Study On Quality Attributes Of Shenjie Granules Based On Mass Transfer Law

Objective:Shenjie granule is from the Qing Dynasty compiled by Yang Lishan’s’Shanghan Wenyi Tiaobian’-Shenjiesan.In this study,the preparation process of Shenjie Powder was studied,the quality standard of Shenjie Granules was formulated,the quantity and quality transfer law of Shenjie Powder decoction pieces-extract powder-granules was further studied,and its mechanism of action on epidemic diseases was explored,so as to predict the efficacy-related substances of Shenjie Powder,and provide reference for further exploration of the pharmacodynamic material basis and development and utilization of Shenjiesan.Methods:（1）By reading the literature,the records,clinical application status and frontier progress of chemical constituents of Shenjiesan were sorted out;the extraction process of Shenjie Granules was studied by orthogonal test:the extraction rate,extract,chlorogenic acid and baicalin content of Shenjiesan were used as evaluation indexes to determine the amount of water added,decoction time and times.Taking the forming rate,angle of repose,bulk density and moisture absorption rate as evaluation indexes,the entropy weight method was used to assign weights,and the star point-response surface method was used to optimize the best forming process of Shenjie granules;（2）The quality attributes of Shenjie Granules were explored by thin layer identification and high performance liquid chromatography,and their quality standards were established;（3）The fingerprint of Shenjiesan extract powder and granules was established by high performance liquid chromatography,and the content determination method was established to determine the four alkaloids such as baicalin and epiberberine in Shenjiesan extract powder and granules.The transfer rate of four alkaloids such as baicalin and epiberberine in Shenjiesan decoction pieces-extract powder-granules was calculated to clarify the mass transfer law of Shenjiesan decoction pieces-extract powder-granules;（4）TCMSP was used to screen the chemical SP to screen the chemical components of Shenjiesan,using the gene library search function to obtain the information of disease-related targets,using the STRING database to construct the PPI network for the selected main targets,and Cytoscape software was used to create a network diagram of the association between the active components of Shenjie Dissolution and epidemic diseases,and DAVID database was used to enrich GO and KEGG pathway information;Auto Dock Vina software was used to dock the key compounds obtained from network pharmacology with the core target proteins,and Pymol was used to visualize the docking results to confirm the binding degree of the potential active ingredients and targets of the compound.LPS was used to induce RAW264.7 cells to construct an in vitro inflammatory model.Enzyme-linked immunosorbent assay（ELISA）was used to detect the changes of pro-inflammatory factor TNF-α（tumor necrosis factor）in cell supernatant.Combined with fingerprint,network pharmacology,molecular docking and other related substances to predict the efficacy of Shenjie Dissolution.Results:（1）The preparation process of Shenjie Granules is as follows:add water with a solid-liquid ratio of 1:10（soak for 1.5 hours for the first time,add 2 times the amount of water）,decoction for 3 times,1 hour each time.Filtration,combined filtrate,concentrated to the thick paste,vacuum drying to dry extract,crushing,according to the drug ratio of 1:1.5 plus soluble starch,with 80%ethanol as wetting agent,mixing,making particles;（2）Establish the quality standard of Shenjie Granules:The traits of Shenjie Granules are light brown to brown granules with bitter taste;the specific identification method of Radix Scutellariae,Flos Lonicerae,Rhizoma Coptidis and Radix Platycodonis in Shenjie Granules was established.The granule inspection items all met the requirements;baicalin(C₂₁H₁₈O₁₁)in Baicalin shall not be less than 101.0mg per bag;（3）Study on the mass transfer law of Shenjie Granules:The fingerprints of 10 batches of Shenjiesan extracts and granules were established and 20chromatographic peaks were calibrated.Twelve chemical components were identified by reference substances,including berberine hydrochloride,chlorogenic acid,baicalin,isochlorogenic acid C,caffeic acid,quercetin,etc.The common peaks of the fingerprints of extract powder and granules were basically the same,and the similarity and correlation were good.The transfer rates of baicalin,coptisine hydrochloride,palmatine,berberine hydrochloride decoction pieces-extract powder and extract powder-granules were relatively consistent,and the transfer was stable.The preparation process of Shenjie Granules and the source of medicinal materials had certain stability and consistency;（4）By constructing a traditional Chinese medicine-component-target network to find out the relevant active components in Shenjiesan,it was found that quercetin,kaempferol,luteolin,baicalein,wogonin and other components may be the main active components of Shenjiesan.ALB,IL6,CASP3,HSP90AA1 and other targets are the key targets for Shenjiesan to treat epidemic diseases.Shenjiesan has the effect of inhibiting the release of pro-inflammatory factors,and its mechanism may be related to the regulation of TLR signaling pathway.Based on fingerprint,mass transfer,network pharmacology and molecular docking studies,it was found that 12 components such as baicalin,epiberberine,coptisine hydrochloride,palmatine,and berberine hydrochloride were closely related to the functional attributes of Shenjie Dissolution,which could be predicted as potential efficacy-related substances affecting the quality attributes of Shenjie Dissolution.Conclusion:The quality standard of Shenjie Granules was established,and the mass transfer law of Shenjiesan-extract powder-granules was studied.It was found that the mechanism of Shenjiesan on epidemic diseases was related to the regulation of inflammatory factor expression and immune regulation.The related substances of Shenjiesan were predicted,which provided reference for the pharmacodynamic mechanism,quality control and further development and utilization of Shenjie Granules.