Study On The Active Constituents Of Fupo Ganmao Granules By LC-MS And Evaluation Of Its Anti-inflammatory Activity

The constituents of traditional Chinese medicines(TCMs)are not usually clear due to the complicated components and low content.Fupo Ganmao(FPGM)Granules,which are composed of four herbal medicines,Hibisci Mutabilis Folium,Magnolia officinalis,Pericarpium Citri Reticultae,Arctii Frutus,is a regularly used traditional TCM for the treatment of common colds.However,the unclarity of chemical composition of this formulation restricted its basic research on effective substance.In order to identify the active components from FPGM,components was rapid identified by HPLC-IT-MSn based on the mass spectrometric fragmentation feature.Three kinds of solvent were involved in the extract procedure of crude herbs,and then series of components and few compounds were identified for the first time.In addition,anti-inflammatory activity of FPGM and main constituents have been investigated.The main contents and results were described as follows:1.The qualitative study of comprehensive constituents of FPGM.The HPLC-IT-MSn method was established for rapid isolation and identification of FPGM.As a result,a total of 117 compounds were detected from the FPGM,and 72 compounds were unambiguously identified or tentatively characterized by comparing the retention time,mass spectrometry fragmentation feature,including 25 lignans,23 flavonoids,15 phenolic acids,4 alkaloids and 5 other compounds.The component herb from which each compounds was derived was also analyzed.The chemical composition of FPGM was comprehensively and systematically analyzed,and the fragmentation of flavonoids,phenolic acids,lignans and alkaloids were studied and summarized.This part developed a comprehensive understanding of the multiple chemical constituents in FPGM and could offer chemical basis for the future study.2.Comparison of different extraction methods of single Chinese medicinal materials:water extraction,95%ethanol extraction and ethyl acetate extraction were used to extract four kinds of Chinese herb respectively,and then separated and identified by optimized HPLC-IT-MSn method.As a result,30 compounds from Hibisci Mutabilis Folium were identified,of which 8 were measured for the first time;38 compounds from Magnolia officinalis were identified,of which 7 were measured for the first time;75 compounds from Pericarpium Citri Reticultae were identified,including 11 compounds measured from the plant for the first time;55 compounds were identified from arctiin,and 9 of them were measuredfor the first time.3.Studies of in vitro anti-inflammatory effects:The in vitro anti-inflammatory effects of FPGM and its active constituents were investigated in LPS-stimulated RAW 264.7 cells.In order to determine the effective concentration range of drugs,MTT assay was used todetect the cell activity with the impact of different drug concentrations.The cells werepretreated before LPS stimulated with samples of different concentrations.Then Griess methods were used to detect the secretions of NO.