The Research On The Chemical Ingredient And Tissue Culture Of Ardisia Pusilla A. DC.

Ardisia pusilla A.DC.has a long history for medicine use,in which the efficacious part is Ardipusilloside that mainly existed in the stem and root of Ardisia pusilla A.DC.Recently research had showed that Ardipusilloside have taken on prominent efficacy on raising immunity and anti-tumour,thus Ardipusilloside had an enormous prospect for developing anti-tumour drug,therefore,it was actively prepared for anti-cancer drug development. Ardipusillosideâ… ,the main ingredient in Ardipusilloside,thus can be applied to measuring the quantity of Ardipusilloside;But Ardisia pusilla A.DC.are still living in the wild,the cultivation and research focusing on its growth are still blanked.The research on chemical ingredient of Ardisia pusilla A.DC.consists of the extraction and purification of Ardipusilloside,the separation Ardipusillosideâ… from Ardipusilloside.The optimum extraction method was selected by using of orthogonal L₉(3⁴)experiment,fixing Ardipusillosideâ… contents and dry extract rate as index,The optimal technology was elected as 75%ethanol,using 8 volume times solvent by 4 hours on the first extraction,6 volume times solvent by 3 hours on the second extraction.Ardipusilloside was purified through the resin in order to reach to 50%according to the administrative regulations pertain to theâ…¤Chinese medicine declaration.In addition,Ardipusillosideâ… was separated from the Ardipusilloside through the silica gel column and preparation HPLC sequently,which was indentified.the purity was reached to 99%,;HPLC—ELSD determination of Ardipusillosideâ… was established,which is Chromatogram condition:PhenomenexC₁₈(4.6×250mm,5u),Flow phrase:methanol:water(75:25),Volume flow:1.0 mL·min^-1,Tube temperature:73.8℃,gas flow:2.0 L·min^-1.In order to supplement the blank in cultivation and growth characteristics of Ardisia pusilla A.DC.and establish the plant resources,the tissue culture was applied to cultivate Ardisia pusilla A.DC.,which included sterilization of explants,the induction of callus and establishment of cell suspension culture system of Ardisia pusilla A.DC.,the examination on the process from the seed germination to mature plant,the discovery of "multi-embryo" phenomenon.The best induction of callus medium was discovered,which is MS culture Adding 3 mg/L 6-BA and 0.3mg/L2,4-D,the inducing rate could run up to 100%.The optimum medium suitable for Ardisia pusilla A.DC.was examined,which is MS culture adding 5.5g/L agar.