Improve The Quality Standards Of Shujin Huoxue Capsule Based On Quality Markers

Shujin Huoxue Capsule is a Traditional Chinese medicine preparation,consisting of Flos Carthami,Cyperus rotundus,Cortex Periplocae,Viscum coloratum,Lycopi herba,Rhizoma Cibotii,Trachelospermum jasminoides,Common clubmoss herb Latin,Kadsurae Caulis and Pyritum.It can prevent and treat the pain of muscle and bone,promote blood circulation,dissipate blood stasis,and inhibit platelet aggregation.Shujin Huoxue Capsule was developed according to the changes in the dosage form of Shujin Huoxue Tablet,which aimed to reduce the dosage of preparations,improve bioavailability,increase the efficacy,and facilitate the clinical use of patients.In the current quality standards of Shujin Huoxue Capsule,thin-layer chromatography(TLC)methods was established to qualitatively identify of Cyperus rotundus,Flos Carthami and Cortex Periplocae,High Performance Liquid Chromatography(HPLC)method was applied to determine the content of4-methoxysalicylaldehyde.However,the content of 4-methoxysalicylaldehyde in the raw medicinal materials was > 0.46%,which exceeded the standard of “Chinese Pharmacopoeia” for the raw medicinal Cortex Periplocae(the content of4-methoxysalicylaldehyde in the raw medicinal was > 0.20%).As a result,the Cortex Periplocae medicinal on the market was difficult to meet the requirement of production.Therefore,considering these problems existing in the current standard of Shujin Huoxue Capsule,it was urgent to establish a scientific and comprehensive quality standard to improve product quality.The following studies were carried out in this project,including HPLC fingerprint chromatogram,network pharmacology,quality markers and quality standard.(1)Establishment of HPLC fingerprint combined with chemical pattern recognition.Firstly,the HPLC analytical method was optimized,and ten common peaks of the fingerprints was confirmed.Then,the methodology was investigated,consisting of repeatability,precision and stability.Seven common peaks in the fingerprint were identified using HPLC-QTOF-MS/MS technology.The similarity of23 batches of Shujin Huoxue products(Shujinhuoxue capsules,Shujinhuoxue tablets)from different factories was evaluated.and combined with chemical pattern recognition methods such as principal component analysis,cluster analysis and orthogonal partial least squares discriminant analysis to achieve the overall quality control of Shujin Huoxue products.Fingerprints combined with chemical pattern recognition methods could effectively improve the overall quality control of traditional Chinese medicine.It was more suitable for the quality evaluation of complex systems of traditional Chinese medicine.(2)Study on the mechanism of osteoporosis based on network pharmacology.collected active compounds of ten herbs in Shujin Huoxue capsule using TCMSP database,and obtained common targets of active ingredients and disease using Genecards database.Then,all obtained common targets were separately imported into the Uni Prot database for normalization protein name and Gene ID,using DAVID database to screen important pathways,and using cytoscape3.6.1 software to obtain the “medicine-compound-target-signal pathway” network relationship.As a result,sixteen active compounds related to osteoporosis were screened as candidate components of quality markers.(3)Selection of new index components based on the five principles of quality markers.according to the five requirements in the search for quality markers,measurability,compatibility environment,transmission and traceability,effectiveness and uniqueness were considered to made a Cobweb Pattern Map.The top 5 quality markers in the comprehensive ranking were selected as candidate index components,including eleutheroside B,tracheloside,hydroxysafflor yellow A,formononetin,4-methoxysalicylicaldehyde.Where 4-methoxysalicylicaldehyde was the original index component.The liquid phase conditions of the other 4 candidate index components were optimized,finally tracheloside was selected as a quality marker in the new quality standard.(4)Establishment of new standards for Shujin Huoxue Capsules based on quality markers.TLC was used to qualitatively identify of tracheloside.The methodology of TLC and the determination of HPLC content were investigated.Then,optimizing the content determination method of the original index component4-methoxysalicylaldehyde.The differences in the content of4-methoxysalicylaldehyde in Shujin Huoxue products from different factories were investigated to determine the content limits.Finally,a relatively complete quality standard for Shujin Huoxue capsule was established.