蛋白质的光分析化学特性研究

【作者】 杨平华；

【导师】 张正奇；

【作者基本信息】 湖南大学 ， 分析化学， 2002， 硕士

【摘要】 本文在查阅了大量的利用染料测定蛋白质及染料与蛋白质作用的研究论文的基础上，较系统的研究了偶氮胂Ⅰ、7-苯基偶氮-1，8-二羟基萘-3，6-二磺酸钠、偶氮氯膦Ⅰ和偶氮氯膦Ⅲ等测定蛋白质的方法。同时，也研究了偶氮胂Ⅰ、7-苯基偶氮-1，8-二羟基萘-3，6-二磺酸钠和偶氮氯膦Ⅰ等与蛋白质作用的机理。 1．研究了蛋白质与偶氮胂Ⅰ的显色反应。在pH3.2的Britton-Robinson缓冲溶液中，偶氮胂Ⅰ与蛋白质形成红色复合物，λ_max为551nm，ε为2.5×10⁵L.mol^-1.cm^-1。蛋白质含量在13.2mg.L^-1-231mg.L^-1范围内服从Beer定律。所提出的方法可直接用于血清、花生等生物样品中蛋白质含量的测定。 2．研究了蛋白质与7-苯基偶氮-1，8-二羟基萘-3，6-二磺酸钠的显色反应。在pH2.7的Britton-Robinson缓冲溶液中，试剂与蛋白质形成红色复合物，λ_max为564nm，人血清白蛋白含量在6.6mg.L^-1-66mg.L^-1范围内服从Beer定律。所提出的方法可直接用于血清、花生等生物样品中蛋白质含量的测定。 3．研究了蛋白质与偶氮氯膦Ⅲ的显色反应。在pH2.0的Britton-Robinson缓冲溶液中，偶氮氯膦Ⅲ与蛋白质形成蓝色复合物，复合物的λ_max为685nm。其表观摩尔吸光系数ε=1.8×10⁵L.mol^-1.cm^-1。吸光度与蛋白质含量呈线性关系，校正曲线在6.6mg.L^-1-79.2mg.L^-1间服从Beer定律。应用本法可直接测定血清、花生等生物样品中蛋白质含量。 4．研究了偶氮氯膦Ⅰ与蛋白质复合物的光度性质。在pH2.30的Britton-Robinson缓冲溶液中，偶氮氯膦Ⅰ与蛋白质形成红色复合物，λ_max为562nm，蛋白质含量在小于52.8mg.L^-1时服从Beer定律。 5．应用分光光度法研究了蛋白质与偶氮胂Ⅰ、7-苯基偶氮-1，8-二羟基萘-3，6-二磺酸钠和偶氮氯膦Ⅰ等的相互作用。用摩尔比法测定了它们的最大结合数，研究了它们的结合机理，并进一步讨论了离子强度对它们结合的影响。更多还原

【Abstract】 After consulted more than 100 articles about dyes interact with proteins. In this paper,Arsenazo I,7-Phenyl azo-1,8-Dihydroxyl naphthaiene-3,6-disulfo sodium,Chlorophosphonazo III and Chlorophosphonazo I being used to quantity analysis of proteins has been studied. In the same time,the mechanism of their combination has been studied too.1. In a pH 3. 2 Britton-Robinson buffer solution Bovine Serum Albumin (BSA) forms a red complex with Arsenazo I . Its maximum absorption peak is 551nm with molar absorption of 2. 5X 10L. mol"1. cnf’. Beer’ s law is obeyed in a range from 13. 2mg L-’ to 231mg L"’.The proposed method has been applied to determination of protein in serum and peanut samples2. In a pH2. 7 Britton-Robinson buffer solution Human Serum Albumin (HSA) forms a red complex with 7-Phenyl azo-1,8-Dihydroxyl naphthaiene-3,6-disulfo sodium .Its maximum absorption peak is 564nm. Beer’ s law is obeyed in a range from 6. 6mg. I/1 to 66mg. I/’. The proposed method has been applied to determination of protein in serum and peanut samples.3. In a pH2. 0 Britton-Robinson buffer solution bovine Serum Albumin forms a blue complex with Chlorophosphonazo III. Its maximum absorption peak is 685nm. Beer’ s law is obeyed in a range from 6. 6mg I/1 to 79. 2mg I/1. The proposed method has been applied to determination of protein in serum and peanut samples.4. In a pH2.3 Britton-Robinson buffer solution Bovine Serum Albumin forms a red complex with Chlorophosphonazo I . Its maximum absorption peak is 562 nm. Beer’ s law is obeyed in less than 52. 8mg I/1.5. The interaction of Arsenazo I,7-Phenyl azo-1,8-Dihydroxyl naphthaiene-3,6-disulfo sodium,and Chlorophosphonazo I with bovine Serum Albumin were studied. The principle of molar ratio method was applied to determine the maximum combination number of dye bound on proteins. The mechanism of their combination has been studied. It is found that sodiumchloride concentration of the solution has effect on their combination.更多还原