The Synthesis Of N-containing Heterocyclic Fluorescent Labeling Reagent And Its Application In High Performance Liquid Chromatography - Mass Spectrometry

High performance liquid chromatography(HPLC),due to its powerful separation capacity, has been enthusiastically and widely adopted in many fields of scientific research and industry, such as biochemistry,toxicology,clinical sciences,chemical industry,clinical medicine, neurobiology,food hygiene,environmental science and quality control.It is also one of the fastest developing and the most widely used methods in analytical chemistry.However,not all compounds are suitable to be directly analyzed by HPLC because of the limitation of the compound properties and the very low contents of the corresponding compounds.So it is important to enhance the sensitivity of detection.Currently,fluorescence detection is still recognized to be one of the most sensitive detection technologies.Therefore,derivatization of analytes with fluorescence labeling reagents has been widely adopted.It can alter the chemical and physical properties of target compounds and make them easier to analyze.In this paper,we designed and synthesized two novel fluorescence labeling reagents.The optimal derivatization and chromatographic separation conditions were investigated.At the same time,fluorescence and UV-Vis spectra of labeling reagents and the suitability of the developed method for the analysis of real samples were evaluated.Chapter one:First of all,the HPLC theory and fluorescence labeling technology including pre-column derivatization and post-column derivation were introduced systematically.The history and basic theory about HPLC were also simply introduced.Secondly,some newly-emerged fluorescent labeling reagents and their application of the detection object were outlined,then,the applications of fluorescence labeling reagents for the labeling of carboxyl, organic amines by HPLC and their shortcomings in derivative process were summarized.In the end,the main research topics and their practical significance were clarified.Chapter two:The syntheses of two fluorescence labeling reagents 2-(2-(anthracen-10-yl) -1H-benzimidazol-1-yl) ethyl-p-toluenesulfonate(ABETS),2-[2-(7H-dibenzo[a,g]-carbazol-13 -yl)]isopropyl chloroformate(DBCPC-C1) were described,and followed by their structural characterization and spectral properties.Chapter three:Pre-column derivatization methods for the sensitive determination of long-chained and short-chained free fatty acids using 2-(2-(anthracen-10-yl)-1H-benzimidazol -1-yl) ethyl-p-toluenesulfonate(ABETS) as labeling reagents followed by HPLC-MS analysis have been developed.The optimal derivatization and chromatographic separation conditions using ABETS as labeling reagent were evaluated.And the feasibility of this method has been identified by analyzing the root of stellera chamaejasme.The results showed that this method had good reproducibility and higher detection sensitivity.Chapter four:Pre-column derivatization methods for the sensitive determination of aliphatic amines using 2-(11H-benzo[a]carbazol-11-yl) isopropyl chloroformate(DBCPC-C1) as labeling reagents followed by HPLC-MS analysis have been developed.The optimal derivatization condition using DBCPC-C1 as labeling reagent was evaluated.And the feasibility of this method has been identified by analyzing sewage sample.The results demonstrated this method with the advantage of simplicity,no need of phase transfer catalyst,injection without pre-treatment,and it had a higher detection sensitivity.