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3(,100085)(H2O2)(1O2)(O-2)(OH),,,,2001211208;2002205205(No.20125514)1,,,1770JosephPriestley,,,,,,,,1969McCordFridovich1,O2(O-2),O-2,(superoxidedismultase,SOD)O-2,SODO-2(H2O2)(OH)(,1O2)10,,,,28,,,,,,,:,(1O2);3,(O-2)(OH)(HO2);2,(H2O2)(ROOH)(NO),1O22sOH200sO-25s,H2O2,,936374748593,2,,,,91621,1722232530200212(FENXIHUAXUE)ChineseJournalofAnalyticalChemistry121507151426293036,,,,2.1,6063,H2O2,6468,H2O26972,(luminol)(lucigenin)cypridinaluciferinanalog(CLA),(pH10),,,,Fe2+Co2+Ni2+Cu2+,(APD),APD,73,741928Albrecht75,70,,,,BaO2MgO2,76,ABEI77,782,,71,,,APD,,H2O2O-2OH1O2,,,,GleuPetsch791935,H2O2,O-2N2(N2methylacridan),,76,80,81,,H2O2O-2,,22methyl262phenyl23,72dihydroimidazo21,22apyrazin232one,CLA,1970Goto82CLAH2O2OH,O-21O2,380nm838486Goto87CLA,MCLAFCLANakanoFujimori8890,CLA,MCLAFCLA(532nm),CLA10,,O-21O2,OH2.2,(cytochromeC)1,20(nitrobluetetrazolium,NBT)92CO-2,550nm93,O-2,SOD,O-2SODH2O2O2,SOD94,,HADPA,O-2,SOD,O-2,O-2,1,NBTO-2,(diformazan)92,95,8051301Fig.1Colorchangeofthereactionofnitrobluetetrazoliumwithsuperoxideanionicradicalion560nm,10,000,KIO42H2O270,NBTO-2,,,,O-2SutherlandLearmonth96NBTO-2,H2O2TiD97,(HRP)20,21982.3,,,,,(DCFH),H2O2HO-2DCF,H2O2232,32(DAN)NO-2,12(H)2,2,32(DAN)NO24,Kojima25NO222(DAF222DA),5nmolPLNO,2.4(ESR)ESR,,,,ESRUtsumi27,,O-2,OH(microsome),,,ESR,TMP(2,2,6,62tetramethyl242piperidone),1O2,,1O226,,1O2,,ESR,29,992.5,2NO30,NO10-20mol,H2O2,LvovichScheeline33O-2H2O2,,(polypyrrole)PHRP,H2O2;,PSOD,O-2,pH5.19.0,,,,,905112:,,3,,,,(HPLC)(GC),(spintrap),,,,5,52212(5,52dimethylpyrroline212oxide,DMPO),,ESR3.1Smith100,101,1O2,52102,7272,HPLC,391O21O2,,,14103,1O2104OH,OH2,322,5237,HPLC,,OH41Kamiya40OH1O282(82OH2Gua),HPLC82,,,3.22,DMPOPBM(2pheny2N2tert2butynitrone),,ESR43DMPO23Fig.2BasicprincipleofspintrappingmethodandthreeimportantspintrapsDMPO:5,52212(5,52dimethyl212pyrroliue212oxide;TMPO:3,3,5,52212(3,3,5,52tetramethyl212pyrroliue212oxide);DBNBS:3,52242(3,52dibromo242nitrosobenzenesulfonate),,DMPOPBNO-2,OH,ESR,O-2OHESR(hfs:hyperfinesplitting,)DMPOPBN,ESRg,O-2OH,,O-2DMPO2OOH,OHDMPO2OHESR,O-2,OH,Kohno45(DMSO),Rouband46DMPO015130,DEPMO52(dithoxyphosphoryl)252methyl2pyrroline2oxide,O-2(15min),12ESR,,,ESR,4,1O2ESR4.11O2,1O2,16+g1g,,1g4.2s105,16+g3Fig.3Oxygensingletmoleculestateandsimultaneoustransitionmolecular2pairstates,1O210-3molPL106,10-10s,,,,1g1O2,1g,1+g1g1O2,,3,634nm762nm1O250,1268nm1O251,1O253,54,56,574.2O-2,OH4Fig.4Electronspinresonance(ESR)spectraofsuperoxideradicals1177K(thesuperxideradicaliongeneratedfromthereactionofxanthinewithxanthineoxidaseat77K);2177K(thesuperoxideradicaliongeneratedfromthereactionofH2O2withNaIO4at77K),ESR,,ESR,,,,ESR(77K),4O-2ESR58,O-2,ESR,ggg,ESR,4,ESR,,ESR,115112:5,,,,invivo,,,DNA,invivo899References1McCordJM,FridovichIJ.Biol.Chem.,1969,244:60492SiesH.OxidativeStress.London:AcademicPress,19853YaguiK,NakanoM.ActiveOxygenSpecies.Tokyo:YihayakuPublishingHouse,19874AfanasevIB.SuperoxideIon2ChemistryandBiologicalImmplications.Florida:CRCPress,19895ChemistryofOxygenSpecies(KikanKagakuSosetu).Tokyo:TheChemicalSocietyofJapan,19906SawyerDT.OxygenChemistry.NewYork:OxfordUniversityPress,19917NakanoM,AsadaK,OyanakiS.PND(Jpn.J.Protein,NucleicAcidandEnzyme),1988,33(Suppl.):265331888NakanoM,YoshikawaT.ActiveOxygenSpeciesandLuminescence.Tokyo:NipponYikagakuKanPress,19909SugiokaK,NakanoM,KurashigeS,AkuzawaY,GotoT.FEBSLett.,1986,197:2710TeranishiK,ShimomuraO.Anal.Biochem.,1997,249:3711MurphyME,SiesH.Anal.Proc.,1990,27:21721912TawaR,SakuraiH.Anal.Lett.,1997,30:2811282513NosakaY,YamashitaY,FukuyamaH.J.Phys.Chem.B,1997,101:5822582714LundbergJON,LundbergJM,AlvingK,WeitzbergE.NatureMed.,1997,3:2015MikuskaP,VeceraZ.Anal.Chim.Acta,1998,374:29730216PosnerGH,LeverJR,MiuraK,LisekC,SeligerHH,ThompsonA.Biochem.Biophys.Res.Commun.,1984,123(2):86987317HiguchiK,MotomizuS.Anal.Sci.,1999,15:12913418ClappPA,EvansDF,SheriffTSS.Anal.Chim.Acta,1989,218:33133419SarkerAK,UkedaH,KawanaD,SawamuraM.Anal.Sci.,1999,15:1141114420BoverisA,OshinoN,ChanceB.Biochem.J.,1972,128:61763021ClappPA,EvansDF.Anal.Chim.Acta,1991,243:21722022LiuZ,CaiR,MaoL,HuangH,MaW.Analyst,1999,124:17317623BassDA,ParceJW,DeChateletJR,SzejdaP,SeedsMC,ThomasMJ.Immunol.,1983,130:191024MiskoTP,SchillingRJ,SalveminiD,MooreWM,CurrieMG.Anal.Biochem.,1993,214:111625KojimaH,NakatsuboN,KikuchiK,KawaharaS,KirinoY,NagoshiH,HirataY,NaganoT.Anal.Chem.,1998,70:2446245326LionY,DelemlleM,VorstA.Nature,1976,263:44244327UtsumiH,ShimakuraA,KashiwagiM,HamadaA.J.Biochem.,1989,105:23924428KawaneM,IidaT,YoshikiY,OkuboK,TsunakawaM.Luminescence,1999,14:32132529OzawaT.Bunseki(Jpn.J.Anal.),2000,(2):687430MalinskiT,TahaZ.Nature,1992,358:67667831AoyamaY,SakuriS.NipponKagakuKaishi(Jpn.J.Chem.Soc.Japan),1997,(7):47948332ZamboninCG,LositoI.Anal.Chem.,1997,69:4113411933LvovichV,ScheelineA.Anal.Chem.,1997,69:45446234XiaoY,JuHX,ChenHY.Anal.Chim.Acta,1999,391:29930635LisdatF,GeB,Ehrentreich2ForsterE,ReszkaR,SchellerFW.Anal.Chem.,1999,71:1359136521513036XiaoY,JuHX,ChenHY.Anal.Chim.Acta,1999,391:738237FloydRA,WatosonJJ,WongPK.J.Biochem.Biophys.Methods,1984,10:22138HusainSR,CillardJ,CillardP.Phyto