【作者】 彭忠利；

【导师】 蒋治良；

【作者基本信息】 广西师范大学 ， 有机化学， 2001， 硕士

【摘要】 第一部分：在生理pH(7.43)条件下，人血清白蛋白(human serum albumin，简称HSA)和色氨酸(tryptophane,简称Trp)在344nm和360nm分别产生荧光峰；在688nm和720nm分别产生1／2分频荧光峰。考察了分频荧光峰与荧光峰的半峰宽之比和峰高之比，发现它们分别接近一常数。两者具有相似的荧光特性。加入在色氨酸荧光峰波长处有强吸收的物质，荧光峰和分频荧光峰强度均同时减弱；当入射光波长大于荧光峰波长时，荧光峰和分频荧光峰均不出现。依据非线性光学原理和分频原理，探讨了半峰宽与中心波长(荧光峰波长)的关系及分频荧光峰产生的原因。 第二部分：在生理pH(7.43)条件下，DNA在470nm和350nm处分别产生最强共振散射峰和次强共振散射峰。固定激发波长，扫描发射波长，DNA溶液将产生瑞利共振散射峰、分频共振散射峰和倍频共振散射峰。研究了pH、离子强度、表面活性剂等对共振散射光强度的影响，由此可推测DNA分子结构与构象的变化。依据量子电动力学原理，探讨了由普通单色光引起的各种共振散射峰产生的机理，分析了各种共振散射峰强度不同的原因。 第三部分：利用金属配阴离子与蛋白质结合成超分子离子缔合物及界面的形成，采用共振散射技术建立蛋白质定量测定方法。在2.0×10’3mol.dm’3HCl介质中，蛋白质与[Fe(CN)‘]·通过静电引力为主的分子间作用力结合成超分子离子缔合物，在350nm处产生最强共振散射光信号。基于这一现象可以测定低至0.3μg.ml’~的人血清白蛋白。工作曲线在0-12μg.ml’’范围内呈线性关系。这一新方法的灵敏度比Coomassie 广西师范大学硕士论文：HSA的分频荧光及洲A和HSA的共振散射光谱研究 亮蓝法高 3倍．研究T pH、离子强度和厂e（CN）61s－浓度对反应体系 的作用。将这一方法用子合成样品的微量蛋白质测定，结果满意．更多还原

【Abstract】 Part l:At physiological pH ( 7.43 ) human serum albumin(HSA) and tryptophane(Trp) exhibit their fluorescence peaks at 344nm and 360nm,respectively, while their 1/2 fraction frequency fluorescence peaks at 688nm and 720nm. The ratio of half peak width and the ratio of peak height (or fluorescence intensity) between 1/2 fraction frequency fluorescence peak and the fluorescence peak approach a constant respectively. The two fluorescence peaks have similar fluorescence feature and their intensities weaken at the same time while strong absorbing substance in the wavelength of Trp (or HSA) fluorescence peak is added to; the two fluorescence peaks don’t appear if the excitation wavelength is longer than that of fluorescence peak . According to the principle of nonlinear optics and fraction frequency, the relationship between half peak width and center wavelength (fluorescence peak wavelength) and the cause of fraction frequency fluorescence peak have been considered.Part II: At physiological pH(7.43),DNA exhibits the strongest and the second strong resonance scattering peaks at 470nm and 350nm,respectively. Rayleighx fraction frequency and double resonance frequency scattering peaks were found in the emission spectrum of DNA. Influence of pH?ionicstrength and surfactant on the intensity of light-scattering was studied. By this way, the variation of structure or conformation of DNA molecule has been inferred. According to the principle of quantum electrodynamics,the mechanism of all kinds of resonance scattering peaks caused by ordinary monochromatic light wasdiscussed systematically and the intensity of various resonance scattering light was also analysed.PartIII:Based on the formation of supramolecular ion-association complexes resulting from the combination of metal complex anions with proteins and the interface formation, a new method for the determination of human serum albumin (HSA) by using the technique of resonance scattering has been founded. In the medium of 2. 0 ICT’mol. dm’3 HC1, proteins may combine with [Fe(CN)6]3~ by intermolecular forces(mainly by electrostatic force) to supramolecular ion-association complexes, causing a light signal of the strongest resonance scattering at 351nm. Based on this, no less 0.3 lAg-ml"1 HSA can be determined . Its calibration curve is near over the range of Q-12\ig.mTl .The sensitivity of this method is three-fold higher than that of Coomassie brilliant blue protein assay. Influence of experimental conditions such as pH, ionic strength and concentration of [Fe(CN)6] * on the intensity of resonance scattering light was studied. Effects of surfactants (cationic, anionic and nonionic) > ami no acids and metal ions on the combination of [Fe(CN)6J3" with protein were also investigated .This method has been used for the determination of synthetic samples with satisfactory results .更多还原