Survey On The Resource Of The Effective Constituents From The Vitex And The Purification Of The Effective Fractions Against Tumor From Fructus Viticis Negundo

Vitex negundo L.grows widely throughout the south of Yangtze River.The fruits,roots,stems,leaves,and other parts of this plant can be used as a medicine with analgesical,anti-inflammatory,anti-bacterial, antioxidant and anti-tumor activities,meanwhile it can regulate the hormone secretion.In the early investigations,the studies of the chemical constituents from the effective anti-tumor parts of Vitex negundo L..led to the isolations of a series of phenyldihydronaphthalene -type lignans,particularily VBE-1 and VBE-2.Both of them showed strong inhibitory in vitro and in vivo activities against the growth of human breast cancer,ovarian cancer,human hepatocellular carcinoma and human colon cancer cells.A HPLC method to determine the content was established by the mean of study for the VBE-1 and the VBE-2 at the first time.The experimental conditions as follows:Kromasil ODS-1 column(250mm×4.6mm,5μm) was used as the chromatogram column,the mobile phase was acetonitrile -0.1%acetic acid aqueous solution gradient elution(0 min,15:85;10 min,15:85;40 min,28:72);detection wavelength was at 255 nm,the flow rate was 1.0 mL/min,and the column temperature was at 25℃.A good linear correlation was obtained when concentrations of VBE-1 and VBE-2 ranged from 0.430～6.45ug and 0.239～3.59ug separately.The investigation about the resource of this plant was studied for the purpose of finding sufficient and highly efficient anti-cancer drugs.The results showed that the content of VBE-1 and VBE-2 from Vitex negundo L.were higher than those in Vitexin;And in all parts of Viticis negundo L.,the content of the fruits was much higher than those of the roots, stems and leaves.Besides,the different environments of different areas also influenced the content of VBE-1 and VBE-2,according to the research the content was higher relatively in Guizhou province and the Shuangpai city from Hunan province.The subject aimed at gathering the phenyldihydronaphthalene-type lignans from fruits of Vitex Negundo which have obvious anti-tumor activity.The process of this experiment focused on extraction and purification by polyamide column chromatography.Finally the process was optimized as follows:The fructus of Vitex Negundo was extracted with 40%ethanol twice(1.5h and 1.0h,respectively) and the volume of solvent was respectively 8,7 times of the medicine amount.The extract was concentrated to remove the ethanol and absorbed in polyamide (30-60 mesh) with 0.4 times of the medicine volume,which was subjected to column chromatography over polyamide(30-60 mesh) with 0.6 times of the medicine volume,using water(2BV) and 0.5%ammonia water(4BV) as gradient elution.The eluent of 0.5%ammonia water was absorbed in polyamide(30-60 mesh) with 0.4 times of the medicine volume,which was subjected to column chromatography over polyamide (30-60 mesh) with 0.6 times of the medicine volume,using H2O-EtOH as gradient elution(H2O:2BV,40%EtOH:4BV).The ethanol of 40% EtOH eluent was removed under pressure and extracted twice with ethyl acetate(v:v 1:1).The ethyl acetate layer was evaporated in vacuum at 60℃to afford the effective portion with VBE-1 and VBE-2 accounting for 50%.