The Development Of New Fluorescent Reagents And High Performance Liquid Chromatography

High-performance liquid chromatography (HPLC), which has powerful capacity for the separation of complex samples, is widely used in the fields of scientific research and industry. However, the content of some components in the analyzed practical samples is too low to be detected, at the same time, some components are not suitable to be directly analyzed by HPLC due to their limitation of properties. So it is important to enhance detective sensitivity and to change the chemical and physical properties of target compounds to make them to be detected easily. Currently, fluorescence detection is still one of the most sensitive detection technologies. As most analytes cannot emit UV or fluorescence, it is necessary to derivatize analytes with labeling reagents. In this paper, we designed and synthesized three novel fluorescence reagents. Moreover the optimal derivatization conditions and separation conditions for the labeled target compounds by high-performance liquid chromatography with fluorescence detection were investigated.Chapter one: The development history and basic theory about HPLC are simply introduced. The fluorescence theory and chemistry derivatization are introduced systematically. The applications of fluorescence reagents are summarized.Chapter two: The syntheses of three fluorescence reagents: 1,2-benzo-3,4-dihydrocarbazole-9-ethanol, 1,2-benzo-3,4-dihydrocarbazole-9-acetic acid, 1,2-benzo-3,4-dihydrocarbazole-9- isopropyl chloroformate are described followed with their structural characterization.Chapter three: A highly sensitive detection method for fatty amines using 1,2-benzo-3,4-dihydrocarbazole-9- acetic acid as the derivatization reagent is developed.Chapter four: A highly sensitive detection method for fatty alcohols using 1,2-benzo-3,4-dihydrocarbazole-9-acetic acid as the derivatization reagent is developed.Chapter five: A highly sensitive detection method for fatty acids using l,2-benzo-3,4-dihydrocarbazole-9-ethanol as the derivatization reagent is developed.Chapter six: A highly sensitive detection method for fatty amines using l,2-benzo-3,4-dihydrocarbazole-9-isopropyl chloroformate as the derivatization reagent is developed.Chapter seven: A highly sensitive detection method for polyamines using l,2-benzo-3,4-dihydrocarbazole-9- isopropyl chloro formate as the derivatization reagent is developed.