The New Fluorescence Derivatization Reagent For Phenols And Acids Determination

For the powerful separation and analytical capacities, high performance liquid chromatographic method has been widely used in many fields of scientific research and application, such as analytical chemistry, clinical medicine, pharmaceutical chemistry, biochemistry, food hygiene and environmental monitoring. Because of lacking highly sensitive all-purpose detector in reversed-phase high-performance liquid chromatography, chemical derivatization was adopted by researchers to improve detection sensitivity and achieve baseline separation. Therefore, developing highly sensitive fluorescence derivative reagent to meet the actual needs of HPLC is of great significance. The dominant ideology of this paper is to design a new fluorescence derivatization reagent in order to enhance detection sensitivity and improve the shortcoming of existing reagents in actual analysis process. From some ways we carry out research work such as designation and preparation of the new tag reagents, identification of spectral properties, development of new derivatization reaction, optimization of the chromatographic separation conditions, investigating mass spectrometry quantitative and qualitative, analyzing actual sample and so on. Specific work is as follows:Chapter One:The developing progress about high performance liquid chromatography and high performance liquid chromatography coupled with mass spectrometry were simply presented. The principles of fluorescence and chemical derivatization were introduced. The widely used fluorescence derivatization reagents were summarized.Chapter Two:2-amino-5-ethylbenzo[b]acridin-12(5H)-one and (2-(dimethylamino)-12-oxobenzo[b]acridin-5(12H)-yl)methyl 4-methylbenzenesulfonate were described followed with their structural characterization.Chapter Three:Pre-column derivatization method for the sensitive determination of fatty acid compounds using 5-(2-oxo-2-(piperazin-1-yl)ethyl)benzo[b]acridin-12(5H)-one as labeling reagent followed by HPLC-MS analysis has been developed. The optimal derivatization was evaluated. Then, this method was applied to the determination of actual sample and exhibited great feasibility.Chapter Four:Pre-column derivatization methods for the sensitive determination of aliphatic acid and bile acid compounds with 2-(12-oxobenzo[b]acridin-5(12H)-yl)ethyl 4-methylbenzenesulfonate(BAETS) as labeling reagent followed by HPLC-MS analysis have been developed. The optimization of derivatization and chromatographic separation conditions using BAETS as labeling reagent has been evaluated. And the feasibility of this method has been identified by analyzing real samples.Chapter Five:Pre-column derivatization method for the sensitive determination of phenolic compound with 2-[2-(7H-dibenzo [a,g]-carbazol-13-yl)] isopropyl chloroformate (DBCPC-Cl) as fluorescence derivative reagent followed by HPLC-MS analysis has been developed. The optimization of derivatization and chromatographic separation conditions using the derivative reagents as labeling reagent has been evaluated. The proposed method showed wide linear range, excellent reproducibility, and simple derivatization for the determination of aliphatic amines.