Separation，Purification And Antioxidant Acitivity Of Polyphenol In Black Wheat

Black-grain wheat belongs to hexaploid wheat. The difference betweenblack-grain wheat and common wheat appears in kernel pericarp and aleurone layercolor. Black-grain wheat is rich in natural melanin. It is a precious germplasmresource. compare to common wheat，The black-grain wheat has a higher content ofprotein, amino acid, element benefit to human body and antioxidant components.Soblack-grain wheat has very high application value and development prospects.In this paper，gallic acid was used as the standard and the phenols content ofblack-grain wheat were detected under the method of the Folin-Ciocaltu. analysisthe surface response to The optimal processing of ultrasonic extraction ofpolyphenols of black-grain wheat are analysised under the method of the surfaceresponse.the best condition of polyphenols purification of the macroporous resinAB-8to black-grain wheat were explored; the antioxidant properties were studiedby the following six system: total antioxidant ability, reductive abilitydetermination, linoleic acid system antioxidant ability, hydroxyl radical scavengingeffect, superoxide radical [O2ˉ] scavenging effect and DPPH clearance; use tomake structure preliminary appraisal of polyphenols in bran contain was detectedunder the method of infrared tandem mass spectrometry (ms) technology. The mainresearch result:(1) The research of polyphenol content determination of black-grain wheatThe best condition of using gallic acid as the standard and the FoLin-Ciocalteumethod to determine the content of bran polyphenol of black grain wheat:0.8mol/Lof FC reagent,10%of Na2CO3, reaction temperature35℃, warm bath after120min were measured in740nm in absorbency, gallic acid content determinationsystem in concentration range for2~10μ g/mL, and absorb light value was goodlinear relationship. Average recovery rate for determining methods is97.62%, RSD3.33%.(2) Black-grain wheat of polyphenols extraction process researchUse of ultrasonic wave extraction method and the response surface method tooptimize black wheat bran polyphenols extraction technology. The results show thatthe method is simple and easy for operation, character of higher yield. At the sametime, the regression equation can have better prediction of the relationship of testfactors and polyphenols yield. The optimum technology parameter for ethanolconcentration is58.12%, the extraction time of36.99min, material liquid than for1:21.97, extraction temperature for50℃, ultrasonic power for60W. The highest polyphenols rate occurred at3.545mg/g.(3) Purification process of polyphenols in black-grain wheatAnalysis the purification effect of black-grain wheat through the adsorptionperformance, elution ability examines and etc. It’s confirmed that the type AB-8macroporous adsorptive resin use as the material of polyphenols purification of theblackg-grain wheat bran through the static adsorption and analytical experiment.Through the dynamic adsorption test for AB-8type macroporous resin on thesample and elution factors of optimization, determine that the best technology is themost appropriate sample concentration of AB-8type macroporous resin purificationblack grain wheat bran of polyphenol, sample on speed, solvent elution volumefraction, elution speed were1.5mg/mL,2BV/h,60%ethanol,2BV/h. With thehighest rate of94.90%.(4) Preliminary identification of polyphenols structure of Black-grain wheatBy using Fourier infrared spectrometer experiment scanning black-grain wheatpolyphenols extract samples and compare with literature material, phenolic featureis apparent, and the information of tannins shows in the map. With reference to theliterature material for ESI-MS analysis, black-grain wheat bran contains theflavonoids polyphenols, anthraquinones polyphenol and flavanols compounds.(5) Research of vitro polyphenols antioxidant activity of Black-grain wheatUsing Vc as reference, and the following six methods to determine theantioxidant activity of the sample of bran polyphenols extract from black-grainwheat before and after purification: total antioxidant ability, reductive abilitydetermination, linoleic acid system antioxidant ability, hydroxyl radical scavengingeffect, superoxide radical [O2ˉ·] scavenging effect and DPPH clearancedetermination. We can conclude from the experiment results, black-grain wheatpolyphenol has strong antioxidant capability, and shows a quantity-efficiencyrelation with concentration. Antioxidant activity of purified sample is obviouslybetter than the sample before purification.