新型荧光试剂的合成及其在高效液相色谱中的应用研究

【作者】 张海峰；

【导师】 尤进茂；

【作者基本信息】 曲阜师范大学 ， 物理化学， 2006， 硕士

【摘要】 色谱是一种分离分析技术,自20世纪四十年代以后,逐渐得到发展,而且其发展越来越迅速。从技术到理论,再到各种分离模式,以及在生物化学、医学、药物临床、化学化工、食品卫生、环保检测、商品检验和法医检验等领域的应用,都得到了突飞猛进的发展。现在色谱已经成为分析化学学科中的一个重要分支。然而由于样品量及相应组分的低含量限制,并非所有的化合物都可直接用于色谱分析,所以提高检测灵敏度是十分必要的。为此利用化学衍生反应达到改变被测化合物特性的目的,是提高检测灵敏度的一个重要手段。 由此本文设计合成了两种高灵敏度荧光试剂并对其在高效液相色谱中的应用进行了研究,结果令人满意。 第一章:荧光原理和化学衍生化法的简介,高效液相色谱的发展和相关的基础知识的简单综述。 第二章:荧光试剂1,2-苯并-3,4-二氢咔唑-9-羟乙基肼基甲酸酯的合成,对其荧光性质进行了考察。研究了该试剂在高效液相色谱中的应用,包括三部分: 2.1 1,2-苯并-3,4-二氢咔唑-9-羟乙基肼基甲酸酯用于脂肪醛的高效液相色谱分析,对衍生条件、沈脱参数进行了优化,并应用于饮料样品中醛的分离分析。 2.2 1,2-苯并-3,4-二氢咔唑-9-羟乙基肼基甲酸酯用于中性糖的高效液相色谱分析。对衍生条件、洗脱参数进行了优化。考察了中性糖衍生物的检测限、线性范围和回归方程。 2.3 1,2-苯并-3,4-二氢咔唑-9-羟乙基肼基甲酸酯用于类固醇的高效液相色谱分析。对衍生化条件、洗脱程序等进行了优化,并将所建立的方法用于老鼠粪便中的类固醇的分析。 第三章:合成了2-[2-(对甲苯磺酸酯)乙基]哟唑并[3,4-b]-7-甲基喹啉,并用于脂肪酸的高效液相色谱分析。分析了苔藓中脂肪酸的含量,结果令人满意。更多还原

【Abstract】 Chromatography is a kind of analytical techniques and its development in theory and technology has been growing rapidly from 1940s. The enthusiastic adoption of high-performance liquid chromatographic method has had a great impact on chemical analysis, not only in chemistry and biochemistry but also in pharmacology, toxicology, clinicalscience, genetics, forensic science , environmental science, quality control in industry and many other fields. This method is one of the fastest developing and the most widely used fields in analytical chemistry. However not all compounds are accessible to direct analysis by chromatography due to the limitations of properties of the compounds themselves. Therefore derivatization of the analytes with fluorescent reagents has been widely adopted. It can alter the chemical and physical properties of target compounds to make them easier to analyze. Currently, fluorescence detection is still recognized to be one of the most sensitive detection technologies. In this paper, we designed and synthesized two novel fluorescence reagents with the different chromophores. The optimal derivatization conditions such as buffer pH, reaction time and solvent system are investigated. At the same time, fluorescence excitation, emission spectra of reagents and the suitability of the developed method for the analysis of real samples are evaluated.Chapter one: The principles and applications of fluorescence detection and fluorescence reagents in high performance liquid chromatography (HPLC) are systematically reviewed.Chapter two: The synthesis of fluorescence reagent 1,2-benzo -3,4-dihydrocarbazole-9-ethyl-carbonylhydazine (BCEC) is described and the spectrum properties of reagent itself is also studied. At the same time, the applications of the reagent for the analysis of real sample by HPLC are evaluated.2.1 The separation and determination of aldehydes using BCEC as labeling reagent by HPLC with fluorescence detection are evaluated. The derivatization conditions in conjunction with the gradiet elution are optimized. At the same time,the detection limit and linearity for derivatized aldehydes are obtained under the optimal conditions. The established method for the determination of aldehydes from alcoholic beverage is investigated with satisfactory results.2.2 The separation and determination of saccharides using BCEC as labeling reagent by HPLC with fluorescence detection are studied. The derivatization conditions in conjunction with the gradiet elution are optimized. At the same time, the detection limit and linearity for derivatized saccharides are obtained under the optimal conditions.2.3 The separation and determination of steroids using BCEC as labeling reagent by HPLC with fluorescence detection are evaluated.The derivatization conditions in conjunction with the gradiet elution are optimized. At the same time, the detection limit and linearity for derivatized steroids are obtained under the optimal conditions. The established method for the determination of steroids from rat feces is evaluated with satisfactory results.Chapter three: The synthesis of 2-[2-(toluene-4-sulfonate) ethyl]-pyrazolo[3,4-b]-7-methylquinoline are described. The applications of 2-[2-( toluene-4-sulfonate) ethyl]pyrazolo[3,4-b]-7-methylquinoline for the analysis of free fatty acids with HPLC are studied. The derivatization conditions in conjunction with the gradiet elution are optimized. At the same time, the detection limit and linearity for derivatized free fatty acids are obtained under the optimal conditions. The established method for the determination of free fatty acids from extracted bryophyte samples is evaluated with satisfactory results.更多还原