稀土离子与药物相互作用的荧光特性研究及应用

华中科技大学硕士学位论文稀土离子与药物相互作用的荧光特性研究及应用姓名：郁丰善申请学位级别：硕士专业：分析化学指导教师：陈芳20080606IStokes(Tb3+)(PUFX)(ATP)(ATP)PUFXTb3+545nm(SDBS)PUFX(Tb3+)(PUFX)PUFX-Tb3+-SDBS6.0×10-8-2.0×10-6mol/L1.0×10-8mol/LPUFX-Tb3+-ATPATPPUFX-Tb3+545nmATPATP4.0×10-7mol/L-2.0×10-5mol/L1.7×10-8mol/LATPIIAbstractRareearthfluorescenceprobecharacterizedwithnarrowspectralwidth,longluminescencelifetime,largestokesshiftandstrongemissionintensity.Itwaswidelyappliedinthemedicineanalysisandbiologicalanalysis.Inthispaper,theinteractionbetweenrareearthionterbium(III)andPUFXwasinvestigatedusingUV–visabsorptionandfluorescencespectra.Ulteriorly,westudiedthetheinteractionsbetweenadenosinedisodiumtriphosphate(ATP)amongthemwerealsoinvestigated.TwonewspectrofluorimetricmethodsweredevelopedfordeterminationofATPandPUFX,respectively.Themainworkissummarizedasfollows:TheemissionspectraofTb3+ionintheprescenceofPUFXandsodiumdodecylbenzenesulfonate(SDBS)werestudied.ItwasfoundthatthecharacteristicpeakofTb3+ionat545nmcouldbegreatlyenhancedbyPUFXandsensitizedSDBS.AndthefluorescenceintensityofPUFX–Tb3+–SDBSsystemwasinproportiontotheconcentrationofPUFX.Accordingly,asensitivefluorimetricmethodfordeterminingprulifloxacinwasproposededbyusingterbium(III)asafluorescenceprobe.Thefluorescenceenhancementmechanismwasalsodiscussedinthetext.Undertheoptimumconditions,thedynamicrangeforthedeterminationofprulifloxacinwas6.0×10-8to2.0×10-6mol/Landthedetectionlimit(3σ)was1.0×10-8mol/L.Theproposedmethodwassimple,accurateandsensitive,andwasappliedsatisfactorilytodetermineprulifloxacininhumanurinewithouttediouspre-separation.TheinteractionsbetweenATPandTb3+-PUFXwereinvestigatedusingfluorescenceandabsorptionspectra.TheresultsindicatedthatthefluorescenceintensityofTb3+-PUFXat545nmcouldbegreatlyenhancedbyATP,andthefluorescenceintensitywasinproportiontotheconcentrationofATP.Basedonwhich,anovelfluorimetricmethodfordeterminingATPwasestablished.Thefluorescenceenhancementmechanismwasalsodiscussedinthetext.Undertheoptimumconditions,thedynamicrangeforthedeterminationofATPwas4.0×10-7to2.0×10-5mol/Landthedetectionlimit(3σ)was1.7×10-8mol/L.Thismethodwassimple,practical,sensitive,anditwasselectiveandsuccessfullyappliedtodeterminationofATPinrealpharmaceuticalsamples.IIIKeywords:spectrofluorimetry;prulifloxacin;rareearthfluorescenceprobe;adenosinedisodiumtriphosphate____________1157La71Lu154f0-145d0-106s1-24f→4fε1015Sm3+Eu3+Tb3+Dy3+1942Wissman[1]β-1.11.1.1[2-7]4f→4f1-11-1Eu3+2[8](1)(2)T1→d(3)1.1.2[9,10](1)4f→4f(2)f→f()()()Eu3+0.2msTb3+2ms(3)Stokes