Anti-Tumor Activity Composition Screening Of Rumex Patientia L.Root

Objective:Rumex Patientia L.is a member of the Polygonaceae Rumex plant,which is one of the sources of Rumex madaio.It is mainly produced in northeast,north and northwestern provinces of China.Its medicinal parts are roots,with bitter,sour and cold taste.it has the effects of heat-clearing,detoxifying,blood-activating,hemostasis,purgation and desinsection.An innovative analytical approach based on the off-line two-dimensional reversed phase/reversed phase liquid chromatography?RPLC/RPLC?was applied for systematical separation of the active components and preparation of standard minor compounds in Rumex patientia L.Based on the research of chemical composition and pharmacological action,the anthraquinone glycoside in Rumex Patientia L.are extracted and enriched,and the chemical constituents of the plant are analyzed by qualitative analysis and chromatographic separation.The antitumor activity of the separated components was screened in vitro test,the active components were obtained and the antitumor activities in vitro were further studied,the IC₅₀ of each active component was calculated,and their antitumor effects were evaluated and compared.Methods:Used Unitary C18 and a kind of new reverse stationary phase developed by our seminor,Two-dimensional liquid chromatography analysis was carried out to seprate and analyze the chemical constituents of the ethyl acetate layer in Rumex patientia L.By system solvent extraction,solid phase extraction and high pressure preparation liquid chromatography,the anthraquinone glycoside in Rumex Patientia L.were extracted and separation with high efficiency and high automation.UHPLC-MS was used for qualitative analysis and identification of the anthraquinone glycoside in Rumex Patientia L.CCK-8method was Detected in;final analysis use UHPLC-MS technology to obtain molecular ion peak and fragment ions peaks to identify the compounds in fractions.Results:Based on the experiment setup and result,it was concluded that off-line 2D-LC could be effective method in future to separate and analyze the chemical constituents of the ethyl acetate layer in Rumex patientia L.31 kind of anthraquinone glycoside were obtained by chromatographic separation.21 of the anthraquinone glycoside compounds of Rumex Patientia L.were identified by UHPLC-MS.After screening and analysis of cell activity,4components with antitumor activity were selected,including BTS-F1-19,BTS-F1-20,BTS-F1-21 and BTS-F2-8.Finally,it was found that BTS-F1-19 has stable anti-tumor activity,and its inhibition rate to tumor cells was better after determining the anti-tumor activity in-depth.Conclusion:The results of this study provide reliable basis for the separation and analysis of chemical constituents and credible data of the anti-tumor mechanism of Rumex Patientia L..