Study On Extraction And Purification Of Dioscin From Trigonella Foenum-graecum

The ultra-high pressure extraction technology,or UHPP for short,in this experiment,was used to extract dioscin from Trigonella foenum-graecum.High-purity dioscin was prepared by preparative high performance liquid chromatography（P-HPLC）on the basis of separating and purifying dioscin with foam separation,the products were characterized by FT-IR,HRMS and so on.The specific contents of the study are as follows:Firstly,a new extraction process,named ultrahigh pressure extraction technology,was used to extract dioscin from trigonella foenum-graecum.The process conditions were optimized by response surface methodology（RSM）and the effects of ultrasonic extraction and ultrasonic extraction conbined with complex enzyme on the yield of dioscin were studied.The optimum solid-liquid ratio,ultra-high pressure holding time,holding pressure and ethanol concentration were 1:30,563 s,360 MPa,71%,the yield of dioscin was up to 5.02%.And the yield of dioscin were 2.27%and 3.36%by ultrasonic extraction and ultrasonic extraction conbined with complex enzyme,respectively.Compared with two traditional extraction methods,ultra-high pressure extraction method was considered to be effective for extracting dioscin from trigonella foenum-graecum.Secondly,a new method for purification of dioscin from trigonella foenum-graecum by foam fractionation was established based on the extraction of dioscin by ultrahigh pressure extraction technology.Based on the single factors experiments,the effects of solvent concentration,temperature,volume of liquid and air flow rate on the enrichment ratio and recovery of the dioscin from trigonella foenum-graecum were optimized by RSM.At the same time,its antioxidant activity in vitro was also studied.The results indicated that 0.03 mg/mL,26℃,450 mL/min and1:162 were the optimum separation parameters for solvent concentration,temperature,volume of liquid and air flow rate,respectively.Under this condition,the enrichment ratio and recovery of the dioscin were 88.23%and 1.25.The antioxidant activity study showed that abilities of dioscin in trigonella foenum-graecum has strong antioxidant activity.Thirdly,the concentrated foam was further purified by P-HPLC after the foam separation,Chromatographic separation was performed on a Hedera ODS-2 column（20×250 mm,10μm）with mobile phase of acetonitrile-water solution with gradient elution at the flow rate of 10 mL/min,the injection volume was 5 mL.Finally,the products obtained were characterized by HPLC,FT-IR,HRMS,¹H-NMR,¹³C-NMR.