32P-Postlabeling high-performance liquid chromatographic improvements to characterize DNA adduct stereoisomers from benzo[a]pyrene and benzo[c]phenanthrene, and to separate DNA adducts from 7,12-dimethylbenz[a]anthracene.
Single compounds can generate complex DNA adduct patterns by reactions through different pathways, with different target nucleotides and through different configurations of the products. DNA adduct analysis by 32P-HPLC was improved by adding an isocratic plateau in an otherwise linear gradient, thereby enhancing resolution of predictable retention time intervals. This enhanced 32P-HPLC technique was used to analyze and at least partly resolve 14 out of 16 available benzo[c]phenanthrene deoxyadenosine and deoxyguanosine adduct standards, 8 out of 8 available benzo[a]pyrene deoxyadenosine and deoxyguanosine adduct standards, and 51 peaks from 7,12-dimethylbenz[a]anthracene-calf thymus DNA reaction products. The same type of gradient modifications could be used to enhance resolution in analyses of other complex DNA adduct mixtures, e.g., in vivo in humans.
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Karolinska Institutet, Department of Bioscience, NOVUM Research Park, Huddinge, Sweden.
Möller
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D015127:9,10-Dimethyl-1,2-benzanthracene / Q000031:analogs & derivatives* / Q000032:analysis; D000818:Animals; D001564:Benzo(a)pyrene / Q000032:analysis*; D002273:Carcinogens; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D018736:DNA Adducts / Q000032:analysis*; D003839:Deoxyadenosines; D003849:Deoxyguanosine; D009153:Mutagens; D010616:Phenanthrenes; D010761:Phosphorus Radioisotopes; D015203:Reproducibility of Results; D013237:Stereoisomerism
C059444:7,12-dimethylbenz(a)anthracene-DNA adduct; D002273:Carcinogens; D018736:DNA Adducts; D003839:Deoxyadenosines; D009153:Mutagens; D010616:Phenanthrenes; D010761:Phosphorus Radioisotopes; C056612:benzo(a)pyrene-DNA adduct; D001564:Benzo(a)pyrene; D015127:9,10-Dimethyl-1,2-benzanthracene; D003849:Deoxyguanosine; C002615:benzo(c)phenanthrene