Sulfonyl Chloride Type Fluorescent Derivative Reagent Synthesis And Their Application In Hplc/ms

In the field of high performance liquid chromatography (HPLC), chemical derivatization isadopted by researchers to achieve baseline separation and improve detection sensitivity as aneffective method. With the high sensitivity of fluorescence detection, developing new fluorescentderivatization reagents and its application in liquid chromatography is one of the active researchareas for chromatography. However, not all compounds are accessible to be analyzed bychromatography due to the limitation of the compounds’ properties and the low content ofdetected components. These compounds which are derivatived with fluorescent derivatizationreagents can detect by the fluorescence detection. The method can meet routine and trace sampleanalysis and has been widely used in many fields of scientific research, such as analyticalchemistry, food hygiene environmental monitoring and biochemistry. The purpose of this paperis to synthesize and expand the scope of application of highly sensitive fluorescencederivatization reagents. The chromophore of new fluorescence derivatization reagents is acridoneto improve the sensitivity of fluorescence detection and the the active group is chloride to reactwith many compounds. The new fluorescence derivatization reagents can be used to detect andanalysis amines and hormonal compounds, such as aiphatic amines, biogenic amines, aromaticamines, amino acids,estrone, estradiol and esreiol. In this paper, the UV and fluorescencespectral properties, optimal derivatization conditions, chromatographic separation conditionswere investigated. Pre-column derivatization reversed-phase high-performance liquidchromatographic methods were established and evaluated for the analysis of realistic samples.Chapter one：The developing progress and basic theory about HPLC coupled with massspectrometry (MS) were simply presented. The principles and applications of fluorescencederivatization and fluorescence derivatization reagents were introduced. The widely usedfluorescence derivatization reagents were summarized.Chapter two：Three fluorescence derivatization reagents were synthesized and characterized.They were10-phenyl-acridone-2-sulfonyl chloride (PASC),10-methyl-acridone-2-sulfonylchloride (MASC) and10-ethyl-acridine-2-sulfonyl chloride (EASC). The synthetic routes andderivatization schemes of acridone derivatives were summarized and compared. Their relation ofstructures and the UV and fluorescence spectral properties were also studied.Chapter three：Pre-column derivatization methods for the sensitive determination of twelvekinds of aliphatic amine standards with PASC as derivatization reagent followed by HPLC/MSanalysis have been developed. The optimal derivatization and chromatographic separationconditions using PASC as labeling reagents were evaluated. All derivatives were separated witha good baseline resolution and identified by MS. The established method for the determination of amines from real samples was satisfactory. Such as aliphatic amines in environmental water andsoil samples were determined. Compared with EASC, PASC derivatives show higher sensitivityin UV and fluorescence intensity.Chapter four：Pre-column derivatization methods for the sensitive determination of eightkinds of biogenic amine standards with PASC as derivatization reagent followed by HPLC/MSanalysis have been developed. The optimal derivatization and chromatographic separationconditions using PASC as labeling reagents were evaluated. All derivatives were separated witha good baseline resolution and identified by MS. The established method for the determination ofamines from real samples was satisfactory. Such as biogenic amines in canned sardines,authentic sausage, fresh pork, dry casein and beer were determined.Chapter five：Pre-column derivatization methods for the sensitive determination of threekinds of estrogens with PASC as derivatization reagent followed by HPLC/MS analysis havebeen developed. The optimal derivatization and chromatographic separation conditions usingPASC as labeling reagents were evaluated. All derivatives were separated with a good baselineresolution and identified by MS. This method can be used to measure estrogens in real samples.Chapter six：10-methyl-acridone-2-sulfonyl chloride (MASC) and d3-10-methyl acridone(d3-MASC) as a pair of novel isotope-labeled derivatization reagents were synthesized andcharacterized. These reagents provided a relative quantification method of labeling estrogens byHPLC and atmospheric pressure chemical ionization mass spectrometry (APCI-MS).This worklaid the foundations for relative quantitative analysis of estrogen in actual samples.