Quality Control Of Pulsatilla Chinensis Based On Solid-phase Extraction And Protective Mechanism Of Pulsatilla Chinensis Against Liver Injury

Objective:Firstly,the SPE method with SBA-15 as the adsorbent was established,and UHPLC-CAD technology was used to enrich and determine the triterpenoid saponins in Pulsatilla chinensis.And then the in vitro and in vivo metabolism of hederasaponin C was studied.Finally,the network pharmacology method comprehensively decomposes the hepatoprotective effect of Pulsatilla chinensis.Methods:At first,the structure and morphology of the synthesized SBA-15 were evaluated by infrared spectroscopy,thermogravimetric analysis and transmission electron microscopy.UHPLC-CAD technology was used to evaluate the adsorption performance of SBA-15 on triterpenoid saponin compounds.The SPE method with SBA-15 as the adsorbent was established and validation of the method was carried out.The method was applied to the enrichment,separation and determination of triterpenoid saponins in Pulsatilla chinensis.Then,UHPLC/Q-Orbitrap-MS technology and Compound Discoverer?2.1 software were used to detect and process the hederasaponin C metabolites in vivo and in vitro.The structure of metabolites in vivo and in vitro was inferred based on the biotransformation of drugs and the cleavage pathway of hederasaponin C.Finally,the network pharmacology platforms including TCMSP,TCMIP,Pub Chem,Swiss Target Prediction Database,STICH database,Uni Prot,TTD,Pharm Gk and other databases were used to screen out active compounds targets of Pulsatilla chinensis,liver injury diseases.Meanwhile,the network and anlysis were established was performed.Results:The results of infrared spectroscopy,thermogravimetric analysis and transmission electron microscopy were consistent with those reported in the literature,indicating that the mesoporous molecular sieve SBA-15 was successfully synthesized.The optimized experimental conditions in this experiment were determined as follows:25 mg SBA-15 as a solid phase adsorbent,the p H was set to 5 and the elution solvent was 70%ethanol.Under the optimized conditions,the satisfactory linearity(r~2>0.9940)was acquired and the limits of detection reached 0.461-0.976?g/m L for the target analytes.The recoveries were ranged from 92.1%to 96.7%.A total of 34 metabolites were detected and tentatively identified by analyzing comprehensive biological samples including 26 phase I and 8 phase II metabolites.Using the network pharmacology database,19 active ingredient targets of Pulsatilla and16 core targets were screened.GO analysis was used to screen for information with significant difference P<0.05,22 biological processes,4 cellular component,3 molecular functional information.These mainly related to ABC transport and bile secretion pathway.Conclusion:In this study,the SPE method with SBA-15 as the adsorbent was established,and the UHPLC-CAD technology was applied to the enrichment and determination of triterpenoid saponins in Pulsatilla chinensis.It provides technical support for the study of trace triterpenoid saponins compounds in the complex system of traditional Chinese medicine and provides a theoretical basis for the application of mesoporous materials in solid phase extraction technology.It also studies the metabolites of in vivo and in vitro biological samples after oral administration of hederacoside C,providing valuable information for its pharmacological research and clinical application.This study revealed the hepatoprotective mechanism of Pulsatilla chinensis,and provides theoretical support and scientific basis for further research on the pharmacological mechanism of Pulsatilla chinensis.