Stable Isotope Labelled Derivatization UHPLC-MS/MS Is Used For Metabolite And Drug Analysis

Ultra performance liquid chromatography-tandem mass spectrometry?UHPLC-MS/MS?technique was applied in various fields of analysis.The technique combined the good separation ability of liquid chromatography with the high specificity detection of tandem mass spectrometry,and it was suitable for the analysis and detection of complex biological samples.Chemical derivatization can improve mass spectrometry ionization efficiency and detection sensitivity.However,in order to reduce the difference in mass spectrometry response from the complexity of the matrix and the variety of analytes,the isotope internal standard was added before mass spectrometry analysis.In general,most isotopic internal standards were difficult to obtain and synthesize,so stable isotope labeling derivatization becomes an alternative method,which improves the sensitivity and accuracy of the analysis.Complex matrices in biological samples and excess derivatization reagents from the derivatization process often lead to severe matrix interference and instrumentation damage to UHPLC-MS/MS detection.Therefore,it is necessary to perform effective sample preparation before instrument analysis.Magnetic dispersion solid phase extraction?MDSPE?based on graphene oxide and molecularly imprinted polymers as adsorbent utilizes an external magnetic field to separate the analyte from the matrix,and obtains good adsorption efficiency and fast separation ability.This paper introduced three new combined technologies and UHPLC-MS/MS was used to detect hydroxyl-containing cholesterol and its metabolites,triptolide,gastrodin and its metabolite p-hydroxybenzyl alcohol in rat microdialysates.The specific work was as follows:Chapter One:A brief introduction to the research progress of ultra performance liquid chromatography-tandem mass spectrometry,stable isotope labeling derivatization,dispersive solid phase extraction,graphene oxide and moleculariy imprinted technique in sample preparation.ChapterTwo:Apairofstableisotope-labeledderivatizationreagents d₀-/d₃-3-N-methyl-2'-carboxyrhodamine 6G?d₀-/d₃-MCR6G?was designed and synthesized to label the analytes.Magnetic graphene oxide was used as the adsorbent of dispersion solid phase extraction in order to adsorb and extract.And above technique were combined with ultra performance liquid chromatography-tandem mass spectrometry for the detection of hydroxyl-containing cholesterol and its metabolites in rat blood microdialyates.Chapter Three:The stable isotope-labeled derivatization reagent d₀-/d₃-MCR6G were used to label the analyte.The triptolide derivative was used as a template molecule to synthesize a magnetic molecularly imprinted polymer,which was used as an adsorbent to adsorb and extract the derivative.The pharmacokinetic study of triptolide in rat brain and blood microdialysates were determined by ultra performance liquid chromatography-tandem mass spectrometry.This method is sensitive and rapid,providing a new method of triptolide in drug action research.Chapter Four:A new magnetic molecularly imprinted material was synthesized using d₀-/d₃-3-N-methyl-2'-carbonyl chloride rhodamine 6G?d₀-/d₃-MCCR6G?and tyrosol derivatives as virtual template,while microwave-assisted stable isotope labeling derivatization-magnetic dispersionsolidphaseextractioncombinedwithultraperformanceliquid chromatography-tandem mass spectrometry were used to detect synchronously the pharmacokinetics of gastrodin and its metabolites p-hydroxybenzyl alcohol.