苯并咪唑类分子探针在液相色谱—质谱中的应用

【作者】 朱绪珍；

【导师】 尤进茂；

【作者基本信息】 曲阜师范大学 ， 有机化学， 2011， 硕士

【摘要】 高效液相色谱(High Performance Liquid Chromatography)因其强大的分离分析能力而成为分析实验室常用的仪器,已被广泛运用到许多科学研究领域如化学,生物化学,毒物学,临床医学,工业、农业、商检和法检、神经生物学、食品卫生等。高效液相色谱法是目前为止已经掌握的对复杂化合物分离的最有效手段之一。由于样品中待测组分自身的结构性质及低含量的限制,并非所有的化合物都可以直接用于液相色谱分析。目前,荧光检测被认为是最灵敏的检测手段之一,利用荧光标记技术使本身不发荧光的化合物(如脂肪胺、芳香胺、酚、脂肪醛等)转化成发射荧光的衍生物,从而实现高灵敏的荧光检测。本文重点研究三种新型荧光衍生试剂的合成和结构表征,并且考察标记试剂的光谱性质。通过对衍生、色谱分离条件的优化,建立了高效、灵敏的高效液相色谱-质谱分析方法,并将所建立方法用于实际样品的测定中。具体工作内容如下：第一章：概述色谱法的创立和发展历史以及高效液相色谱的基础知识及其发展过程；介绍荧光原理和化学衍生化法；总结目前广泛使用的荧光标记试剂,综述标记胺类、酚类、脂肪醛的荧光试剂。第二章：三种新型荧光试剂：1-(1-氢-咪唑)-2-(2-苯基-1-氢-菲[9,10-d]咪唑-乙酮(IPPIE)、2-(2-苯基-1-氢-菲[9,10-d]咪唑-乙酰肼(PPID)、2-(2-(10-蒽基)-苯并咪唑)-乙酰肼(ABID)的设计合成、结构表征、光谱性质的研究。第三章：新型荧光试剂IPPIE用于脂肪胺高效液相色谱-质谱的分析,通过对衍生化条件以及色谱分离条件的优化,建立了脂肪胺的柱前衍生化高效液相色谱荧光检测法,并把该方法运用到土壤样品的测定中,实现样品中脂肪胺的高灵敏检测。结果表明IPPIE在无催化剂条件下实现对脂肪胺的高选择性衍生,衍生反应条件温和,衍生效率高,线性范围宽,重现性好,操作简单,灵敏度高。第四章：荧光试剂IPPIE用于芳香胺和酚类化合物的高效液相色谱-质谱的分析,通过对衍生化条件以及色谱分离条件的优化,建立了芳香胺和酚类化合物的柱前衍生化高效液相色谱荧光检测法,并将建立的两种方法分别用于布料中芳香胺的检测和污水中酚类化合物的检测,实验结果表明所建立的方法具有很大可行性。第五章：新型荧光试剂ABID用于短链脂肪醛的高效液相色谱-质谱分析。通过对衍生条件和色谱条件的优化,建立了脂肪醛的柱前衍生高效液相色谱荧光检测法,实验结果表明,该方法衍生反应条件温和,衍生产率高,重现性好,实验操作简单,灵敏度高。并将其运用到白酒样品中脂肪醛的测定,取得满意的结果。更多还原

【Abstract】 High Performance Liquid Chromatographic (HPLC) method has been widely used in many fields of scientific research, such as chemistry, biochemistry, toxicology, clinical medicine, industry, agriculture, commodity and law inspection, neurobiology and food sanitation. So far, HPLC is one of the most capable methods for separating complex mixture usually used by human, however, not all of compounds are accessible to be directly analyzed by HPLC due to the limitation of the compound properties and the low contents of corresponding compounds. Fluorescence detection is considered to be one of the most sensitive detection methods. Now the methods for derivatization of aliphatic amines, aliphatic aldehydes, phenol with fluorescence labeling reagents have been widely adopted, and it can alter the physical and chemical properties of target compounds and make them easier to be analyzed. In this paper, we designed and synthesizd three new fluorescence reagents and evaluated the UV-VIS spectra of them. The derivatization, chromatographic resolution and mass spectrum condition were investigated and established highly effective and sensitive HPLC methods which were used for the analysis of real samples. Specific works are as follows:Chapter one:Summarize the found and AMD of chromatography, the Grundlagen and developing process of HPLC; Summarize the fluorescent reagent widely used now, and overview on the fluorescent reagents for determination of amines, phenols,aliphatic aldehydes.Chapter two:The synthesis of three fluorescence reagents:1-(1H-imidazol-1-yl)-2 (2-phenyl-1H-phenanthro) [9,10-d]imidazol-1-yl)ethanone (IPPIE) 2-2(phenyl-1H-phenanthro[9,10-d]imidazol-1-yl)acetohydrazide (PPID) 2-2-(anthracen-9-yl)-1H-benzo[d]imidazol-1-yl)acetohydrazide (ABID) were described followed with their structure representation and the study of spectral quality.Chapter three:A pre-column derivatization method for the sensitive determination of aliphatic amines using 1-(1H-imidazol-1-yl)-2(2-phenyl-1H-phenanthro)[9,10-d]imidazol-1-yl) ethanone (IPPIE) as labeling reagent followed by HPLC-MS analysis has been developed. The method was employed to the determination of aliphatic amines in soil sample. The derivatization and chromatographic condition using IPPIE as labeling reagent were investigated. The result exhibited IPPIE showed high selectivity for determination of the aliphatic amines without catalyst DMAP. The method showed wide linear range, excellent reproductcibility, simple derivatization procedure and high sensitivity.Chapter four:The pre-column derivatization methods for the sensitive determination of phenols and aromatic amines using 1-(1H-imidazollyl)2(2-phenyl-1H-phenanthro)[9,10-d]imidazol-lyl)ethanone (IPPIE) as labeling reagent have been developed. The derivatization and chromatographic condition were investigated. The methods were applied to the determination of the aromatic amines in fabric and phenols in sewage. The result showed that the established methods had great feasibility.Chapter five:The pre-column derivatization method for the sensitive determination of short-chain aliphatic aldehydes with 2-2 (anthracen-9yl) 1H-benzo[d]imidazol-lyl)acetohydrazide (ABID) as labeling reagent has been developed. The result showed the method had mild derivatization situation, high derivatization efficiency, the simple SPI and good reproducibility. The method was used in the determination of aliphatic aldehydes in white spirits sample and achieved a good result.更多还原