【作者】 潘惠英；

【导师】 陈立功；

【作者基本信息】 天津大学 ， 药物化学， 2005， 硕士

【摘要】 氨基荧光素是荧光素类染料中重要的荧光探针之一,其活性氨基基团能与生物分子中羧基相结合,结合后荧光强度大大增强,因而广泛的应用于蛋白质分析和免疫学研究。本文以4-硝基邻苯二甲酸,间苯二酚为原料在ZnCl2催化下通过缩合、还原,在6%盐酸中重结晶分离制备了5-氨基荧光素和6-氨基荧光素。该方法与传统5-氨基荧光素制备方法相比较,反应步骤缩短,5-氨基荧光素的总收率由18%提高至40%。5-氨基荧光素虽能与羧基反应,但是由于其氨基与芳基相连,大大降低其衍生化的能力,且其为水溶性荧光染料,亲脂性差,限制了其在生物分析中的应用。为改善其荧光性能,提高氨基的反应能力,我们又设计合成一系列5-氨基荧光素的衍生物。分别以5-氨基荧光素和2’,7’-二氯氨基荧光素为原料进行异硫氰酸化制备了5-异硫氰酸酯荧光素和2’,7’-二氯异硫氰酸酯荧光素;通过酰化制备了5-(2-氯乙酰胺基)荧光素以及5-丁二酰亚胺基荧光素;与三聚氯氰反应制备了5-[2-(4,6-二氯三嗪基)氨基]荧光素;所得衍生物的化学结构由IR,MS,NMR确证,并测定了衍生物的荧光性能,发现所合成的氨基荧光素衍生物与5-氨基荧光素相比其荧光强度、最大激发波长和最大吸收波长均有明显变化,显示了较优越的荧光性能。多芳环氧鎓盐本身也是无机离子显色和荧光分析的具有较高灵敏度的荧光探针。本文制备了一系列带有不同取代基团的吡喃鎓盐,使之与5-氨基荧光素反应,得到了双发色团荧光染料1-(5-荧光素)吡啶鎓盐,其中1-(5-荧光素)-2,4,6-三甲基吡啶鎓盐显示了良好的荧光性能。更多还原

【Abstract】 Aminofluorescein is one of the most important fluorescent probes. The fluorescent intensity of aminofluorescein derivatives is enhanced after the amine group is bonded to carboxyl group of the biomolecules. So aminofluorescein has been widely applied in the protein analysis and immunology.In this paper, aminofluorescein was obtained from 4-nitrophthalic acid, resoreinol in the presence of zinc chloride through condensation and reduction, and 5-aminofluorescein and 6-aminofluorescein were successfully isolated by recrystallization of the obtained mixture of 5- and 6-aminofluorescein in 6% (w) hydrochloric acid. Compared with the traditional method, this synthetic route is more convenient and the total yield of 5-aminofluorescein was increased from 18% to 40%.The ability of aminofluorescein linking with carboxyl group was reduced due to the conjugation of amine group with aromatic ring. Aminofluorescein was water-soluble fluorescent probe, which restricted its application in biological analysis. In search of new better fluorescent probes, a series of derivatives of aminofluorescent were designed and synthesized such as fluorescein isothiocyanate, 2’,7’-dichloro- fluorescein-5(6)-isothiocyanate, 5-(4,6-dichlorotriazin-2-yl)aminofluorescein, fluore- scein-5-chloroacetamide, 5-succinimidylfluorescein. The structures of the derivatives were confirmed by IR, MS and NMR, and their fluorescent properties were determined. The fluorescent intensity, maximum excitation wavenumber and maximum emission wavenumber of the derivatives were changed compared with aminofluorescein, which indicated better fluorescent properties.Besides, the compounds of 2,4,6-triarylpyrylium salts were sensitive fluorescent probes in the dyeing of inorganic ions and fluorescent analysis. Pyrylium perchlorates were prepared. The dyes with double fluorophore 1-(fluorescein-5-yl) pyridinium salts were yielded by the treatment of pyrylium salts with 5-aminofluorescein. 1-(Fluorescein-5-yl)-2,4,6-trimethylpyridinium perchlorate showed good fluorescent properties.更多还原