Analysis Of Chemical Constituents Differences Between Fritillaria Cirrhosa D. Don And Fritillaria Thunbergii And Its Metabolomics Based On UPLC-Q-TOF-MS/MS Technology

Fritillaria cirrhosa D.Don and Fritillaria thunbergii are typical traditional bulk medicinal materials,Fritillaria unibracteata Hsiao et KC Hsia is the main basal plant of C.The best varieties of the mother’s medicinal effects,they all have the effects of clearing heat and resolving phlegm,moistening the lungs and relieving cough,dispersing knots and reducing swelling,and have a long history of medicinal use.In clinical application,Fritillaria cirrhosa D.Don and Fritillaria thunbergii are used differently.Fritillaria cirrhosa D.Don is mainly used for clearing heat and moistening the lung,while Fritillaria thunbergii is mainly used for clearing heat and dispersing the lung.Modern pharmacology has also proved that the Fritillaria unibracteata Hsiao et KC Hsia in Fritillaria cirrhosa D.Don has better properties.The antitussive and expectorant effect is a treasure in the base source plant of Fritillaria.However,at present,the pharmacodynamic material basis of Fritillaria cirrhosa D.Don and Fritillaria thunbergii is not clear,and the pharmacodynamics of Fritillaria cirrhosa D.Don and Fritillaria thunbergii have the characteristics of"multi-component and multi-target".How to improve the pharmacodynamics of the two The basis and how to establish the quality control method of the two are also the biggest problems and the key points of the modernization development of Fritillaria cirrhosa D.Don and Fritillaria thunbergii,and the research on the chemical components of traditional Chinese medicine is a prerequisite for elucidating the pharmacodynamic substances,mechanism of action and clinical efficacy of traditional Chinese medicine.It is particularly important to find an efficient,sensitive,rapid and systematic analysis method.Therefore,in this study,UPLC-Q-TOF-MS/MS technology was used to rapidly analyze and identify the chemical constituents of Fritillaria cirrhosa D.Don,Fritillaria unibracteata Hsiao et K.C.Hsia,Fritillaria thunbergii and wild of Fritillaria cirrhosa D.Don in different regions of Qinghai.The method was used to analyze the difference in chemical composition of Fritillaria cirrhosa D.Don,Fritillaria unibracteata Hsiao et K.C.Hsia,and Fritillaria thunbergii,as well as the difference in chemical composition of wild of Fritillaria cirrhosa D.Don collected in Qinghai Guoluo Prefecture and Qinghai Yushu Prefecture in different regions,so as to enrich the The composition of the chemical components of Fritillaria cirrhosa D.Don,Fritillaria unibracteata Hsiao et K.C.Hsia,and Fritillaria thunbergii provides reliable supporting data for the study of the pharmacodynamic material basis of the three kinds of Fritillaria and the formulation of quality standards for the Fritillaria cirrhosa D.Don in different regions of Qinghai.In addition,this experiment is also based on the method of"traditional Chinese medicine plasma medicinal chemistry"to analyze and identify the prototype blood components and metabolites of the alcoholic extracts of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria thunbergii.Metabolic pathways of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria thunbergii to exert their medicinal effects,and the metabolic mechanism of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria thunbergii in vivo was discussed.The main results of this study are as follows:1.Difference analysis of chemical constituents of Fritillaria cirrhosa D.Don and Fritillaria unibracteata Hsiao et K.C.Hsia（ESI^-）For the first time,the ultra-high performance liquid chromatography-quadru pole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS）technique was used to comprehensively characterize the chemical components of Fritillari a cirrhosa D.Don and Fritillaria unibracteata Hsiao et K.C.Hsia in negative ion mode.Analysis,by comparing the cracking rules of chemical components,combined with accurate relative molecular weight,primary and secondary mas s spectrometry information,literature information and Pub Med,Pub Chem,Mass Bankd and other databases,a total of 45 compounds were identified,35 were derived from Fritillaria cirrhosa D.Don,36were derived from Fritillaria unibr acteata Hsiao et K.C.Hsia