Extraction Of Alkaloids From Japamese Metaplexis Pericarp And Evaluation Of Their Bioactivity On Tca8113 Cells

ObjectiveJapamese metaplexis pericarp,containing many bio-functional compunds,is the fruit shell of Metaplexis japonica(Thunb.)Makino.The purpose of this experiment is to verify the inhibitory effect of the alkaloid from japamese metaplexis pericarp on Tca8113 cells through optimizing the extraction process of alkaloids from japamese metaplexis pericarp,studying the antioxidant activity of the crude alkaloids extractive,isolating the alkaloids as well as studying the structure characterization of alkaloids.MethodsTaking the japamese metaplexis pericarp as the research subject,based on the single factor experiment,the response surface method was used to optimize the extraction process of the alkaloids from japamese metaplexis pericarp.Single alkaloid was obtained by further isolating silica gel column chromatography.The polarimetry,NMR and MS was used to identify the structure of alkaloids.The scavenging ability of the alkaloids from japamese metaplexis pericarp to DPPH·,O-2·and·OH was evaluated by colorimetric test,and its in vitro antioxidant activity was evaluated.The activity of superoxide dismutase(SOD)in cells was measured.The inhibitory effect of the alkaloids on the proliferation of Tca8113 cells was evaluated by MTT assay,Hoechst 33342 staining.ResultsThe parameters of optimum extraction process of alkaloids are as follow:the extraction time is 40 min,the volume fraction of methanol is 88%,the extraction temperature is 20 ℃ and the ratio of liquid to material is 30∶1 m L/g.Under the best condition,the extraction amount of alkaloids is 230 mg/kg.Three alkaloids,secu’amamine D,securinine and securitinine,were isolated from japamese metaplexis pericarp.Colorimetric test shows highest scavenging capacity of alkaloids against DPPH·,O-2·and·OH is 87.43%,44.69% and 29.25%.The SOD experiment showed that the activity of SOD in Tca8113 cells was enhanced.MTT method showed that three alkaloids had significant inhibitory effect on Tca8113 cells.The results of Hoechst 33342 showed that all three alkaloids could cause the cell nucleus of Tca8113 cells to shrink and fragment.ConclusionsThe alkaloids from japamese metaplexis pericarp have certain antioxidant activity.Three alkaloids,secu’amamine D,securinine and securitinine,were isolated from japamese metaplexis pericarp.Three alkaloids can inhibit the proliferation of Tca8113 cells.