Qualitative And Quantitative Analysis Of Major Constituents In Roots Of Millettia Speciosa Champ.by Uplc-Qtof-Ms/Ms And Uplc-Ms/Ms

The root of Millettia speciosa Champ.?Leguminosae?is one of the well-known traditional Chinese medicines abundant in phenolic compounds and plays important roles in treatment of pain or numbness of the joints,chronic bronchitis and chronic hepatitis.In this thesis,an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry?UPLC-QTOF-MS/MS?based chemical profiling approach was established for rapid separation and characterization of multi-constituents in roots of Millettia speciosa Champ.Furthermore,8 constituents,namely gentistic acid,salicylic acid,ononin,liquiritigenin,calycosin,formononetin,isoliquiritigenin and maackiain were simultaneously quantified by the developed ultra-high performance liquid chromatography coupled with a triple quadrupole mass spectrometry?UPLC-MS/MS?method.Two main contents of research section were as follows:Firstly,a validated method based on UPLC-QTOF-MS/MS was established to separate and identify compounds in Millettia speciosa Champ.?Leguminosae?.A total of 48 components were simultaneously detected,of which 38 components,including 21 isoflavones,4 phenolic acids,4 isoflavanones,2 phenolic aldehydes,2 flavanones,1 aliphatic acid,1 chalcone,1 flavonol,1 isoflavane and 1 pterocarpan were unambiguously identified or tentatively assigned based on the retention time,UV spectra,characteristic molecular ions,MS/MS fragmentation data and reference standards.The major diagnostic fragment ions of components were observed and the corresponding fragmentation pathways were proposed.The results showed that the loss of a 44 Da?-CO₂?could be considered as the characteristic fragment behavior of phenolic acids;the characteristic fragment ions from RDA reactions in C-ring were occurred at isoflavonoids;flavanones and chalcones all tend to undergo RDA fragmentation to produce ^1,3A^-ion as the base peak in negative ion mode.This investigation provides important analysis data for further quality control,pharmacological and toxicological research on Millettia speciosa Champ.Secondly,a simple and specific UPLC-MS/MS method was developed for the analysis of 8 bioactive components?gentistic acid,salicylic acid,ononin,liquiritigenin,calycosin,formononetin,isoliquiritigenin and maackiain?in Millettia speciosa Champ.The quantitative method was established based on multiple reactions monitoring?MRM?mode in ESI positive and negative ion modes and eight effective components were baseline separated in 8 min.The method showed good linear correlation?R²?0.9909?over a wide range and a high sensitivity.The average recoveries of the eight marker components were in the range of 90.7%?123.1%.As a result,the analytical method possessing high throughput and sensitivity is suitable for the quality control of the Millettia speciosa Champ.