Fluorescent Probe Molecules Preparation And High Performance Liquid Chromatography

High performance liquid chromatography(HPLC) due to its separation characteristic has been used widely as an effective approach in many fields, such as chemistry , biology, toxicology, clinical sciences, environmental science and so on. It is also one of the fastest developing and the most widely used methods in analytical chemistry. However, not all compounds are accessible to be analyzed by HPLC directly due to the limitation of the compounds properties and the low content of corresponding components. Therefore, improving the sensitivity of detection is essential. As for the limitation of improving the detection sensitivity of instrument, developing novel fluorescent probe is a fruitful approach.Therefor, we synthesized three novel fluorescence reagents by different chemistry modification and investigated the spectrum properties in this thesis. By means of chemical derivatization, they were used to analyze carboxylic and amino target compounds by HPLC with satisfactory results. The thesis consists of five chapters.Chapter one: First, the fluorescence theory and chemistry derivatization were introduced systematically. Second, the development and the basic knowledge of HPLC were briefly introduced. Then, the applications of fluorescence reagents to carboxyl and amino compounds by HPLC were summarized.Chapter two: The synthesis techniques of three fluorescence reagents: 1,2-Benzo-3,4- Dihydrocarbazole-9-Ethyl-p-Toluenesulfonate(BDETS), l,2-Benzo-3,4-Dihydrocarbazole-9-Ethyl-Benzenesulfonate(BDEBS) and l,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate(BCEOC) are described, and followed by their structure token and spectrum properties.Chapter three: The applications of l,2-Benzo-3,4-Dihydrocarbazole-9 -Ethyl-p- Toluenesulfonate(BDETS)in the analysis with HPLC-MS were studied. This chapter consists of the analyses of fatty acids and bile acids. And by analyzing real samples, these methods were tested showing the feasibility.