The Study On Extraction, Purification Of The Main Active Ingredients From Sweet Tea

With sweet tea as the research object,this paper stuides the decolorization condition of Sweet Tea sample solution,the extraction and purification process of rubusoside 、 total polyphenols and total flavonoids in Sweet Tea,the further purification of rubusoside and polyphenols in Sweet Tea,the component identification of rubusoside,the mental content in A5’、A6’、B4’was tested.The main conclusions were as follows:(1)Nine kinds of macroporous resin were selected by studying the decoloration rate and the retention rate of Rubusoside. The static and dynamic adsorption test of single factor experiment had been done to investigate the best decolorization conditions and regeneration conditions of the selected rein. The results showed that D296 macroporous resins was the best one. The optimal decolorization conditions were as follows: the p H was 6.5, the flow rate was 2.0 BV/h, concentration of sample 3.0 mg/m L(the concentration of polyphenols).The optimal regeneration conditions were: flushing the used macroporous resins with 3% Na OH and HCl solution until the effluent solution was nearly colorless.(2)Technology of meanwhile preparing three main products(rubusoside、total polyphenols and total flavonoids)from sweet tea was designed:First of all the sample solution of sweet tea was enriched by the SA-3 macroporous resin and the column flow fluid was collected.Then the saturated rubusoside enrichment column was eluted by low concentration of ethanol solution, the eluent and column flow fluid were combined and purified by macroporous resin HPD826.Then the enrichment column was eluted again by a high concentration of ethanol solution,eluent was dried by the rotary evaporator,and crude product of rubusoside was got.Polyamide resin was used for refining crude product,column flow was collected and concentrated by rotary evaporator、freeze-dried,then the high-quality product of rubusoside was got.Then the polyamide resin was eluted by 75% ethanol solution,column flow and concentrated by rotary evaporator 、 freeze-dried, and the product of total flavonoids from sweet tea was got.Besides,The optimal column conditions of SA-3 were as follows:Sweet Tea sample solution was used to get through the column with 3.0 BV/h、16 BV, collecting the flow fluid,Y2 was got.Flushing the SA-3 macroporous resins by 20% ethanol solution,7BV,collect the eluent then B2 was got.Combining Y2 and B2,B3 was got.Then flushing the SA-3 macroporous resins by 70% ethanol solution,collecting the eluent,A5 was got.The purity of A5’could reach about 45.7%,the yield was about 7.1%.Column condition of HPD826 resin were flow rate of 3 BV/h, 2 BV and flushing with 40% ethanol solution 3 BV as eluent.Eluent was collected and concentrated by rotary evaporator、freeze-dried.The purity and yield of the total polyphenols production were 60%、10%.Column condition of polyamide resin were flow rate of 2 BV/h,A5 sample solution volune of 2 BV,collecting the flow fluid then A6 was got.The purity and yield of A6’were 71%、3.4%.Then flushing the polyamide resin by 75% ethanol solution with flow rate of 2 BV/h,5 BV.collect the eluent then C1 was got.The purity of C1’could reach about 55.9%,the yield was about 0.81%.Methods of solvent extraction and recrystallization was used to further purifiy the A6’.Then A7’was got.The purity of A7’was 92%.HPLC、IR、LC-MS were used to prove that main composition in A7’was rubusoside.Methods of solvent extraction and Sephadex LH-20 resin was used to further purifiy B4’,then B11’was got.The purity of B11’was 73%.(4)AAS and AFS were used to determine the content of K、Na、Cu、Fe、Zn、Mn、Ca、Mg、Pb、Cd、Hg、As in the A5’、A6’、B4’,The results showed that those elements content in A5、A6 ’and B4 ’conform to the national relevant standards.