| Podophyllum is one of Berberidaceae Dysosma plants and has good antitumor activity.It's efficacy mainly comes from lignans.The targets of this study were to optimize the reflux extraction process of podophyllotoxin,research the isolation and purification technology of podophyllotoxin by microporous resin,analyse the extravts by HPLC-MS/MS.The main work was listed as folloes:1.Reflux extraction process was optimized by singer factor test,the Design of the CCD Response Surface method.The optimum extraction technology is as follows: ethanol concentration of 70% alcohol,ratio of liquid to solid of 10,mass ratio of liquorice and podophyllum of 11.5,extraction time of 3 hours and water-bath temperature of 100 degrees.As a result,the yield of podophyllotoxin is 62.62mg/g.2.The extract was purified by LSA40 resin which was choosed through static adsorption and desorption experiment.The adsorption equilibrium data was agreed with Freundlich isotherm well;The adsorption process of podophyllotoxin on macroporous resin LSA40 could be described well by pseudo-second order model.Fixed bed model could describe the adsorption kinetics well.The optimum isocratic isolation process is as follows: 3BV of extract with pouring velocity of 2BV/h,using 5BV 90% alcohol as eluting agent with velocity of 4BV/h,the content of podophyllotoxin rose from 0.81% to 2.85%,the recovery rate was 59.10%.The optimum gradient isolation process is as follows: first,the resin was eluted by 5BV of distilled water to wipe off impurities;second,the resin was eluted by 1BV of 10% alcohol,1BV of 30% alcohol,1BV of 50% alcohol and 5BV of 90% alcohol with velocity of 4BV/h,the content of podophyllotoxin was 2.96%,the recovery rate was 48.33%.3.HPLC-MS/MS method was used to determine the chemical structure of extract and we finally fond twelve kinds of lignans and two kinds of flavonoids.Besides,there was one kinds of isomer of dysosmarol identified. |
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