Development Of New Sample Pretreatment Technology And Its UHPLC-MS/MS And MALDI-TOF-MS Analysis Applications

Ultra-highperformanceliquidchromatographytandemmassspectrometry?UHPLC-MS/MS?has combined UHPLC super separation ability with MS/MS high sensitivity,accurate quantification ability.Due to the advantages of efficient separation ability,fast analysis and accuracy,low detection limit,it was widely used in biological sample metabolite analysis,environmental analysis,food safety,drug testing and other fields.Stable isotope label derivatization?SILD?can improve the separation of analytes and significantly improve accuracy and sensitivity with mass spectrometry detection.Compared with the traditional dispersed solid phase extraction methods,magnetic dispersion solid phase extraction?MDSPE?can achieved separate only by the external magnetic,without the complex operation such as filtration and centrifugation.Metal organic framework materials?MOFs?have the advantages of large specific surface area,adjustable pore size,and carrying functionalized groups,which have been highlighted in the field of biological sample pretreatment.Matrix assisted laser desorption/ionization time-of-flight mass spectrometry?MALDI-TOF-MS?has the characteristics of high sensitivity,low sample consumption,simple operation,high throughput,etc.,At the same time,it can be used to detect analytes in a wide mass range,providing a fast and efficient detection method for biomacromolecules?such as proteins and glycopeptides?.Functionalized MOFs material were used to selectively enrich the glycopeptides in biological samples for MALDI-TOF-MS detection,which has the advantages of fast,simple and high throughput.The specific work of this paper is as follows:Chapter One:Briefly describe the principles,characteristics and applications of UHPLC-MS/MS,MALDI-TOF-MS,chemical derivatization strategy,sample pretreatment strategy.Chapter Two:A sample pretreatment technique of SILD-MDSPE was designed,and the multi-reaction monitoring model of UHPLC-MS/MS was used to establish and verify the analysis method of various neurosteroids and their related metabolites in rat blood microdialysates.ChapterThree:Designedandsynthesizedd₀/d₃-6-N-methylrhodamine6G-N-hydroxysuccinimidyl formate as a stable isotope-labeled derivatization reagent,combined with MDSPE to detection neurotransmitters and their related metabolites in the rat brain microdialysates under the UHPLC-MS/MS multi-reaction monitoring mode.Chapter Four:A cysteine functionalized metal organic framework material?MIL-101?NH₂?@Au-Cys?was designed and synthesized for the enrichment and purification of vancomycin and norvancomycin in serum samples.The extract achieves high throughput detection of MALDI-TOF-MS.