New Fluorescent Derivative Reagent In The Application Of The Reversed Phase High Performance Liquid Chromatography - Mass Spectrometry

In recent years, High performance liquid chromatography(HPLC) has become one of themost famous method of separating mixtures because of its rapid, high sensitivity, selectivity. Ithas been successfully applied to medicine, food, environment, chemical industry and otherfields. Due to high performance liquid chromatography lack of a general-purpose detector aswell as the gas chromatography with flame ionization detector, the versatility of its detector isnot satisfactory. So we can not use the high performance liquid chromatography to nanalyze thesample directly, when the content of the sample is low or limited by its nature. In recent years,Many chemists use chemical derivative method to attempt to increase the sensitivity andseparation efficiency of the high-performance liquid chromatography. Although there has beenmany fluorescent reagents used in this method, they have different shortcomings in application,such as low sensitivity and so on. Designing fluorescent reagents to meet the actual needs has avery important practical application. The purpose of this paper is to synthesize and expand thescope of application of highly sensitive fluorescence derivatization reagents. In this paper, thefluorescence spectral properties, optimal derivatization conditions, chromatographic separationconditions were investigated. Pre-column derivatization reversed-phase high-performanceliquid chromatographic methods were established and evaluated for the analysis of realisticsamples.Chapter one: An overview of the development and basic theory about high performanceliquid chromatography and high performance liquid chromatography coupled with massspectrometry technique, introduced the principle and development about fluorescentderivatization, cited widely used derivative reagents in recent years.Chapter two: Design and synthesis of one novel fluorescent lableling reagent:2-(9H-carbazol-9-yl)propan-2-yl carbonochloridate. Then this compound was characterized andresearched by its structure and spectroscopic properties.Chapter three: Pre-column derivatization methods for the sensitive determination oftwelve kinds of aliphatic amine standards with2-(9H-carbazol-9-yl)propan-2-yl asderivatization reagent followed by HPLC/MS analysis have been developed. The optimalderivatization and chromatographic separation conditionsusing2-(9H-carbazol-9-yl)propan-2-yl as labeling reagents were evaluated. All derivatives wereseparated with a good baseline resolution and identified by MS. The established method for thedetermination of amines from real samples was satisfactory.Chapter four: Pre-column derivatization methods for the sensitive determination of sixkinds of aromatic amines standards with2-(9H-carbazol-9-yl)propan-2-yl as derivatizationreagent followed by HPLC/MS analysis have been developed. The optimal derivatization and chromatographic separation conditions were evaluated. The established method for thedetermination of aromatic amines from real samples sewage was satisfactory.Chapter five: Pre-column derivatization methods for the sensitive determination of sixkinds of phenolic compounds standards with2-(9H-carbazol-9-yl)propan-2-yl as derivatizationreagent followed by HPLC/MS analysis have been developed. The optimal derivatization andchromatographic separation conditions were evaluated. The established method for thedetermination of phenolic compounds from real samples textile was satisfactory.