【作者】 高静；

【导师】 刘保生；

【作者基本信息】 河北大学 ， 分析化学， 2005， 硕士

【摘要】 荧光共振能量转移是一种通过荧光物质间发生荧光能量转移进行分析的实验方法。由于其具有比光度法灵敏度高,与常规荧光法和共振光散射相比受瑞利散射光干扰小,重现性好的特点,该方法已被广泛的应用于化学分析、生物结构研究等领域。本文就此类方法进行了进一步应用研究,共分五章: 第一章文献综述,介绍了荧光共振能量转移方法的基本原理、发展及其在各方面的应用。第二章研究了吖啶橙(AO)-罗丹明6G(R6G)荧光共振能量转移光谱,考察了体系的光谱特征和主要影响因素,建立了人血清白蛋白(HSA)、牛血清白蛋白(BSA)、卵清白蛋白(OVA)等的测定新方法。在pH = 7.20 的Tirs-HCl 缓冲溶液,十二烷基苯磺酸钠介质中,AO–R6G 能够发生有效能量转移,蛋白质的加入使AO–R6G 体系的荧光猝灭。在优化的实验条件下,牛血清白蛋白(BSA)、人血清白蛋白(HSA)工作曲线线性范围分别为:0~31.25、0~30.00 mg·L-1; 检出限分别为:0.32、0.33 mg·L-1;平行六次测定相对标准偏差为1.07~1.97 %;回收率为96.7~103.2 %,并将该方法用于人血清样品中蛋白质的测定,结果与医院常用的双缩脲法一致。第三章建立了应用吖啶橙(AO)-罗丹明6G(R6G)能量转移体系测定维生素B12的方法。在pH = 5.0 的Na2HPO4-柠檬酸缓冲溶液,十二烷基苯磺酸钠介质中,AO-R6G间发生能量转移,而维生素B12(VB12)的加入可使AO–R6G 体系的荧光猝灭。维生素B12 工作曲线的线性范围为:0~3.0×10-5 mol·L-1; 检出限为:4.8×10-7 mol·L-1;平行六次测定相对标准偏差为0.51~0.64 %;回收率为98.40~103.62 %。应用于维生素B12注射液中维生素B12含量的测定。第四章研究了利用吖啶橙(AO)-罗丹明B(RB)共振能量转移荧光猝灭法测定痕量砷的方法。在λex /λem = 470/580 nm,0.016 mol·L-1 H2SO4溶液中,十二烷基苯磺酸钠(DBS)存在下,砷与钼酸铵生成砷钼杂多酸使能量转移体系罗丹明B 的荧光猝灭。砷在0.01~0.25 mg·L -1范围内与罗丹明B 荧光猝灭程度呈良好的线形关系,方法检出限为2.56 μg·L -1。该方法可应用于茶叶中痕量砷的测定,结果满意。第五章建立了弹性共振散射测定硫酸庆大霉素的新方法。在pH= 5.29 的KH2PO4-更多还原

【Abstract】 Fluorescence resonance energy transfer is an analytical method by transfering energy between fluorescent substances. The method has higher sensitivity than the luminosity method .Compared with fluorescence method and resonance light scattering method , the method is characterized by few inferring substances, good reproduction .It is applied to the chemical analysis , the biological structure studying . This paper has carried on the study by this kind of method . This paper consists of five chapters: Chapter one: It is an introduce of the basic theory and applications of fluorescence resonance energy transfer techniques. Chapter two: Based on the AO-R6G energy transfer systems a novel fluorescence quenching method for the determination of proteins has been presented. It was found that the effective energy transfer could occur between AO and R6G in the dodecyl benzene sodium sulfonate solution, Tris-HCl buffer solution at pH=7.20. The fluorescence intensity of AO-R6G systems was diminished by proteins in alkalescence medium . Under optimal conditions, the linear ranges of calibration curves for the determination of bovine serum albumin （BSA）and human serum albumin （HSA） were 0³1.25 mg·L^-1 and 0³0.00 mg·L^-1 respectively. Among six times determination , the relative standard deviation was 1.07¹.97 % and the recovery was 96.7¹03.2 % . The detection limits were 0.32 mg·L^-1 for BSA , and 0.33 mg·L^-1 for HSA. The determination results of the proteins in human serum samples by this method are very close to those obtained using biuret spectrophotometric method. Chapter three: Based on the AO-R6G energy transfer systems a novel fluorescence quenching method for the determination of Vitamin B12 has been presented. It was found that the effective energy transfer could occur between AO and R6G in the dodecyl benzene sodium sulfonate solution, Na2HPO4-citric acid buffer solution at pH = 5.0. The fluorescence intensity of AO-R6G systems was diminished by Vitamin B12 in alkalescence medium . The linear range of calibration curves for the determination of Vitamin B12 was 0 3.0×10-5 mol·L^-1. The detection limits were 4.8×10-7 mol·L^-1 for Vitamin B12 .Among six times determination , the relative standard deviation was 0.51 0.64 % and the recovery was 98.40¹03.62 % . It can be satisfactorily used in the determination of the injection content of更多还原