摘要
高效液相色谱(HPLC)是现代分离测定的重要手段,应用于工业和研究领域的各个方面,极大地推动了化学学科的发展。然而,因为缺少通用的检测器及样品被测组分本身性质及其低含量的限制,并非所有化合物都可直接用于液相色谱分析。为此利用化学衍生反应改变被测化合物性质使之能够被灵敏、快速和自动化测定具有重要意义。本文重点研究了三种新型荧光标记试剂的合成和表征,优化了四种衍生试剂的衍生反应条件、色谱分离条件,并将建立的方法应用于不同样品的分析。具体工作如下：
第一章：首先介绍了高效液相色谱的原理及化学衍生法中的柱前衍生和柱后衍生。其次综述了近年来出现的一些商品荧光标记试剂和本实验室合成的荧光试剂。简述了液质联用接口的发展过程。最后阐明了本课题的主要研究内容及其实际意义。
第二章：设计并合成了三种荧光标记试剂：2-[2-(7H-二苯并[a,g]咔唑)]-异丙基-4-甲基苯磺酸酯、1-(9-蒽甲基)-3-甲基吡唑啉酮、2-[12,13-二氢-(7H-二苯并[a,g]咔唑)]-乙醇氯甲酸酯。并对其结构进行了表征。
第三章：将荧光标记试剂4-(1H-菲并[9,10-d]咪唑-2-)苯甲酸(PIBA)用于酚的高效液相色谱-质谱分析。通过对衍生化条件和色谱分离条件的优化,建立了酚的柱前衍生高效液相色谱荧光检测法。并将该方法用于废水样品中酚的测定,检验了所建立的分析方法的可行性。结果表明此方法具有良好的重现性和较高的检测灵敏度。
第四章：将荧光标记试剂4-(1-甲基-1H-菲并[9,10-d]咪唑-2-)苯甲酸(MPIBA)用于芳香胺的高效液相色谱-质谱分析。通过对衍生化条件和色谱分离条件的优化,建立了芳香胺的柱前衍生高效液相色谱荧光检测法。并将该方法用于污水中芳香胺的测定,检验了所建立的分析方法的可行性。研究结果表明,此方法简单易行,检测灵敏度高。
第五章：将荧光标记试剂2-[12,13-二氢-(7H-二苯并[a,g]咔唑)]-乙基氯甲酸酯用于脂肪胺的高效液相色谱分析。通过对衍生条件的优化,建立了脂肪胺的柱前衍生高效液相色谱的荧光检测法。
第六章：将衍生试剂BCETs用于脂肪酸的高效液相色谱-质谱分析。建立了脂肪酸的柱前衍生色谱荧光检测法,并将该法应用于微孔草籽油中脂肪酸的测定。结果表明此方法能够实现微孔草籽油中脂肪酸的分析测定。
High performance liquid chromatography (HPLC), as one of the most important modern analytical means of separation and determination, has been used in all aspects of industrial and research fields and greatly promoted the development of chemistry. However, not all compounds are suitable to be directly analyzed by HPLC simply due to lack of general-purpose detector, the limitation of the compounds'properties and the low contents of the detected component. So it is significant to develop testing methods with high sensitivity. Chemical derivatizations is one of the most effective procedures to change the properties of corresponding compounds. In this paper, we designed and synthesized three novel fluorescence labeling reagents. The optimal derivatization and chromatographic separation conditions of labeling reagents were investigated. At the same time, fluorescence and UV-Vis spectrum of labeling reagents and the suitability of the developed method for the analysis of real samples were evaluated.
Chapter one:The basic theory about high performance liquid chromatography and pre-/post-column derivatizations were simply presented. Subsequently, commercial fluorescent reagent and the as-synthesized reagents of our laboratory were summarized. The process of developing interface about HPLC-MS was also presented. Lastly, the main works of the research and their practical significance were clarified.
Chapter two:Designed and synthesized of three novel fluorescence labeling reagents: 2-[2-(7H-dibenzo [a,g]-carbazol)]-isopropyl-4-methylbenzenesulfonic acid; 1-(9-methyl anthracene)-3-methyl pyrazolone; 2-[12,13-dihydrogen-(7H-dibenzo[a,g]-carbazol)]-ethanolchloroformate. Then, these reagents were described and followed by their structural characterization.
Chapter there:Pre-column derivatization method for the sensitive determination of phenolic compounds using 4-(1H-phenanthro[9,10-d]imidazol-2-yl)benzoic acid (PIBA) as labeling reagents followed by HPLC-MS analysis have been developed. The optimal derivatization and chromatographic separation conditions using PIBA as labeling reagent were evaluated. And the feasibility of this method has been identified by analyzing the drain water. The results showed that this method had good reproducibility and higher detection sensitivity.
Chapter four:Pre-column derivatization method for the sensitive determination of aromatic amine using 4-(1-methyl-1H [9,10-d]imidazol-2-yl)benzoic acid (MPIBA) as labeling reagents followed by HPLC-MS analysis have been developed. The optimal derivatization and chromatographic separation conditions using MPIBA as labeling reagent were evaluated. And the feasibility of this method has been identified by analyzing the drain water. The results showed that this method had simple and high detection sensitivity.
Chapter five:Pre-column derivatization method for the sensitive determination of aliphatic amine using 2-[12,13-dihy4rogen-(7H-dibenzo[a,g]-carbazol)]-ethanol chloroformate as labeling reagents followed by HPLC analysis have been developed. The optimal derivatization and chromatographic separation conditions using 2-[12,13-dihydrogen-(7H-dibenzo[a,g]-carbaz ol)]-ethanol chloroformate as labeling reagent were evaluated.
Chapter six:Pre-column derivatization method for the sensitive determination of aromatic amine using BCETS as labeling reagents followed by HPLC-MS analysis have been developed. And the feasibility of this method has been identified by analyzing the Microula sikkimensis seed oil. The results showed that this method can be used to determination of fatty acids.
引文
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