Detection of 1,N2-propanodeoxyguanosine adducts of trans-4-hydroxy-2-nonenal after gavage of trans-4-hydroxy-2-nonenal or induction of lipid peroxidation with carbon tetrachloride in F344 rats

Chem Biol Interact. 2001 Sep 28;137(3):269-83. doi: 10.1016/s0009-2797(01)00259-9.


The 1,N2-propanodeoxyguanosine adducts of trans-4-hydroxy-2-nonenal (HNE-dGp-adducts) were quantitated in tissues of rats treated with trans-4-hydroxy-2-nonenal (HNE) or carbon tetrachloride, respectively, using a 32P-postlabeling method. The method development was based on chemically synthesized HNE-1,N2-propanodeoxyguanosine adduct standard, which was characterized by NMR and mass spectra. The adducts were enriched by Nuclease P1. They were subsequently reacted with gamma-32P-ATP to give the respective 3'-5'-bisphosphates, which were two-directionally separated on PEI-cellulose-TLC and quantitated by autoradiography. The labeling efficiency for the adduct standard was 27%, and the recovery of spiked amounts of adduct standard in the enzymatical procedure was about 80%. Internal standard was used to eliminate methodological variations. The determination of the limit of quantitation in DNA from rat tissues by spiking of HNE-dGp-adduct standard revealed a sensitivity of about 20 HNE-dGp-adducts/10(9) normal nucleotides. Background levels of HNE-dGp-adducts in tissues of rats including liver, kidney, lung, colon and forestomach were found in the range of 18-158 adducts/10(9) nucleotides with relatively high adduct levels in the liver and low adduct levels in kidney, lung and colon. These background levels were statistically significantly increased by the factor of 2 in liver, lung, colon and forestomach after induction of lipid peroxidation by carbon tetrachloride. The finding that background HNE-dGp-adduct levels may be in context with different metabolic activities of the tissues and the increase of HNE-dGp-adduct levels after application of carbon tetrachloride indicate that HNE-dGp-adducts are an endogenous lesion and that they are probably formed from radical initiated lipid peroxidation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Oral
  • Aldehydes / administration & dosage
  • Aldehydes / metabolism
  • Aldehydes / pharmacology*
  • Animals
  • Autoradiography
  • Carbon Tetrachloride / administration & dosage
  • Carbon Tetrachloride / toxicity*
  • Chromatography, Thin Layer
  • DNA / drug effects*
  • DNA / metabolism
  • DNA Adducts / analysis*
  • DNA Adducts / metabolism
  • Deoxyguanosine / analogs & derivatives
  • Deoxyguanosine / metabolism*
  • Female
  • Injections, Intraperitoneal
  • Lipid Peroxidation / drug effects*
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Rats
  • Rats, Inbred F344
  • Sensitivity and Specificity


  • Aldehydes
  • DNA Adducts
  • 1,N(2)-propanodeoxyguanosine
  • DNA
  • Carbon Tetrachloride
  • Deoxyguanosine
  • 4-hydroxy-2-nonenal