260380nm615nm(Eu)545nm(Tb)643nm(Sm)574nm(Dy)(4)10nm31.2Eu(III)Tb(III)-msEu(III)Tb(III)β-1.2.1(EDTA)HEDDA(N,N-(2-)-N,N-HEDTA(N-2--N,N-DTPA(—N,N,N-)Tb(III)[Tb(EDTA)-ONCH2OCH2NCH2CH2CH2NONCSOOO(1)OCNCOONNCOCCOOHOO(2)1-2EDTA4[Tb(HEDDA)]+[Tb(HEDTA)]0[Tb(DTPA)]2-EDTA(1-2)--(ITEDTA),N-[-]-(ITDTTA,1)(cDTPAA,2)Eu(III)Tb(III)(β-)1.2.22,6-2,2′-2′,2″,2″′-Eu(III)Tb(III)4,7-()-1,10--2,9-(BCPDA,3)[11]4-[2-(4-)]-2,6-[N,N-()]4′-(4-)-2,2′,6′,2″--6′,6″-()-N,N′,N″,N″′-(ITTPBTA,4),4′-(3--4-)-6′,6″-()]-2,2′,6′,2″-(()(TMT,5)πTb(III)Tb(III)Tb(III)Eu(III)BCPDANEu(III)BCPDATMT[12]TMT-Eu(III)TMTEu(III)9(ε287=26970ε319=15278,pH9.0)λex/λem=325nm/613nmTMT-Eu(III)3×10-17mol/LTMT-NH2DNA3′5′TMT-Eu(III)DNA5NNSO2ClClO2SCOOHHOOC(3)NCSNNNNNCOO-COO-COO-COO-(4)OCH3NNH2NNNNCOO-COO-COO-COO-(5)1-361.2.3β-β-R1-CO-CH2-CO-R2[13]β-(HNF3HNF5HNF7HPF3HPF5)R12-2-R2(C4H9C3H7C2H5CF3)Yuan[14]TOPO()1-11-1-β-λmax/nmε×104(cm-1mol-1L)λex/nmλem/nmФfτ/µs[Eu(NF3)3(TOPO)2]3395.823396150.209920[Eu(NF5)3(TOPO)2]3396.213396150.260519[Eu(NF7)3(TOPO)2]3396.393396150.335530[Eu(NF9)3(TOPO)2]3396.583396150.325530[Eu(PF3)3(TOPO)2]3446.583446150.127444[Eu(PF5)3(TOPO)1.5]3469.003466150.178360[Eu(PF7)3(TOPO)1.5]3469.073466150.266378[Eu(PF9)3(TOPO)1.5]3469.343466150.272387R2R1Фf2-2-;R1R2ФfC4H9C3H7C2H5CF3β--β-Yuan[15,16]4,4-(1,1,1,2,2,3,3--4,6-)-O-(BHHCT,6)4.5×10-13mol/L7C3F7OOOOC3F7ClSO2(6)1-3BHHCT1.2.4[2.2.1](7)(8)(9)1-4Sabbatini[17](1-4(7))[Eu(III)-2.2.1]3+-2F-1.01.0ms[Tb(III)-2.2.1]3+-2F-3.1msSabbatini[17](8)Tb(III)Eu(III)Eu(III)0.170.40msPart[18](8)Eu(III)-Eu(III)Eu(III)(9)Eu(III)Tb(III)pH6.0Tris-BCPDAStokesλem(Eu(III))=619594nmλem(Tb(III))=548nm[19]Tb(III)Eu(III)8OOONOON(7)NNNNNNCH2COOHCH2COOH(9)NNNNNNO-ONNCOOCH3COOCH3(8)1-491.31.3.1Stokes10-1000µs1-20ns1-5Tb(III)-NFLX(1)0.50µg/mlNFLX(pH8.0)(2)0.50µg/mlNFLX+4.92×10-5µg/mlTb(III)(pH8.0)(3)0.50µg/mlNFLX+4.92×10-5µg/mlTb(III)(pH8.0)+4.92×10-5µg/mlSDSHirschy[20]Eu(III)10[Eu(III)Tb(III)]pH7.010Tb(III)(NFLX)(CPFX)(ENX)(GENX)(1-5)SDSTb(III)-NFLXSDSTb(III)-ENXTb(III)-GENXNFLX[21]1-21-2(DL)Eu3+Eu3+-20Eu3+Eu3+-Triton-X100(pH7.5)DL3.0×10-8mol/L22Eu3+Eu3+--TC,0.1-10µg/ml;OTC,0.15-5.0µg/ml23Eu3+Triton-X100,TOPO-Eu3+HPLC24Eu3+-Eu3+(pH7.5)DL1.1×10-5mol/L25Tb3+Tb3+26Tb3+Tb3+DL1.0×10-8mol/L27Tb3+Tb3+--EDTA28L-Tb3+Tb3+-L-Dopa-SDS1.1×10-8-5×10-6mol/L,DL5.0×10-9mol/L29Tb3+HPLCTb(III)0.01-1.0ng/g30(NFLX)Tb3+Tb3+-NFLX-SDSDL0.017µg/ml211.3.2Eu(III)Tb(III)304nm11A304nmB()()BBreen[31]Tb(III)5D4-7FJEu(III)5D0-7FJ4f→4fEu(III)Tb(III)Eu(III)Tb(III)Austin[32]Tb(III)()Eu(III)Tb(III)Tb(III)()Tb(III)Brittain[33]Tb(