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1、抗氧化方法一、DeterminationofSuperoxideAnionScavengingAbility(超氧陰離子清除能力)1、determinedbyaCLmethodinthepyrogallol-luminolsystemonaBPCLUltra-weakluminescenceanalyzer。(焦性沒食子酸-發(fā)光胺,熒光檢測)2、步驟:10mLsample(不同濃度)+50mL焦棓酸(6.25*10-4mol/L)+0.94mLofamixturecontainingluminol(0.05mol/L)andcarbonatebuffer(pH10.2)(發(fā)
2、光胺用pH10.2碳酸鹽緩沖液配成0.05mol/L)3、Hi-V,800;Kv,-1;thespectralrangeofCL(波長范圍)180-800nm;溫度:30°C.總時間:300S,每2S讀數(shù)一次。4、對照:不加樣品(樣品用水代替)??瞻撞患咏箺斔幔ㄓ脕碚{(diào)零)。二、DeterminationofScavengingAbilityonHydroxideRadicals(羥基自由基清除能力)1、CuSO4-Phen-Vc-H2O2CLsystem(1mL體系)2、50mLofsamplesolution(樣品液)+50mLofa1.0mmol/LCuSO4solut
3、ion(CuSO4溶液)+50mLofa1mmol/L1,10-phenanthrolinesolution(鄰二氮菲溶液)+700mLofa0.05mol/Lboratebuffer(pH9.0)(硼酸緩沖液)+100mLofa1mmol/Lascorbatesolution+50mLofa0.15%H2O2solution3、總時間400S,間隔3S。Hi-V,800;Kv,-1;thespectralrangeofCL(波長范圍)180-800nm;溫度:30°C.4、陽性對照:不加樣品(樣品用水代替)。空白不加H2O2(用來調(diào)零)。三、Determinationof
4、ScavengingEffectonHydrogenPeroxide(過氧化氫清除能力)1、luminol-H2O2system2、Theluminescentreactionwasinitiatedbymanuallyadding1mLofasolutioncontaining0.15mol/Lhydrogenperoxideand0.1mol/Lluminolperliterofcarbonatebuffer(0.05mol/L,pH9.4).Lightemissionvstimewasrecordedfor3minat2sintervals.步驟:0.15mol/L的
5、過氧化氫+0.1mol/L發(fā)光胺(用pH9.4的碳酸緩沖液配),總體系1ml.3、BPCLUltra-weakluminescenceanalyzer,Hi-V,800;Kv,-1;thespectralrangeofCL(波長范圍)180-800nm;溫度:30°C.4、Thecontrolwasperformedinthesamemannerinthemixturewithoutthesamplesolution,andthebackgroundwasdetectedwithoutH2O2addition。三、DeterminationofPreventingDNADa
6、mageEffect(DNA損傷)1、CuSO4-Phen-Vc-H2O2-DNACLsystem.2、步驟:Copperand1,10-phenanthrolinewerepremixedin0.1mol/LNaOAc/HOAc(pH5.5)buffer。3mg/mLDNAwasincubatedwithaphen-Cusolution.步驟:800mLofphen-Cu/DNAsolution+100mLof4.2*10-3mol/Lascorbate+200mLof6%H2O2+wereaddedwithoutintervaltoa100mLsamplesolutio
7、ntogiveafinalvolumeof1.2mL.(總體系1.2mL)3、ThekineticcurveofCLproducedinthephen/Cu/H2O2/ascorbatesystemwasimmediatelyrecorded.(Thecontrolwasperformedinthesamemannerinthemixturewithoutthesamplesolution,andthebackgroundwasdetectedwithoutH2O2addition.)4、1.0mmol/LCuSO