The New Fluorescent Reagent For Aldehydes And Acids Determination

For the rapid analysis, high separation efficiency and automatization, high performance liquid chromatographic method has been widely used in a lot of fields, such as pharmaceutical analysis, biochemical analysis and environmental analysis. It is enthusiastically and initiatively adopted in many scientific fields for ultra strong separation ability, and turned into one of the fastest developing and widely used research field. However, not all compounds are accessible to be analyzed by chromatography directly due to the limitation of the compounds'properties and the low content of detected components. The components can be changed into derivatives by fluorescence derivatization to improve the detection sensitivity and, consequently, detected by fluorescence detector. So the development and the application of new fluorscence labeling reagents are the active research areas for chromatography. In this paper, two novel derivatization reagents were designed, prepared and characterized. The spectral properties and optimal derivatization conditions were investigated. The corresponding high performance liquid chromatographic methods were established and applied to analyse the real samples. The research contents of the paper are as follows:In chapter one, the basic theory about high performance liquid chromatography and high performance liquid chromatography coupled with mass spectrometry was simply presented. The chemical derivatization and fluorescence derivatization were introduced systematically. The applications of fluorescence reagents were summarized.In chapter two, the design and synthesis of fluorescence reagents, 2-(2-(4-(dimethylamino)-phenyl)-1H-phenanthro[9,10-d]imidazol-1-yl)ethyl-4-benzenesulfonate (PPMB) and 2-(9-oxoacridin-10(9H)-yl)acetohydrazide (AACE-NHNH2) were described, and followed with their structural characterization and spectral properties.In chapter three, fifteen saturated and unsaturated fatty acids and twenty saturated fatty acids were determined using PPMB as pre-column labeling reagent. The optimization of derivatization and chromatographic separation conditions using PPMB as labeling reagent was evaluated. The established chromatographic separation method for the determination of free fatty acids from several medicinal herbs was summarized. The proposed method showed wide linear range, excellent reproducibility, and simple derivatization for the determination of fatty acids.In chapter four, nine bile acids were separated and determined using PPMB as pre-column labeling reagent. The optimization of derivatization and chromatographic separation conditions using PPMB as labeling reagent was evaluated. The established chromatographic separation method for the determination of bile acids from human serum was summarized, and the results suggested that the method has great feasibility.In chapter five, ten aliphatic aldehydes were determined using AACE-NHNH2 as pre-column labeling reagent. The optimization of derivatization and chromatographic separation conditions using AACE-NHNH2 as labeling reagent was evaluated. The established chromatographic separation method for the determination of aliphatic aldehydes from human serum was summarized, and the results suggested that the method has great feasibility.