bpdedna加合物体内外实验研究

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1、四川大学硕士学位论文(themostimportantonewasBPDE)andacid—hydrolysisproductsofDNAadducts(BP-tetr01)weredetectedbyHPLCandconfirmedbyHI’LC-MS．Animalstudies：nlesDratsweretreatedwithsin四ei．P．doseofBaP-DMSO(cosolvent：1％sodiumcarboxymethylcellulose)，150mg／kgratweight．111ebloodoffemoralveinwaswith

2、drawn5hlater,andanticoagulatedusing4％EDTA．n圮bloodDNAWasextractedwithkitandconfirmedtheextractionbyagarosegelelectrophoresis．Afterextraction,theDNAWashydrolyzedin0．1nmol／LHClat90℃waterbathfor4h．Theacid-hydrolysisproducts(BP—tetr01)ofDNAadductswereextractedbyethylacetateandmeasWed

3、byHPLC．FinallyitWasconfirmedbyHPLC-MS．111emeasurememparameters：analyticalcolumn：Phenomenex(USA)，Cn(ODS)，5“m，250x4．6mm；columntemperature：roomtemperature；mobilephaseA：methanol，mobilephaseB：HAc-NH4Ac(pH4．5士o．2)；gradientelutionwithflowrateof1．0ml／min；fluorescencedetector(BPDE：Xd=338

4、um，^．m=425nm；BP—tetrol：k=344nm，A∞=398nm)．11"resultsofHPLCindicatedthattherewereBPDE．DNAadductsandtheformationofBPDEandBP—tetrolwasconfirmedbyHPLC-MS．Invitroresearch，theUVwavelengthofthemaximumabsorptionofDNAafterincubationwithBaPandS9hasbeenredshiftcompared、Ⅳimt}latofDNAstandard

5、whichincubatedinthesamewaybutwithoutBaPandS9fDNAstandard：259．1nm；DNAadducts：305．3nm)，thechangeofspectrumindicatedthatstructureofDNAhadbeenchanged；andthereappearedflewpeaksintheincubationmixturewithBaPandS9intheHPLCchromatogramaswellasintheacid-hydrolysissolution．111enewpeakswere

6、confirmedbyHPLC．MStobeBPDEandBP-tetr01．Inanimalstudies。therewerenewpeaksintheratstreatedwithBaPcomparedwiththecon仃olonesinchromatogram．OneofthenewpeaksWasconfirmedbyHPLC-MStobeBP-tetr01．AccordingtotheresultsofUvspectrum，HPLCandLC·MS，wecouldmakeaconclusionthatundertheexperimental

7、conditions．BaPcouldbetransformedtoBPDE．andtoformBPDE-DNAadductsbothinvitroandvivo．5四川大学硕士学位论文TheHPLCmethodestablishedinthisstudyⅥ髑appliedtomeasurgBPDE．DNAadducts，andfounditwassimple，time—savingandlow-cost．Keywords：UVabsorptionspectrum,Highperformanceliquidchromatography(HPLC)，Hi

8、ghperf．o咖锄celiquidchromatography-massspectromet