Preparation Of A Novel 1,8-naphthalene Diimide Fluorescent Derivatization Reagent And Its Analytical Application In High Performance Liquid Chromatograph

High-performance liquid chromatography(HPLC)is widely used in pharmaceutical research,food safety,environmental testing,chemical,and life science research fields because of its fast and rapid analysis,high separation efficiency,and stable performance with developed detectors such as UV and fluorescence detectors.HPLC with fluorescence detection(HPLC-FLD)exhibited highly sensitive and low detection limits.Most compounds show neither natural UV absorption nor fluorescence.so it is very difficult to detect them,therefore,chemical derivatization is necessary to increase detection sensitivity and improve selectivity by pre-column or post-column HPLC derivatization methods.The chemical derivatization methods have great expansion and application in HPLC analysis by introducing luminescent groups into the analytes,the derivatization can make the derivatives have better UV or fluorescence properties for the detection of target compounds by HPLC.So far,many fluorescent derivatization reagents have been developed,and the popular fluorescent probes are mainly carbazole,acridine,coumarin,fluorescein,and rhodamine,while only a small part of fluorescent probes are developed based on 1,8-naphthalenediimide parent ring.In this work,three new fluorescent derivatization reagents based on 1,8-naphthalenediimide parent ring are designed,synthesized and characterized.Based on the properties of the tested target compounds,the optimized derivatization conditions are investigated for three new fluorescent reagents.The completed chromatographic separation,method validation and actual sample determination in combination with high-performance liquid chromatography with fluorescence detection are valuated.The main contents of this thesis consist of the following five chapters.Chapter one: This chapter summarizes three main parts: the research progress of high-performance liquid chromatography;the research progress of chemical derivatization techniques,and the research progress of naphthalimide fluorescencebased derivatization reagents.High-performance liquid chromatography with fluorescence detection exhibits high sensitivity and low detection limit.The rapid development of chemical derivatization technology has led to the further development and application of novel 1,8-naphthalenediamide-based fluorescence reagents.Chapter two: Three pre-column fluorescent derivatization reagents including4-[2-(benzoisoquinoline-1,3-dione)-ethyl]benzenesulfonyl chloride(BIEBS-Cl),4-((2-methyl-1,3-dioxo-2,3-dihydro-1H-benzo[de]isoquinoline-6-yl)oxy))-benzenesulfonyl chloride(MBIOBS-Cl),and 4-((2-methyl-1,3-dioxo-2,3-dihydro-1Hbenzo[de]isoquinoline-6-yl)amino)-ethyl-4-methyl benzenesulfonate(MNDMB)were prepared based on the 1,8-naphthalenediimide parent with a modification at the 4-and N-positions,and structurally characterized using IR,NMR,and MS.The spectral properties of BIEBS-Cl and MBIOBS-Cl were also investigated and compared.Chapter three: A precolumn derivatization method for the accurate determination of 11 fatty amines using BIEBS-Cl as the derivatization reagent by high-performance liquid chromatography with fluorescence detection was established.The method was successfully applied to the determination of 11 fatty amines in environmental water samples with satisfactory results.The method is of great significance for the evaluation of fatty amines in water samples.Chapter four: The precolumn derivatization methods for the accurate determination of four estrogens,six biogenic amines,and five phenols,respectively,using MBIOBS-Cl as the fluorescence derivatization reagent by high-performance liquid chromatography-fluorescence detection were developed.The corresponding methods were optimized for derivatization conditions,chromatographic conditions,and methodology verification,respectively,and applied to the determination of relevant practical samples with satisfactory results.Chapter five: The precolumn derivatization methods for the accurate determination of 15 saturated fatty acids,7 unsaturated fatty acids,and 4 aromatic acids,respectively,using MNDMB as the fluorescence derivatization reagent by highperformance liquid chromatography-fluorescence detection were established.The corresponding methods were optimized including derivatization conditions and chromatographic conditions.The established methods were successfully applied to the determination of relevant practical samples,and the experimental results showed the accuracy and sensitivity of the established method can meet the requirements for the determination of fatty acids in foods and benzoic acid in pharmaceuticals,which is of great significance for the quality evaluation of foods and pharmaceuticals.