧光光谱法研究了萘酚绿B与牛血清白蛋(BSA)之间的相互作用。实验表明:在水溶液体系中,萘酚绿B对血清白蛋白的荧光有较强的猝灭作用,其荧光猝灭主要为静态猝灭。进一步从荧光猝灭数据求得不同温度下染料与BSA的结合常数K,发现随反应温度上升K值下降。由反应焓变、熵变,确定萘酚绿B与血清白蛋白的结合疏水力作用为主。利用同步荧光光谱和UV/VIS吸收光谱,分析了NGB的加入对BSA的构象的影响。五、应用紫外可见光谱研究了萘酚绿B与牛血清白蛋(BSA)之间的相互作用。实验表明:在高温的酸性溶液中,萘酚绿B发生褪色反应;牛血清白蛋的加入对萘酚绿B的褪色反应有阻抑作用,由此进一步确定萘酚绿B与牛血清白蛋在水溶液中可以发生结合,形成稳定的复合物。更多还原

【Abstract】 In recent years, along with our country animal husbandry’s unceasing development, the poultry variety and quantity had great scope increase. Corresponding, our country’s livestock and poultry diseases and the type of frequency also greatly to increase, significantly, this has created the huge economic loss for our country’s poultry industry. Therefore, the rapid diagnosis of animal diseases is very important research subject.The serum albumin is the content richest protein in the blood, which in animal body fluids in the content of a certain extent, can be used as the basis for the diagnosis of the disease. Using the new research discovery’s spectrum probe examination protein’s content, and applied animal protein in the body fluids or the determination of biological products in the determination of protein, is a very meaningful work..This article focuses on small organic molecules diphenylamine sulfonic acid sodium, the XO and naphthol Green B and protein interactions, applied research and animal body fluids in the determination of protein.Its paper primary coverage is as follows:The results and methods for study of reaction mechanism between small organic molecules with protein were reviewed with 84 references.The xylenol orange and the Cu(II) complex probe to determine the protein content have been studied. The Resonance light scattering spectrum of the characteristics of the XO and Cu (II) complex with bovine serum albumin have been studied. This method was applied to the determination of bovine urine and compared to Bradford method with satisfactory results.The diphenylamine sulfonic acid sodium probe to determine the protein content have been studied by resonance scattering spectrum (RLS). The diphenylamine sulfonic acid sodium with bovine serum albumin resonance scattering spectrum of the characteristics have been studied. This method was applied to the determination of bovine urine and compared to Bradford method with satisfactory results.The binding characteristics of Naphthol green B(NGB) with bovine serum albumin(BSA) have been studied by spectroscopy method in aqueous solution. It is proved that static quenching exits between NGB-BSA super-molecular complex. The formation constant KA and the thermodynamic functions (such as△G、△H and△S) for the reaction have all been measured; temperatures on the formation constant of Naphthol green B with BSA was also studied; According to the thermodynamic parameters, the hydrogen bonding and van’der Waals forces played major role in the interaction. The effect of Naphthol green B on the conformation of BSA has also been analyzed using synchronous fluorescence spectroscopy; The binding distance between Naphthol green B and BSA and the transfer efficiency have been obtained based on the mechanism of Forster energy transfer.Studies discovered: In the high temperature acid solution, naphthol green B has the discoloration response; BSA’s accession to the naphthol Green B inhibition reaction to the fading effect, which further define naphthol Green B and bovine serum albumin can bind.更多还原