A total of 7 types of chemical constituents were id entified,including 15 alkaloids,13 fatty acids,8 phenylpropanoids（including 2lignans）,terpenoids There are 4 components,3 saponins,1 flavonoids,1 carb ohydrate;through the Principal Component Analysis（PCA）and Orthogonal Part ial Least Squares-Discriminant Analysis（OPLS-DA）.Four differential compound s,such as demissidine-3-o-β-D-glucopyeanosy（1→4）glucopyranosideand pinelliaic acid,provided reference for the quality control of Fritillaria cirrhosa D.Don and Fritillaria unibracteata Hsiao et K.C.Hsia and the research on the phar macodynamic materialbasis,and provided basic database construction data for l ater experiments.2.Difference analysis of chemical composition of Fritillaria cirrhosa D.Don in different regions of QinghaiData were collected in positive ion mode,and a total of 54 chemical constituents were identified,including 32 alkaloid constituents（9 steroidal alkaloids,23isosteroidal alkaloids）,8 fatty acid constituents,and nucleosides.5 compounds,1sugar compound,1 oxazolidine compound,1 amide compound,6 unidentified compounds,5 compounds were confirmed by comparison with reference substances,and 18 compounds were identified in this genus for the first time;Through PCA and OPLS-DA analysis,the differential compounds of 20 wild Fritillaria cirrhosa D.Don were collected in Yushu and Guoluo,Qinghai Province,including adenosine,delarine,demissidine-3-o-β-D-glucopyeanosy（1→4）glucopyranoside,Isoverticine.,cordiline,hupebenizine,ebeiedine,which were used for the quality control of Fritillaria cirrhosa D.Don,protection of wild resources,and the genuineness of Fritillaria cirrhosa D.Don in different regions of Qinghai.The research on the authenticity of Fritillaria cirrhosa D.Don in different regions of Qinghai provides a scientific basis.3.Difference analysis of chemical constituents of Fritillaria unibracteata Hsiao et K.C.Hsia,Fritillaria cirrhosa D.Don,and Fritillaria thunbergii（ESI⁺、ESI^-）Collecting data in positive and negative ion modes,a total of 72 chemical components were identified,including 44 alkaloids,12 fatty acids,4 nucleosides,4lignans,and 3 carbohydrates 1,1 oxazolidine,1 amide,1 amino acid,1 schiroidide,1other,9 unidentified compounds,32 compounds were found in this genus for the first time It was identified;through PCA and OPLS-DA analysis,a total of 28 differential compounds were obtained of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria cirrhosa D.Don,a total of 22 differential compounds were obtained of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria thunbergii,a total of 23differential compounds were obtained of Fritillaria thunbergii and Fritillaria cirrhosa D.Don,.The compound further improves the composition of the chemical components of the three kinds of Fritillaria,provides a reference for the study of the clinical application of the three kinds of Fritillaria,and is of great significance to the development and utilization of the three kinds of Fritillaria.4.Study on the metabolites of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria thunbergii in rat plasmaFor the first time,the UPLC-Q-TOF-MS/MS technology combined with UNIFI software and manual analysis was used to comprehensively characterize the prototype blood components and metabolites in the plasma of rats after gavage of the alcoholic extracts of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria thunbergii,a total of 5 prototype blood components of verticinone,perimisine,hupebenizine,sipeimine,Zhebeirine were obtained.A total of 33 metabolites were identified in the alcoholic extracts plasma group of Fritillaria unibracteata Hsiao et K.C.Hsia,and a total of 34 metabolites were identified in the alcoholic extracts plasma group of Fritillaria thunbergii.Among them,there were 6 unique metabolites in the alcoholic extract plasma group of Fritillaria unibracteata Hsiao et K.C.Hsia,consist of perimisine,Sipeimine-3-β-D-glucoside,Zhebeirine,Hupeheoside,hupebenizine,there were 7 unique metabolites in the alcoholic extract plasma group of Fritillaria thunbergii,consist of perimisine,Sipeimine-3-β-D-glucoside,Zhebeirine,Peimine,hupehemonoside.Pharmacokinetics study of Fritillaria unibracteata Hsiao et K.C.Hsia and Fritillaria thunbergii,and the elucidation of the mechanism of action has laid the foundation.