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24,5Vol124,No15,pp596259920045SpectroscopyandSpectralAnalysisMay,2004,,,510631,,(:),,,Eu3+,:336337nm,Eu3+;;;Eu3+:O657132:A:100020593(2004)0520596204:2003203226,:2003206228:(2002C60113);(2002XGP06);(No1015012,No.031518);(No102113):,1965,,(TR2FIA),1983Petterson[1]Eskola[2],TRFIA10-19molwell-1,(EIA)10-9molwell-1,(RIA)10-15molwell-1(LIA)10-15molL-1,,,,22(),,,11s22s22p63s23p63d104s24p64d104fn5s25p6(n=014),,4fn(n=014),4fn-15d[3][4]111fffn,J:(1)ff,,,dd,(2)4f5s2,5p6,(3),()112fd4fn4fn-15d,4f5d;;5d,ff,113,,(Sm3+,Eu3+,Te3+,Yb3+Ce4+)2211,,,,,,:ff;5d4f;,,4fn-4fn-15d,4fn,,212[4],(1):(1),S0S1,S0(),T1T2;(2),(),();(3)(),,,,Fig11Energylevelschemeoftheenergy2lossprocedureoftheexcitedstate,,,,,,,,,,,,,,,,,Eu3+Eu2+,Eu3+,[5](2),Eu3+Cl-,Br-,I-301,23415,292,189nmFig12Charge2transfertransitionspectrumofEu3+halide(1)Eucl-36;(2)EuBr-363Eu3+Eu3+5D07FJ,(4f),5D07F1,7F2,7F4590596nm,610620nm,687703nm2Eu3+,,Eu3+5DJ,Eu3+,Eu3+5D07F2(610620nm),Eu3+5D07F24f5d5D07F24TRFIAEu3+TRFIA,,Eu3+Fig13AbsorptionspectraofthefluorescentEu3+chelate(L1C,Labellingchelate;E1L,Enhancementliquid)7955Eu3+300350nm(3),,5D07F2,Eu3+5D0,300350nm,4Fig14FluorescencespectrumofthefluorescentEu3+chelate4Eu3+,336337nm(610620nm),(5):336337nm,,Fig15ThehighesteffectoffluorescenceenhancementbythefluorescentEu3+chelate(300nm)Eu3+(6),Eu3+5DJ,5D07FJ,220280nm,4f,,66,4f(4f5d),Eu3+Fig16Fluorescencespectraofthelabelingchelate5TRFIA,,,101000s,,,TRFIA,Eu3+(),,Eu3+,:2EDTA,2EDTA,12(P2)2EDTA,,,,TRFIA,,pH,232Eu3+(336337nm),,Eu3+5D07FJ(610620nm)1987,Lehn[6]Eu,,,LehnGerardMathisXL665(allophycocyanin),XL665,,,89524Fig17Principlesofhomogeneoustime2resolvedfluorescence77337nm,,XL665,665nm,,665nm,7,XL665(915nm[6],665nm,,7,XL665,620nm,665nm6TRFIA,20,,,[7,8][1]PettersonKetal.Clin.Chem.,1983,29:60.[2]EskolaJuetal.Clin.Chem.,1983,29:1777.[3]MichelsenB.AnalysisandApplicationofRareEarthMaterials.Oslo:Universitetsforlaget,1973.[4]ZHANGRuo2hua().LanthanideChemistry().TianjinPubhishingCompanyofScienceandTechnology(),1987.[5]RyanJL.J.Phys.Chem.,1966,70:2845.[6]DietrichB,SauvageJ2P.TheNobelAwardGoestoCrownEthers,CryptandsandOtherHost3GuestMolecularSystems.NewJ.Chem.,1987,12:8.[7]TIANZhen,GUOZhou2yietal(,).ActaLaserBiologySinica(),2002,11(4):290.[8]TIANZhen,GUOZhou2yi(,).J.OptoelectronicsLaser(),2002,13(11):1998.SpectroscopicEvaluationofTime2ResolvedFluoroimmunoassayGUOZhou2yi,TIANZhen,JIAYa2liInstituteofLaserLifeScience,SouthChinaNormalUniversity,Guangzhou510631,ChinaAbstractThelanthanidetrivalenceionanditschelatesareusedformarkingsubstanceinTime2ResolvedFluorescenceImmunoassay(TRFIA),markingprotein,hormone,antibody,nucleicacidprobeorbiologicalivecell,tomeasuretheconcentrationoftheanalysissubstanceinsidethereactionsystemwithtime2resolvedfluorometryafterthereactionsystemoccurred,andattainthequantitativeanalysisspurpose.TRFIAhasthereforebecomeakindofnewandmoresensitivemeasurementmethodafterradioisotopemarking,enzymaticmarking,chemiluminescence,electrochemiluminescence,primarilydependingonthespecialphysicsandchemistrycharacter2isticsoflanthanidetrivalenceionanditschelates.Inthispaper,theresultofspectroscopicevaluationofeuropiumtrivalenceionanditschelate,andtheprincipleoftime2resolvedtechnologyandfluorescence2enhancedtechnologyarereported.Atthesametime,theexper2imentshowsthattheexcitationwavelengthchosenbetween336and337nmbenefitstheexcitationandtheenergytransferofchelatediketoneofeuropiumtrivalenceion.KeywordsImmunoassay;Fluorescence2enhancedtechnology;Time2resolvedspectroscopictechnology;Europiumtrivalenceionchelate(ReceivedMarch26,2003;acceptedJune28,2003)9955