【作者】 齐剑英；

【导师】 冯德雄； 杨培慧；

【作者基本信息】 暨南大学 ， 分析化学， 2003， 硕士

【摘要】 谷胱甘肽是生物体内重要的非蛋白巯基三肽，在生命科学、食品、环境保护以及医药卫生等领域都有非常广泛的应用。故建立高选择性、高灵敏度简便快速的谷胱甘肽的分析测定方法具有重要意义。 本文研究了还原型谷胱甘肽（GSH）与不同荧光增强剂的荧光、紫外等光学性质，并建立了GSH新的荧光检测方法。同时本文进行了金属离子对GSH-OPA体系荧光的影响的实验。一些有重要意义的结果如下： 1 在pH8.0的磷酸缓冲溶液中，OPA对GSH有较好的荧光增强作用，并应用OPA作为GSH的荧光增强剂，建立了GSH的荧光检测方法，GSH的浓度线性范围为2.8×10^-8～9.8×10^-6mol/L，测定限为2.8×10^-8mol/L。相对标准偏差为4.3％。 2 研究了镧系离子和Zn²⁺、pb²⁺、Cu²⁺等离子对GSH-OPA体系荧光强度的影响。实验中发现仅Zn²⁺能够对GSH-OPA体系荧光有增敏作用。同时Zn²⁺还能够提高GSH-OPA体系的稳定性，据此建立了加入Zn²⁺作为荧光增强剂测定GSH的检测方法，可降低GSH-OPA的荧光检测法的检测限。 3 研究发现两种GSH的新有机荧光增强剂（4-氨磺基-7氟-2,1,3苯并二唑；荧光胺），并建立了相应GSH的新荧光检测方法。浓度线性范围分别为5.6×10^-8～8.4×10^-6mol/L和4.2×10^-7～9.8×10^-6mol/L。 4 应用建立了的GSH荧光检测方法测定了实际样品中GSH的含量，并与常用的HPLC方法进行了对比，结果满意。所建立的方法简便快速，更多还原

【Abstract】 Glutathione , a tripeptide with one sulfhydryl group ,is present in many physiological fluid , animal and microbial cells. It plays important biologic function in the organism ,including protein and DNA synthesis ,enzyme activity, metabolism and cell protection. And It has very wide applications in the life sciences, food, environment protection , physic field and so on. Essential to ongoing investigations of this tripeptide are assay techniques which are sensitive, specific, and easily perform with readily available materials.The optics character of GSH had been studied in this work. The new detect methods of GSH had been established. And the effect of metal ions on fluorescence intensity of GSH-OPA was studied too. The result is as follow.1 The fluorescence system of GSH-OPA has good fluorescence signal with phosphoric acid buffer (pH8.0) . And GSH could be determined using OPA with linear range of 2.8 × 10-8-9.8× 10-6mol/L. The detect limit is 2.8 × 10-8 mol/L, presenting relative standard deviation 4.3 percent.2 The effect of lanthanum ions ,zinc ion, plumbum ion, copper ion on fluorescence intensity of GSH-OPA had been studied. And the result shows that only zinc ion can enhance the fluorescence intensity of GSH-OPA. Simultaneity, the zinc ion also can improve stability . On the basis of zinc ion can improve fluorescence intensity of GSH-OPA, the determination of GSH had been established, with low detect limit.3 Two new fluorescence intensifiers of GSH had been found, 4-aminosulfonly-7-fluoro-2,l,3-benzoxidiazole and Fluram. Then using the two intersifiers,GSH could be determined with the linear range of 5.6-10-8-8.4× 10-6mol/L and 4.2 × 10-7-9.8× 10-6mol/L respectively, with determination limit 5.6 -10-8 mol/L and 4.2 ×10-7 mol/L.4 The new methods were applied to detect GSH in samples . Moreover, the methods were contrasted with HPLC, with satisfactory results. with easily operation and low determination limit.更多还原