Study On The Extraction Process And Properties Of Triterpenoid Acid

Triterpenoid acid as a pentacyclic triterpene compound because of it,s antibacterial,antiviral,antitumor,antioxidant,hepatoprotective,lipid-lowering,hypoglycemic,increase the number of white blood cells,enhance immunity and other important physiological functions and become the research focus of today's society.In this study,the detection methods,extraction process,purification process and in vitro antioxidant activity of the triterpenoid acid were studied to provide technical support for the further development and utilization of the compounds in the raw materials of agricultural products.The main research contents and results are as follows:In this study,a simple and sensitive HPLC-UV method was firstly established for simultaneous detection of betulinic acid,oleanolic acid and ursolic acid.A Kromasil C₁₈column?5?m,4.6×250 mm?was used for the separation at ambient temperature with methanol and water?employed phosphoric acid adjusting PH value to 3.0?as the mobile phase at a flow rate of 0.6 mL/min,and the detection wave length was set at 210 nm.All calibration curves showed good linear regression?R²?0.9998?within the linear ranges of2¹00?g/mL.The limitation of detection and quantification were 0.21⁰.26?g/mL and0.71⁰.87?g/mL,respectively.The precision,stability,repeatability and accuracy of the analytical method meet the requirements of experiment.The established analytical method was then applied to a simultaneous detection of the three kinds of triterpenic acids from different types and different varieties of raw materials and finally the Fuping jujube was chosen as raw materials.In this study,we compared the effect of immersion extraction technology and ultrasonic assisted extraction technology for the extraction yield of betulinic acid,oleanolic acid and ursolic acid.Finally,the ultrasonic assisted extraction method was chosen as the final extraction method for the three kinds of triterpenoid acid in Fuping jujube.Three parallel experiments carried out under the optimal extraction conditions(ethanol concentration was 95%,extraction temperature was 52?,extraction time was 17min,liquid/solid ratio was 23:1 mL.g^-1,repeated extraction two times)resulted in a total extraction yield of three kinds of triterpenoid acid of 670.15±30.12?g/g.In this study,enrichment and purification of triterpenoid acid from Fuping jujube extracts were studied using different resins.The static tests indicated that AB-8macroporous resin was appropriate.To optimize the separation process,dynamic adsorption and desorption tests were carried out.The optimal adsorption parameters were initial concentrations in sample solution of 65.37,27.17 and 50.92?g/mL for betulinic acid,oleanolic acid and ursolic acid,respectively,do not adjust pH,flow rate of 2 BV/h,sample loading amount of 3 BV.The optimal desorption parameters were deionized water and 40%ethanol each 3 BV,then 95%ethanol of 3 BV,do not adjust pH,flow rate of 2BV/h,collect eluent of 1².5 BV.After one run treatment with AB-8 macroporous resin,the content of triterpenoid acid in the product increased from 0.55±0.02%to 11.69±0.54%,and the recovery yield was 76.32±3.79%.The results showed that AB-8macroporous resin revealed a good ability to enrichment and purification triterpenoid acid from Fuping jujube extracts.In this study,the antioxidant activities of betulinic acid,oleanolic acid,ursolic acid standard solutions and Fuping jujube purified solution were evaluated.The results showed that betulinic acid,oleanolic acid,ursolic acid,three kinds of triterpenoid acid mixed standard solution,Fuping jujube purified solution and reference solutions ascorbic acid,BHT had high potent scavenging activity toward.OH,with the IC₅₀ values were 0.172?0.228?0.217?0.213?0.137?0.199 and 0.184 mg/mL,respectively.The IC₅₀ values of scavenging activity toward O₂^-.were 0.073?0.070?0.082?0.079?0.065?0.158 and 0.067mg/mL,respectively.The IC₅₀ values of scavenging activity toward DPPH radical were1.288?0.343?0.754?0.473?0.042?0.029 and 0.082 mg/mL.The IC₅₀ values of scavenging activity toward ABTS^+.were 0.862?0.251?0.466?0.792?0.077?0.052 and 0.026 mg/mL.The reducing ability increases with the increase of sample concentration.Betulinic acid,oleanolic acid,ursolic acid,three kinds of triterpenoid acid mixed standard solution,Fuping jujube purified solution and reference solutions EDTA had high potent scavenging activity toward Fe²⁺,with the IC₅₀ values were 0.618?0.665?0.572?0.482?0.984 and 0.021mg/mL.Indicated betulinic acid,oleanolic acid,ursolic acid standard solutions and Fuping jujube purified solution all had high antioxidant activities.