Study On The Quality Standard Of Mongolian Medicine Cynanchum Thesioides

Cynanchum thesioides(Freyn)K.Schum is a characteristic Mongolian medicine named "Temo-Huhu",which belongs to the Asclepiadaceae Cynanchum plants.It is one of the basic ingredients of traditional Mongolian medicine prescription "indari-4 decoction" and "duguemonon" for treating intestinal hot dysentery and diarrhea.The appendix of the 1990 edition of the "Chinese Pharmacopoeia" was recorded the seeds of Cynanchum thesioides(Freyn)K.Schum."Drug Standards of the Ministry of Health of the People’s Republic of China(Mongolian Medicine Volume)" was contained Cynanchum thesioides(Freyn)K.Schum.for medicinal purposes.At present,the quality standard of Cynanchum thesioides(Freyn)K.Schum.is extremely imperfect.Only the characteristics and microscopic identification were included,without TLC identification,moisture,ash,extracts,and content determination item.Aim: To analyze the main components of Cynanchum thesioides(Freyn)K.Schum.,extract,isolate,purify and identify the component from the Cynanchum thesioides(Freyn)K.Schum.,and study the pharmacological activity of the components,the index component was selected,and the quality standard of Cynanchum thesioides(Freyn)K.Schum.was established.Methods: With help of UPLC/Q-TOF-MS/MS,the main component was determined in Cynanchum thesioides(Freyn)K.Schum.,which is named Thesioideoside,extracted and purified by diacolation,extraction,silica gel column chromatography,HPLC preparation,and recrystallization,identified by UV,Proton spectrum,Carbon spectrum,and Mass Spectrometry.Applying LPS to human normal colon epithelial cells(NCM460)to observe the effect of LPS on the proliferation of NCM460,then Thesioideoside was given to intervene in order to observe the effect of Thesioideoside on the proliferation of NCM460 cells,growth by MTT method,expression and release status of inflammatory factors in NCM460 cells by kit test,and regulatory effect of index component on inflammatory factors IL-8 and TNF-α.According to the Chinese Pharmacopoeia,the quality standard of Cynanchum thesioides(Freyn)K.Schum.was established,including TLC,moisture,ash content,alcohol-soluble extract,and Thesioideoside content by HPLC.Results: 1.By UPLC/Q-TOF-MS/MS analysis,Thesioideoside is main component in Cynanchum thesioides(Freyn)K.Schum.,which is obtained by diacolation(95% ethanol and 50% ethanol)separated from the n-butanol extraction,and identified by UV,Proton spectrum,Carbon spectrum,and Mass Spectrometry.2.0.01mg/m L and 0.02mg/m L of Thesioideoside have a significant protective effect on the growth inhibition of NCM460 cells by LPS,and can significantly reduce the levels of inflammatory factors IL-8 and TNF-α.3.The quality standard of Cynanchum thesioides(Freyn)K.Schum.was established.1)The eatablishment of TLC identification.2)Set the limit of its check items,the water not exceed 11.5%,the total ash content not exceeds 7.5%,and alcohol extract not less than 16.8%.3)The eatablishment of content determination of Cynanchum thesioides(Freyn)K.Schum.,Thesioideoside not less than 0.05% in dry medicine.Conclusion: 1.Thesioideoside,obtained from Cynanchum thesioides(Freyn)K.Schum.,is the main component by UPLC/Q-TOF-MS/MS,which can repair of LPS-induced NCM460 cell damage,and has certain pharmacological effect on diarrhea,so Thesioideoside can be used as an index component for establishing a quality standard for Cynanchum thesioides(Freyn)K.Schum..2.With Thesioideoside as index,the quality standard of Cynanchum thesioides(Freyn)K.Schum.was studied to form the "Draft quality Standard of Cynanchum thesioides(Freyn)K.Schum.",which is further improved and perfected the quality standard of Cynanchum thesioides(Freyn)K.Schum..