Study On Extraction,Purification And Antioxidant Activity Of Polyphenols And Polysaccharides From Cold Pressed Oil Cake Of Zanthoxylum Armatum DC. Prodr.

In order to investigate the application value of polyphenols and polysaccharides in cake,the Zanthoxylum armatum DC.Prodr.and cold pressed oil cake were adopted as the main materias to conduct a comparative study on the content and antioxidant activity in vitro of polyphenol and polysaccharides.Thus,theoretical reference for the evaluation of cold pressing technology and the recovery and reuse of the cake could be provided..The main results are as follows:（1）On the basis of the single factor test,the optimum extraction of PPOC conditions were studied by orthogonal test.The optimum extraction conditions were as follows:material liquid ratio 1:25,microwave treatment time 360 seconds,ethanol concentration of 50.00%,microwave power of 400 W,extraction temperature of 60 ℃.Under these conditions,the yield of PPOC was 6.83±0.09 mg/g,and the yield of PPOP was 15.45±0.09 mg/g.（2）The purification effect of three kinds of macroporous resins with different polarity on PPOC was compared.The results indicated that AB-8 macroporous resin owned a better purification effect on polyphenols.The optimum process conditions of AB-8 for purifying PPOC were:the pH value of the sample solution was 4,the concentration of polyphenols was 0.53 mg/mL,the loading flow rate was 1.0 mL/min,and the sample was eluted with 60.00%ethanol at a flow rate of 1.0 mL/min.The purity of PPOC increased from 35.70%to 76.30%after being purified by macroporous resin,meanwhile the purity of PPOP increased from 29.52%to 70.20%.（3）On the basis of single factor test,the optimum extraction of PSOC conditions were studied by response surface methodology.The optimum extraction process conditions were identified as follows:microwave power 505 W,extraction time of 16 min,extraction temperature of 85 ℃,liquid to material ratio 44:1,the yield of PSOC was 4.96 ± 0.07 mg/g.This model could be applied to predict the yield of PSOC.（4）The protein in PSOC was precipitated by TCA（trichloroacetic acid）method.The optimum conditions were as follows:the dosageof TCA was 6.00%,oscillated at constant temperature for 15 minutes,processed for once.After treated by TCA,polyamide column chromatography was applied for the further decolorization and deproteinization:the loading volume was 50 mL,the loading concentration of the sample was 4.00 mg/mL,the adsorption equilibrium time was 25 min,the elution rate was 1.50 mL/min.After the decolorization and deproteinization,the deproteinization rate of PSOC was 90.59%,the decolorization rate was up to 94.23%,and the retention rate of polysaccharide was 61.94%.After the two-step purification above,the purity of PSOC could be increased from 48.52%to 87.93%.The purity of PSOP was increased from 52.17%to 89.25%.（5）The IR analysis indicated that PSOC was mainly composed of a-configuration pyranose.The monosaccharide composition of PSOC and PSOP were determined by HPLC-PMP derivatization method.Monosaccharide composition of PSOC were rhamnose,galactose acid,glucose,galactose,xylose,and could possibly contain glucuronic acid.The results revealed that PSOP could keep most of the monosaccharide in PSOP.（6）The antioxidant activity of polyphenols and polysaccharides in vitro was analyzed by typical antioxidant systems.The results indicated that the antioxidant activity of PPOC was lower than that of PPOP.The reducibility and O₂^-· scavenging ability of PPOC was superior to that of BHT.The antioxidant activity of PSOC in vitro was lower than that of PSOP and VC.The IC50 values of BHT,PPOC,PPOP against ABTS · +were:0.005,0.052,0.063 mg/mL,IC50 values against DPPH.were:0.054,0.090,0.126 mg/mL,IC50 values against 02-· were:0.234,0.087,0.118 mg/mLIC50 values of VC,PSOC,PSOP against ABTS ·+ were:0.019,0.237,0.304 mg/mL,IC50 values against O₂^-· were:0.059,0.569,0.836 mg/mL.