Chromatographic conditions for separation of 32P-labeled phosphates of major polynuclear aromatic hydrocarbon--deoxyribonucleoside adducts.
The 3'-monophosphates of the major deoxyribonucleoside adducts of five representative polynuclear aromatic hydrocarbons (PAH)-benzo[a]pyrene, benz[a]anthracene, chrysene, 5-methylchrysene and 6-methylchrysene--were prepared by reaction with DNA of the appropriate bay-region diol epoxides followed by enzymatic hydrolysis and HPLC purification. These standards were converted to 3',[32P]-5'-bisphosphates as in the Randerath 32P-postlabeling assay. TLC and HPLC systems for their separation are described. HPLC conditions for separation of the corresponding 3'- and 5'-monophosphates are also reported. These data will be useful for determining whether unknown DNA adducts detected by the 32P-postlabeling assay may have resulted from reactions of DNA with PAH diol epoxides.
Amin
S
S
Division of Chemical Carcinogenesis, American Health Foundation, Valhalla, NY 10595.
Misra
B
B
Desai
D
D
Huie
K
K
Hecht
S S
SS
D004247:DNA; D003854:Deoxyribonucleotides; D004852:Epoxy Compounds; D015394:Molecular Structure; D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds; D011865:Radioisotope Dilution Technique; D013329:Structure-Activity Relationship
D003854:Deoxyribonucleotides; D004852:Epoxy Compounds; D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds; D004247:DNA