Study On Quality Evaluation Method Of Inulae Flos And Hawthorn(Crataegus Cuneata) Fruits

Traditional Chinese medicine Inulae flos has a long history of application and high medicinal value.It has the efficacies of eliminating phlegm,relieving cough,relieving asthma and so on.Its medicinal site is dried flower heads.Inula japonica Thunb.of compositae is specified in the pharmacopoeia and extremely common,and it is distributed in most regions such as north China,northeast China,central China,and eastern China.The dried flower heads of Inula helianthus-aquatica C.Y.Wu ex Ling of compositae are also used as medicine in Yunnan,Sichuan and other regions,and it is common in some medicinal materials markets.Currently,the pharmacodynamic material basis of Inulae flos is not clear,and there is a lack of clear quality control indicators.The establishment of scientific and reasonable quality control criterion is the key to evaluate the quality of Inulae flos.Hawthorn(Crataegus cuneata)fruits is the dried and mature fruits of Crataegus cuneata Sieb.et Zucc.of rosaceae.It has the functions of promoting digestion,invigorating stomach,promoting blood circulation,removing blood stasis,lowering blood lipid and so on.It is widely used in improving cardiovascular diseases such as blood stasis and hyperlipidemia.At present,its pharmacodynamic substance basis is not clear,and its quality evaluation method are few.The discovery of function-related quality control criterion is the key to evaluate the quality of hawthorn(Crataegus cuneata)fruits.The research contents of this thesis are divided into four parts:1.The UHPLC-Q-TOF-MS/MS technology was adopted for identifying compounds in positive and negative ion modes based on the literature information and the reference substances.A total of 37 chemical components are identified from Inula japonica Thunb.,and33 chemical components are identified from Inula helianthus-aquatica C.Y.Wu ex Ling,most of which belong to organic acids,flavonoids and terpenes.This section preliminarily analyzed and studied the chemical substance bases of two kinds of Inulae flos,which laid the foundation for the subsequent research.2.The method of network pharmacology combined with molecular docking was adopted.The chemical composition-target-pathway network of Inula japonica Thunb.and Inula helianthus-aquatica C.Y.Wu ex Ling for the treatment of cough was constructed by using various biological related databases for exploring and predicting the pharmacodynamic mechanism of two kinds of Inulae flos in the treatment of cough.And the key targets were selected for molecular docking analysis.The results indicated that the components from Inula japonica Thunb.and Inula helianthus-aquatica C.Y.Wu ex Ling such as quercetin,luteolin,nepetin and so on may act on key targets such as AKT1,TNF,EGFR and so on to treat cough.3.The fingerprints of Inula japonica Thunb.and Inula helianthus-aquatica C.Y.Wu ex Ling was established by using 0.1% formic acid aqueous solution and acetonitrile as the mobile phase firstly.Six compounds including chlorogenic acid,isoquercitrin,1,5-dicaffeoylquinic acid,isorhamnetin-3-O-glucoside,quercetin,luteolin were screened out as discrimination markers by using OPLS-DA analysis method.The discrimination model of Inula japonica Thunb.and Inula helianthus-aquatica C.Y.Wu ex Ling was established by using discriminant analysis based on the contents of six markers,so as to distinguish the two types of samples from the perspective of chemical composition.It provides ideas and basis for improving the quality evaluation system of Inulae flos.4.A strategy of metabonomics combined with chemometrics was established to screen function-related quality markers of hawthorn(Crataegus cuneata)fruits.Multiple batches samples were analyzed by UHPLC-Q-TOF-MS/MS and tested for thrombin activity inhibition.Seven compounds including D-(-)-quinic acid,procyanidin C1,orientin,rutin,isoquercitrin,quercetin,(+)-balanophonin were screened as function-related quality markers by using a variety of technical means such as OPLS-DA,molecular docking,Ada Boost algorithm,KNN algorithm,LC-MS/MS,discriminant analysis and so on.Thereby,the activity of different batches of hawthorn(Crataegus cuneata)fruits could be evaluated,which furnish a scientific basis for establish a quality evaluation system of hawthorn(Crataegus cuneata)fruits.