Study On The Quality Of Herba Epimedii And Hawthorn Leaves By Using UHPLC-Q-TOF/MS

Objective:Quality and relatively quantitative methods were used to study the quality of Herba Epimedii and hawthorn leaves by ultra high-performance liquid chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry(UHPLC-PDA-Q-TOF/MS)Methods:Epimedium sagittatum(Sieb.et Zucc.)Maxim.,Epimedium myrianthum Stearn,and Epimedium pubescens Maxim,the main species of cultivation industry,were used as research objects in the first part of this paper.Firstly,the chemical components of different plant parts of Epimedium were analyzed and identified by UHPLC-Q-TOF/MS combined with UNIFI data analysis platform.Secondly,the different plant parts of three Epimedium species and cultivated,corresponding wild samples and other wild samples of three Epimedium species were analyzed by PCA and OPLS-DA.Finally,UHPLC-PDA(?=270nm)combined with heatmap visualization were used to determine the relative contents of chemical components in different plant parts of three Epimedium species and the relative contents of 12 flavonoids and total flavonoids in the cultivated,corresponding wild and other wild samples.In the second part of this paper,hawthorn leaves of different species(Crataegus pinnatifida Bge.and Crataegus pinnatifida Bge.Var major N.E.Br.)and different seasons(Crataegus pinnatifida Bge.Var major N.E.Br.)were studied as research objects.Firstly,UHPLC-Q-TOF/MS combined with UNIFI data analysis platform were used to analyze the chemical constituents in hawthorn leaves.Secondly,PCA and OPLS-DA were used to analyze the differences in hawthorn leaves of different species.Finally,the relative contents of five flavonoids and total flavonoids,three organic acids and total organic acids in hawthorn leaves of different seasons were determined by UHPLC-PDA(?=280nm),and the relative contents of six triterpenoid acids and total triterpenoid acids in hawthorn leavesof different seasons were determined by UHPLC-ELSD.Results:A total of 74 chemical components were identified or tentatively identified from different plant parts of Epimedium by UHPLC-Q-TOF/MS combined with UNIFI data analysis platform.The results of PCA and OPLS-DA showed that there were differences in chemical components of leaves,stems and rhizomes,and 21 components with VIP value greater than 7 were obtained and identified;there was little difference between the cultivated and its corresponding wild samples of three Epimedium species,which indicated that the chemical compositions of cultivated samples were consistent with the corresponding wild samples.The results of relative quantitative analysis showed that the relative contents of magnoline in stems and rhizomes was higher than that in leaves,most flavonoids with the parent skeleton of anhydroicaritin(type A)had a high content in the leaves,most flavonoids with the parent skeleton of demethylanhydroicaritin(type B)had a high content in the rhizomes,and the contents of most compounds in stems were low;the relative contents of twelve flavonoids and total flavonoids in cultivated and its corresponding wild samples were similar and the quality of cultivated Epimedium was more stable.A total of 58 compounds were identified or tentatively identified from hawthorn leaves by UHPLC-Q-TOF/MS combined with UNIFI data analysis platform according to the accurate molecular weight and fragment ion given by mass spectrometry.The results of PCA and OPLS-DA showed that there were differences in hawthorn leaves of different species,24 differential components with VIP value grater than 7 were obtained,and 20 of them were identified.In general,the compositions and contents of flavonoids in leaves of Crataegus pinnatifida Bge.were richer than that in leaves of Crataegus pinnatifida Bge.Var major N.E.Br.From April to November,the contents of five flavonoids in hawthorn leaves decreased first and then increased,which reached the highest level in November;among the six triterpenoid acids,the contents of 2 ?,3 ?,19 ?-trihydroxyursolic acid,3-O-trans-p-coumaroyltormentic acid and crataegolic acid increased gradually,which reached the highest in November.The content of corosolic acid increased firstly and then decreased,and reached the highest in October.The contents of ursolic acid and oleanolic acid decreased gradually,and the contents were highest in April;among the three organic acids,the contents of chlorogenic acid and piscidic acid decreased firstly and thenincreased,in which the content of chlorogenic acid was the highest in November and the content of piscidic acid was the highest in April.The content of para-hydroxybenzoic acid did not change much,and its content was highest in April.The relative contents of total flavonoids,total triterpenoid acids and total organic acids in hawthorn leaves of different seasons were the highest in November,which showed that the overall quality of hawthorn leaves was the best in November.Therefore,it is suggested to harvest hawthorn leaves in November.Conclusion:The qualitative and relative quantitative methods were established by UHPLC-PDA-Q-TOF/MS combined with UNIFI data analysis platform and PCA and OPLS-DA,which could be used to study the quality of Herba Epimedii.The results clarified the differences in the chemical composition of the leaves,stems and rhizomes of Epimedium,and proved the germplasm selection of the cultivated Epimedium was successful.The results provided a scientific basis for the cultivation,development and clinical application of Epimedium medicinal materials.The qualitative and relative quantitative methods were established by UHPLC-Q-TOF/MS combined with multivariate statistical analysis and UHPLC-PDA and UHPLC-ELSD to analyze the chemical composition of hawthorn leaves and comprehensively study the quality of different speices and different seasons of hawthorn leaves.The results not only clearly defined the differences of chemical composition in hawthorn leaves of different species but also accurately and intuitively reflected the change trend of relative contents of flavonoids,triterpenoid acids and organic acids in hawthorn leaves of different seasons,which provided a reference for the selection of source varieties and timely harvesting for medicinal hawthorn leaves.