The Extraction, Separation And Purification Of EGCG3"Me

EGCG3"Me and EGCG4" Me,two O-Methylated EGCG derivatives,have shown significant efficacy on an anti-allergic action,especially on anti-pollen-allergic action.Therefore, the related research has attracted great attention in Japan,and it is also well acknowledged by world industries of medical and health products that it is highly possible for O-Methylated EGCG to be developed into a single variety and,consequently,to be a new growth focus in the Catechin economy,which will surely bring about a huge amount of wealth to the society.In order to fully explore the rich functional tea variety resources,increase the comprehensive value of tea resources and enrich the research on O-Mythelated EGCG in China,this study has applied HPLC,HPTLC polyamide,and HCCC have led to the extraction,separation and purification of EGCG3"Me.The research results are discussed as follows.1,The orthogonal experiment analysis leads us to the best technological condition for z EGCG3"Me extraction:70%ethanol,reflux extraction for 60min,temperature 60℃,and material and solvent ratio 1:10.2,Separation and purification of EGCG3"Me by of polyamide.a.preliminary separation of EGCG3"Me by way of polyamide.Crude polyamide is used to separate the EGCG3"Mepreliminarily,by means of gradient elution.Although there is no target compound in water washed liquid and 20%ethanol eluent,this method can remove part of the impurity and help the separation of target compound.Then,large amount of target compound is found in the eluent of 40%rthanol and 60%ethanol,while small amount exists in the 80%of ethanol eluent,indicating the fact that the 60%ethanol could elute most of the target compound.b.Further separation and purification of EGCG3"Me by polyamide.A further separation and purification of EGCG3"Me is realized by polyarnide with 100-120 mesh,and a high purity(40%)of EGCG3"Me is achieved.This method is of good repeatability and suitable for further extraction and separation.c.Purification of EGCG3"Me by HSCCC.When HSCCC is applied in the purifying of EGCG3"Me,the purity could reach 90%above. In the preparation experiment of countercurrent chromatography,the injection volume and the flow rate of mobile phase have great influence on the effect of separation.After the repeated experiment we get the best tevhnological condition:sample of injection volume 20mg solves in 30ml solvent,at a flow rate of 1.5ml/min.