Fluorescent Reagent Preparation And Its Application In The Application Of The Reversed Phase High Performance Liquid Chromatography - Mass Spectrometry

HPLC is an important analysis and detection methods, get extensive research andapplication development increasingly, has already deep into many industry and research anddevelopment areas. Such as, drug comprehensive development, the petroleum chemical industry,food safety, environmental regulation, medical research and development, biological chemicalpreparation, separation and preparation, quality testing. Combined with HPLC and chemicalderivative method, will convert stay test sample components into fluorescence emission groups,change the tested samples chemical properties, using fluorescence detection of high sensitivityanalysis chemistry derivatives. Thus, improve the performance liquid chromatography analysispracticality. Therefore, the development of high performance design synthetic fluorescent markerreagent also become a derivative of high performance liquid chromatography auxiliary methodeffective in achieving fast and accurate, high sensitivity, automatic detection in an effective way.Based on the synthesized derivative reagents, prepared three new fluorescent marker reagents,investigated its spectral properties, and by using composite fluorescence properties establishedits characteristics of RPHPLC. At the same time, optimize the derivative conditions，explorechromatographic separation conditions and mass spectrum identification, and applied to actualsample to detect, etc. Specific content as follows:Chapter One: Review the HPLC/MS method, introduced the chemical derivative methodand FD, lists the common fluorescent derivative mark reagents.Chapter Two: Three kinds of fluorescent marker reagent synthesis:2-(2-(anthracen-10-yl)-1H-benzo[d]imidazol-1-yl)-1-(1H-imidazol-1-yl)ethanone(ABIE),4-(1-4-(1-methyl-1H-phenanthro[9,10-d]imidazol-2-yl)benzene amine(MPIA),10-(2-(1H-imidazol-1-yl)-2-oxoethyl)acridin-9(10H)-one(IEA), create structure characterizationand the determination of optical properties.Chapter Three: Fluorescent marker reagent:2-(2-dibenzo carbazole ethyoxyl)-chloroethane formiate. By HPLC/MS analysis tabor goldester. Through the optimization derivative conditions and chromatographic separation conditions,establish the tabor gold ester of Pre-column derivatization high performance liquidchromatographic separation and analysis methods. And used in the practice of the test samples,the results prove the methods to set up the actual operation feasibility is higher, derivative highefficiency.Chapter Four: Fluorescent marker reagent:2-(2-(anthracen-10-yl)-1H-benzo[d]imidazol-1-yl)-1-(1H-imidazol-1-yl)ethanone(ABIE)and10-(2-(1H-imidazol-1-yl)-2-oxoethyl)acridin-9(10H)-one(IEA), used for fat amine high performance liquid chromatography-mass spectrometry analysis. Through optimizing thederivative conditions and chromatographic separation conditions, create a fat amine HPLCanalysis method. Used in the practice of the test sample fat amine, results show that the methodhas a lot of feasibility, the experiment of simple operation, high resolution.Chapter Five: Fluorescent marker reagent:4-(1-4-(1-methyl-1H-phenanthro[9,10-d]imidazol-2-yl)benzene amine(MPIA), used forfatty acids in the HPLC spectrometry analysis. Through optimizing the derivative conditions andchromatographic separation conditions, establish the fatty acids in the column former derivativesHPLC analysis method. The experiments prove the feasibility of the method.