Ozonation of dna forms adducts: A 32p-dna labeling and thin-layer chromatography technique to measure dna environmental biomarkers

Antonio Cajigas, Mitchell Gayer, Carl Beam, J. J. Steinberg

Research output: Contribution to journalArticle

12 Scopus citations

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 (TC), 8-hydroxyguanine (8–OHG), and demonstrated, as yet, unidentified adducts that may serve as a “fingerprint” pattern of DNA adduction. This technique quantifies low-molecularmass DNA adducts, both in vivo and in vitro, with potential applications to environmental toxicology.

Original languageEnglish (US)
Pages (from-to)25-36
Number of pages12
JournalArchives of Environmental Health
Volume49
Issue number1
DOIs
Publication statusPublished - Jan 1 1994

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ASJC Scopus subject areas

  • Environmental Chemistry
  • Environmental Science(all)
  • Public Health, Environmental and Occupational Health
  • Health, Toxicology and Mutagenesis

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