title: Ozonation of DNA forms adducts: a 32P-DNA labeling and thin-layer chromatography technique to measure DNA environmental biomarkers.
abstract: Little direct documented evidence of ozone's genotoxicity exists. Deoxyribonucleic acid (DNA) adducts are produced by environmental toxic agents, including ozone. We have described a modified thin-layer chromatography (TLC) technique that can assess adduct formation as a biomarker of ozone injury. This requires 32P-labeling DNA, digestion of deoxynucleotides (dNMPs), and separation in two-dimensional PEI-cellulose TLC. We have applied this technique to control DNAs, to control DNA in solution exposed to acute ambient ozone, and to control DNA exposed to acute bubbled-through ozone (2 ppm for 24 h). We detected stable DNA adducts, including hydroxymethyluracil (HMU), thymine glycol (TG), 8-hydroxyguanine (8-OHG), and demonstrated, as yet, unidentified adducts that may serve as a "fingerprint" pattern of DNA adduction. This technique quantifies low-molecular-mass DNA adducts, both in vivo and in vitro, with potential applications to environmental toxicology.
authors.affiliation: Department of Pathology, Albert Einstein College of Medicine, New York, New York.
authors.forename: A
authors.initials: A
authors.lastname: Cajigas
authors.forename: M
authors.initials: M
authors.lastname: Gayer
authors.forename: C
authors.initials: C
authors.lastname: Beam
authors.forename: J J
authors.initials: JJ
authors.lastname: Steinberg
mesh_terms: D000818:Animals; D001345:Autoradiography; D015415:Biomarkers; D002855:Chromatography, Thin Layer / Q000379:methods; D004247:DNA / Q000032:analysis*; D004249:DNA Damage; D006367:HeLa Cells; D006801:Humans; D010126:Ozone / Q000633:toxicity*; D010761:Phosphorus Radioisotopes
chemical_list: D015415:Biomarkers; D010761:Phosphorus Radioisotopes; D010126:Ozone; D004247:DNA