The Study Of Purification And Identification Of Total Phenolic Acid In Ilex Asprella

Objective:Nowadays,the compounds study research of Ilex Asprella is mainly about saponin compounds,the phenolic acid was reported but not to study indeeply.Therefore,ingredients like this paper to phenolic acid as the research,to established a quantit-ative determination method of total phenolic acid and total saponins in Ilex Asprella by ultraviolet spectrophotometric.Then determined 20 batches of asprella herbs.To optimize the extraction condition for phenolic acid in Ilex Asprella by response surface methodology and the refining technology by macroporous resin.At last phenolic acid were isolated and identified.Methods:1 Study on the extraction technology of total phenolic acid in Ilex Asprella1.1 Refluxed extract the total phenolic acid in Ilex Asprella by 70%ethanol and determined the absorbance at 312nm.1.2 Based on the single factor experiment to optimized the ethanol concentration,extraction time and solvent dosage by used response surfa-ce method.2 The process of macroporous resin purification of total phenolic acid in Ilex AsprellaBased on the content of total phenolic acid in Ilex asprella,compar-ison of different types of resin on theseparation and purification capac-ity of total phenolic acid in Ilex asprella.By investigating the ethanolconcentration,elution solvent dosage and elution velocity,to establish purification conditions of total phenolic acid in Ilex Asprella with macroporous adsorption resin.3 Isolation and identification of phenolic acids in Ilex AsprellaIsolation to applicate ODS,HPLC,preHPLC chromatographic technique.Identificated compound apply to NMR,MS.Results:1 Study on the extraction technology of total phenolic acid in Ilex Asprella1.1 In the range of 2.12?10.6 u g/mL,there is a good linear relations-hip(r=0.9995);The average recovery is 100.7%,RSD is 2.08%.The contents of twenty batches samples between 0.1%and 0.3%.1.2 The optinal condition was as follows mass concentration of ethanol was54%,extraction time was 88min and the amount of solvent was33mL.To compared the effect of total phenolic acid extraction rate on sequential,the mass concentration of ethanol(A),the amount of solvent(C)>extraction time(B).2 The process of macroporous resin purification of total phenolic acid in Ilex AsprellaX-5 macroporous resin' s adsorption and disadsorption ability was bet-ter,the best technological conditions were 40%ethanol as elution agent,volume of elution agent was 2 BV,concentration of sample was 1.57mg/mL and the flow eluting velocity was 2.OmL/min.In these condition,effectively reduced the amount of solids,increased the overall amount of acid,and the phenolic acid content was 6.31%,purity reached 445.6%,RSD was 0.6%.3 Isolation and identification of phenolic acids in Ilex Asprella There were 5 phenolic compounds isolated from Ilex Asprella,the compounds were identified as caffeic acid,methyl caffeate,3-caffeoylquinic acid,4-caffeoylquinic acid,3,5-di-O-caffeoylquinic acid.The caffeic acid have been reporyed,the other 4 compounds were isolated from this plant for the first time.Conclusions:1 Study on the extraction technology of total phenolic acid in Ilex Asprella1.1 The method of ultraviolet spectrophotometric is simple,sensitive,accurate and reliable,can be used for determination of total phenolic acid in Ilex Asprella.1.2 Optimization of extraction technology can effectively reduce bli-ndness experiments by using response surface methodology.Provide a theo-reticcal basis for the further development use of Ilex Asprella.2 The process of macroporous resin purification of total phenolic acid in Ilex AsprellaX-5 macroporous resin have good ability to enrich and purify total phenollic acid in Ilex Asprella,the process was simple,high degree of continuous production,suitable for total phenolic acid separation and purification.3 Isolation and identification of phenolic acids in Ilex AsprellaThe phenolic compounds which isolated from Ilex Asprella not only enrich the studies on chemical consitituents of this plant,also provides the material basis for the Ilex Asprella formulate quality standard.