33 7 2017 7 Vol.33,No.7 Jul.2017 DOI:10.13652/j.issn.1003-5788.2017.07.033 Extractionofpolysaccharidefromgerminatedbrownricebyaqueous two-phasesystemandresearchofitsantioxidantactivity 1,2 2 1 2 LIU Xiao-fei 1,2 GONG Li-li 2 WANG Xin 1 MENG Qing-hong 1 MA Yong-qiang 2 ZHANG Yu 2 2 2 1 LU Shu-wen 1 (1., 150086 2., 150076) (1.Heilongjiang Academyof AgriculturalSciences PostdoctoralProgramme,Harbin,Heilongjiang150086,China 2.Key Laboratory for Food Scienceand Engineering,Harbin Universityof Commerce,Harbin,Heilongjiang150076,China) mined.results were showed as folows:the mass fraction of PEG6000was15.7%,themassfractionofammonium sulfatewas 14.8% andextractiontimewas40 min.theyieldofpolysaccharide substancefrom germinatedbrownricewas81.07% intheoptimum conditions.morever,theanti-oxidationofpolysaccharidesubstance fromgerminatedbrownricewerestudiedbymethodsofdpph,py-, rogalolautoxidationandphenanthroline.theresultsshowedthat, thepolysaccharideofbrownricehadstrongerantioxidantefectto, DPPH,ultraoxygenanionandhydroxylradicals. : Keywords:Germinatedbrownricepolysaccharideultrasonictwo- phaseextractionantioxidantactivity PEG6000 PEG600015.7% 14.8% 40min, 81.07%, DPPH, DPPH [1],, γ- [2-3], Abstract:Thepolysaccharidewasextractedbytheultrasonic-assisted, aqueoustwo-phasetechnologyfrom germinated brown rice.the [4] phaseseparationabilityofdiferentinpeg/(nh4) 2SO4system was studiedaccordingtothepolysaccharideyield.theyieldofpolysac- charidesubstanceofgerminatedbrownrice wasdeterminedbyor- thogonalexperiment.theoptimum processconditions weredeter- (LBH-Z15200), (12541189),,,, (1968 ),, [14-15] γ- E-mail:shuwenl@sina.com 2017 05 16, [5-6], [7] [8], PEG 3, 55.1% XingJianmin [9], [10],, /K2CO3 [11-13] [16-17] [18-19], 149
2017 7,, 1.3.4, (PEG)/ (1)PEG 20 ml,, 5.0 g, 4.0 g PEG2000 PEG4000, PEG6000 PEG8000 PEG10000,, 40min, 1 1.1 (2)PEG : 20 ml,, 5.0g,PEG6000 3.0,3.5,4.0,4.5, 1.2 5.0g,, 40 min :JY92-ⅡD,,, (3) : 20 ml, :SP-752PC, PEG6000 4.0g, 3.0,4.0,5.0, 6.0,7.0g,, 40 min :TGL-16G,,, HHS-12, 5.0g,PEG6000 4.0g,, :BS224S,, 1.3 1.3.1, (1)DPPH [23], (4), 2h, DPPH D, ( 30 A0 - ( Ai -Aj ) D = 100%, (4) 98% 24h) [20] A0 80 30 min, 55, (80) [19] 1.3.2 1.3.3 10.0g,, 1 10 (g/ml), 500 W 25min [22], 50,,3000r/min 20min,, 1000mL, (5) O2 - A 20mL, PEG, ΔA0 -ΔA,, 40 min A = 100%, (5) ΔA0,620nm, [21-22] K = Ca, (1) Cb R = Va, (2) Vb Y = RK (6) OH E 1+RK, ( 3) A -A E = 100%, (6) A -A K PEG/ E OH,% 150 R Y,% Ca,mg/mL Cb,mg/mL Va,mL Vb,mL (4) 20mL, 20,30,40,50,60min, 1.3.5 D DPPH,% A0 Ai A j (2) (O 2 - : [24], A O 2 -,% ΔA0 ΔA (3) ( OH) : [25], A A A 2 2.1 1
