Benzo Imidazole Molecular Probes In Liquid Chromatography - Mass Spectrometry

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.