Establishment Of Fingerprint Of Tianmeng Oral Liquid And Efficacy-related Components Study

Tianmeng oral liquid is composed of 17 kinds of traditional Chinese medicines and has the effects of nourishing qi and kidney,strengthening the spleen and stomach,nourishing the heart and tranquilizing mind.It is used to treat dizziness,tinnitus,visual impairment,hearing loss,insomnia,forgetfulness,loss of appetite,weak waist and knees,shortness of breath,and sequelae of stroke.It also has certain effects on cerebral dysfunction,coronary artery disease,cerebrovascular embolism,and hair loss.The current quality standard of Tianmeng oral liquid only stipulates one content measurement index of icariin,which is difficult to reflect fully the quality of Tianmeng oral liquid,so this paper used a variety of methods to conduct a comprehensive quality evaluation of Tianmeng oral liquid.Use single-column and column switching technology to establish HPLC fingerprint of Tianmeng oral liquid combined with multiple component quantitative analysis to evaluate comprehensively the quality of Tianmeng oral liquid.Use high-resolution liquid chromatography-mass spectrometry to study the chemical basis of Tianmeng oral liquid.Determine the components related to antioxidant activity in Tianmeng oral liquid through fingerprint-effect relationship research.Explore mechanism of action of Tianmeng oral liquid through network pharmacology.Through the research mentioned above,it provides a valuable reference for the enhancement and improvement of the quality standard of Tianmeng oral liquid.The researches of this paper mainly include the following parts:1.Establishment of HPLC fingerprint of Tianmeng oral liquidFirstly,the HPLC fingerprint of Tianmeng oral liquid based on a single C18 column was established and 12 batches of samples were determined.The 16 common peaks were matched by using ’Similarity Evaluation System for Chromatographic Fingerprint of TCM’.The similarity analysis and cluster analysis can accurately distinguish between expired and unexpired samples,realizing the quality evaluation of Tianmeng oral liquid.For traditional Chinese medicine preparations with complex components,the separation capability of a single column is limited,and it is difficult to achieve the desired separation effect.The column switching technology that connected C18 and C8 via six-way valve was used to establish the HPLC fingerprint of Tianmeng oral liquid.12 batches of samples were measured and 25 common peaks were matched.Compared with the fingerprint established by a single column,the separation effect was better and the peak capacity was higher.Similarity analysis and cluster analysis can be used to distinguish between expired and unexpired samples,providing a more effective method for the quality evaluation of traditional Chinese medicine preparations.2.Quantitative analysis of multi-components in Tianmeng oral liquidDeterminate syringin,chlorogenic acid,calycosin 7-O-glucoside,hesperidin and icariin in 25 batches of Tianmeng oral liquid using chromatographic method of fingerprint.Ferulic acid,formononetin,baohuoside I,vitexin-2’-O-rhamnoside,acteoside,isofraxidin,strychnine and brucine were determined by HPLC-MS method.The content of most of the components differed greatly between expired and unexpired samples,indicating that the quality of expired and unexpired samples was different.Combined with principal component analysis,the contents of components,such as baohuoside I,acteoside,and syringin,in the expired and unexpired samples were significantly different,which could be used as references of quality control indicators for Tianmeng oral liquid.3.Qualitative analysis of Tianmeng oral liquidUltra-high performance liquid chromatography-mass spectrometry was used to study the chemical composition of Tianmeng oral liquid.According to mass spectrometry data and literature information,96 components were identified,including 46 flavonoids,19 organic acids,8 triterpenoids,8 glycosides,4 amino acids,3 alkaloids and 8 other compounds,preliminarily clarifing the chemical basis of Tianmeng oral liquid.4.Correlation analysis of HPLC fingerprint and antioxidant activity of Tianmeng oral liquidThe antioxidant activity of 25 batches of Tianmeng oral liquid was measured by 1,1-diphenyl-2-picrylhydrazyl radical scavenging assays and Ferric reducing ability of plasma method.HPLC fingerprints of 25 batches of samples were determined.The correlation between fingerprints and two antioxidant activities was analyzed by orthogonal signal correction-partial least squares regression method.The components related to antioxidant activity of Tianmeng oral liquid were determined as hesperidin,catechol,and woodorien.5.Explore the mechanism of action of Tianmeng oral liquid by network pharmacologyThe 96 compounds identified in Tianmeng oral liquid were considered to analysis,and the active ingredients were selected according to oral bioavailability≥30%and drug-like≥0.18.By searching the databases,the 18 active ingredients of Tianmeng oral liquid mainly acted on 267 targets,and the topology analysis of the component-target network was performed.PTGS2,HSP90AB1,PTGS1,NCOA2,and CALM were the main targets of Tianmeng oral liquid.The protein-protein interaction(PPI)network included 218 targets,and MAPK1,MAPK3,TP53,AKT1，and JUN were the main targets of the PPI network,speculating that Tianmeng oral liquid directly or indirectly interacted with these targets to play a therapeutic effect.The 49 key targets of the component-target network of Tianmeng oral liquid were enriched in 19 GO functions and 64 KEGG pathways.Preliminary verification of the main targets and pathways through the literature information provided directions for further research on the mechanism of action of Tianmeng oral liquid.