33 7 1 Figure1 Thestandardcurveofglucose :y=0.012x-0.0035,r 2 =0.9896, 2.2,, PEG 2.2.1 PEG 15.7% (65.03%), PEG,, 2~4 2~4,,PEG, R, K PEG, PEG, [8], K,PEG6000,,PEG6000, 75.27% 2.2.2 PEG 5~7 5~7, PEG, R, K, [28] 2 K PEG Figure2 ThecorrelationofpartitioncoeficientK and PEG molecularweight 5 K PEG Figure5 ThecorrelationofpartitioncoeficientK and PEGconcentration 3 PEG Figure3 ThecorrelationofvolumeratioandPEG molecularweight 6 PEG Figure6 ThecorrelationofvolumeratioandPEG concentration 4 PEG Figure4 Thecorrelationofpolysaccharidesyieldand PEG molecularweight 7 PEG Figure7 Thecorrelationofpolysaccharidesyieldand PEGconcentration 151
2017 7 2.2.3 8~10 8~10,,2 14.8%,, 20.6%, 2, 11 K 14.8% (73.25%) Figure11 ThecorrelationofpartitioncoeficientK and [27], PEG, 14.8% extractiontime 2.2.4 11~13 11~13,,, 40min, 12 Figure12 Thecorrelationofvolumeratioand extractiontime 8 K Figure8 ThecorrelationofpartitioncoeficientK and concentrationofammoniumsulfate 40min 9 2.3 Figure9 Thecorrelationofvolumeratioandconcentration, ofammoniumsulfate PEG PEG, 1 2 10 2.4 Figure10 Thecorrelationofpolysaccharidesyieldand concentrationofammoniumsulfate 152 13 Figure13 Thecorrelationofpolysaccharidesyieldand extractiontime,,,, 2, : PEG > >PEG > A2 B3 C2 D2, PEG6000 PEG6000 15.7% 14.8% 40min,, 3,, 81.07% 2.4.1 DPPH DPPH 14
33 7 1 Table1 Orthogonalarraydesign APEG BPEG C D /% /% /min 1 4000 14.3 11.5 30 2 6000 15.7 14.8 40 3 8000 17.2 17.8 50 15 2 Figure15 Scavengingefectofpolysaccharidefrom germi- Table2 Experimentaldesignandresultsofpolysaccharide natedbrownricebyo 2 - method fromgerminatedbrownrice 2.4.3 A B C D /% 1 1 1 1 1 63.86 2 1 2 2 2 69.30 3 1 3 3 3 63.50 4 2 1 2 3 77.03 5 2 2 3 1 76.25 6 2 3 1 2 74.63 7 3 1 3 2 65.76 8 3 2 1 3 68.53 9 3 3 2 1 67.05 k1 0.642 0.677 0.675 0.677 k2 0.726 0.676 0.680 0.679 k3 0.664 0.679 0.677 0.676 ( OH) 16 16,, 0.5 mg/ml,, 53%, [29] R 0.084 0.002 0.005 0.002 16 Figure16 Scavengingefectofpolysaccharidefrom germi- natedbrownriceby OH method 14 DPPH Figure14 Scavengingefectofpolysaccharidefrom germi- natedbrownricebydpph method 3, PEG6000 PEG6000 15.7% 14.8% 40 min, 81.07%,,DPPH 14,DPPH 81.85%,,, 0.6 mg/ml 68.77%,53.00%,DPPH 81.85% DPPH 2.4.2 (O 2 - [1],,. ) [J].,2014(5):156-160. 15 [2],,,. 15, <0.3mg/mL, [J].,2011,27(6):71-74. 55%, [3]CHO DongHwa,LIM SeungTaik.Germinatedbrownriceand, itsbio-functionalcompounds[j].food Chemistry,2016,196 0.6mg/mL, 68.77%, (8):259-271. [4],,. [J]. 153
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