Identification Of Antitumor Active Constituents Based On The Composition-activity Relationship Model Of Aconitum Carmichaelii-Liquorice

The object of the study is Radix Aconiti Carmichaeli and Licorice.Based on methods of HPLC as well as HPLC-MS/MS,a database of chemical components in aqueous extract of Radix Aconiti Carmichaeli and Licorice was established.Combining with the antineoplastic activity of 31 batches of aqueous extract from different sources collected in vitro cell research,we built up a model of composition-activity relationship and identified the potential lead active compounds in aqueous extract of Radix Aconiti Carmichaeli and Licorice by using method of MIV,which could provide the basis for the further study,such as compatibility of active compounds and development of new anticancer drugs.A method for analyzing the aqueous extract of Radix Aconiti Carmichaeli-Licorice was established by HPLC using Waters e2695 HPLC.Several factors have been studied,including chromatographic column,composition of mobile phase,elution program,determine wavelength,column temperature and flow rate.Chromatographic conditions are determined as follows:chromatographic column of Waters Symmetry C₁₈,mobile phase of 5mM ammonium acetate?0.1%glacial acetic acid??A phase?-acetonitrile?B phase?,gradient elution,determine wavelength of 254nm,flow rate of0.4m L/min,column temperature of 30?.The method was used in the determination of31 batches of aqueous extract from different sources.30 characteristic common peaks were quantified by inter-standard quantitative method using lappaconitine as the internal standard substance.Both of the relative retention time and relative peak area were analyzed to make the study of methodological observation.The result which showed all the RSDs?5%proved that the established method was accurate,reproducible and stable in 24h.Based on the established HPLC method for analyzing the aqueous extract of Radix Aconiti Carmichaeli-Licorice,the corresponding chemical component,compound structure and source from which medicine of 30 characteristic common peaks were studied by combined utilization of the MS in positive ion mode.According to the information of fragment ions collected in both HPLC-MS and HPLC-MS/MS as well as numerous relevant documents,29 components were identified successfully among30 characteristic common peaks.18 components were derived from Radix Aconiti Carmichaeli,which were all alkaloids,including diester diterpenoid alkaloids,monoester diterpenoid alkaloids and alkamine diterpenoid alkaloids.11 components were derived from Licorice,including flavonoid,triterpenoid saponins and so on.A PSO-SVR model of composition-activity relationship in Radix Aconiti Carmichaeli-Licorice was established,using relative peak area of 30 characteristic common peaks in 31 samples of different sources as independent variable and inhibition ratio of Hela cell multiplication for each sample as dependent variable.Sorting by the absolute mean impact value?MIV?of each component in the exact of Radix Aconiti Carmichaeli-Licorice,the top 15 components were identified as potential lead active compounds whose absolute MIV>0.01.Among the 15 components except for 1component which was not determined yet,11 components were derived from Radix Aconiti Carmichaeli and 3 components from Licorice.Most of the identified active compounds have been proved to be anti-cancer in the literature.