Preparation Of Novel Ester/Amide Stationary Phase For HPLC And Their Analytical Applications

High performance liquid chromatography is an important branch of analytical science,which is a technology used primarily for multiple components separation and detection.Chromatographic stationary phase is the primary factor that determines the separation properties of HPLC.Since the establishment of modern high performance liquid chromatograph,the development of novel stationary phase has been the focus of chromatographic researchers.Due to the increasing complexity of analytical objects in analytical work,chromatographic analysis is facing new challenges.Therefore,the development of novel stationary phase is of important practical significance.In this work,surface bonded flavone stationary phase,surface bonded C₁₈ modified flavone stationary phase,surface bonded Phenylglycine zwitterionic stationary phase and carboxylic acid type solid phase extraction material were prepared through thiol initiated surface polymerization.Poly?glycidylmethacrylate-divinylbenzene??poly?GMA-DVB??microspheres were prepared through seeded swelling polymerization method and it was used as substrate for the preparation of carboxylic acid type stationary phase for high performance liquid chromatograph.In chapter one,the development of high performance liquid chromatograph technology,the support materials and functional ligands of high performance liquid chromatograph were reviewed,which mainly included the preparation methods of silica matrix stationary phase.Furthermore,the development of thiol initiated surface polymerization method and its application in the field of preparation of stationary phase for HPLC were also reviewed.In chapter two,the core structure of flavones was synthesized and it was modified with methacryloyl chloride,the double bond modified flavone was immobilized onto the surface of MPS modified silica,the resulting silica material was applied as stationary phase for HPLC.The stationary phase was characterized by means of scanning electronic microscope,FT-IR,elemental analysis and etc.The results demonstrated that the functional ligands was immobilized onto the surface of MPS modified silica.Alkyl benzenes and poly aromatic hydrocarbons were used as hydrophobic solutes to evaluate the chromatographic performance of the prepared stationary phase,the results shown that the prepared stationary phase possess retention mechanism of hydrophobic and?-?interaction.It was also applied to separate flavonols and terphenyls,the results were satisfying.In chapter three,C₁₈ modified core structure of flavone was synthesized and that was double bond modified with methacrylic anhydride,then that was immobilized onto the surface of MPS modified silica.FT-IR,elemental analysis and other means were used to certify the bonding density and other information of the prepared stationary phase.The chromatographic performance of the prepared stationary phase was evaluated with alkyl benzenes and poly aromatic hydrocarbons as probe,the results demonstrated that the resulting stationary phase possess retention mechanism of hydrophobic and?-?interaction.At last,the packed column was used to separate terphenyls and naphthols and the results were satisfying.In chapter four,methacrylic anhydride modified Phenylglycine was synthesized and immobilized onto the surface of MPS modified silica through thiol initiated surface polymerization method.The resulting surface bonded Phenylglycine silica material was applied as zwitterionic stationary phase for HPLC.FT-IR and other means was used to certify the functional ligand was successfully immobilized onto the MPS modified silica.Alkyl benzenes,poly aromatic hydrocarbons,nucleosides and bases were used as test solutes for chromatographic performance evaluation of the prepared stationary phase under reversed mode and hydrophilic interaction mode,respectively.The zeta potential of the stationary phase at different pH and salt concentration was measured and the retention behaviors of the probe under different conditions were discussed.The packed column was also used to separate organic acids and sulfanilamides and that was compared with commercial C₁₈ and HILIC-Amide columns,the results shown that the prepared stationary phase possess better selectivity toward organic acids and sulfanilamides.In chapter five,Poly?glycidylmethacrylate-divinylbenzene?(poly?GMA-DVB?microspheres were prepared through seeded swelling polymerization method and it was used as substrate for the preparation of carboxylic acid type stationary phase for high performance liquid chromatograph.FT-IR,elemental analysis and other means were used to characterize relevant information of the resulting stationary phase.Alkyl benzenes and poly aromatic hydrocarbons were used as probe for chromatographic performance evaluation of the synthesized stationary phase under hydrophobic interaction mode.It was also applied to separate isomers of nitroanilines,cresols and catechols.In hapter six,4-methacrylamidobenzoic acid was synthesized and it was used as functional ligand of solid phase extraction material.The functional ligand was immobilized onto MPS modified silica through thiol initiated surface polymerization.The resulting silica material was characterized by FT-IR,elemental analysis and etc.A detection method of two kinds of tetracyclines in milk based on the packed solid phase extraction column was established.The washing solvents and elution solvents in solid phase extraction was optimized.The detection limits of the established method for oxytetracycline and doxycycline were 2 ng/mL and 10ng/mL,respectively.