Analytical and Bioanalytical Chemistry

, Volume 393, Issue 5, pp 1525–1530 | Cite as

Ultrasensitive method for the determination of 4-hydroxy-1-(3-pyridyl)-1-butanone-releasing DNA adducts by gas chromatography–high resolution mass spectrometry in mucosal biopsies of the lower esophagus

  • Christopher W. Heppel
  • Anne-Kathrin Heling
  • Elmar RichterEmail author
Original Paper


4-Hydroxy-1-(3-pyridyl)-1-butanone (HPB)-releasing DNA adducts are formed by metabolic activation of the tobacco-specific nitrosamines 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N′-nitrosonornicotine (NNN). NNK and NNN are considered carcinogenic to humans by the International Agency for Research on Cancer. Existing analytical methods for determination of HPB-releasing DNA adducts require 0.3–2.0 g of human target tissues such as lung and esophagus. For adduct determination in milligram amounts of biopsy samples, an ultrasensitive and specific method is presented using capillary gas chromatography coupled to a high-resolution mass spectrometer operated in the negative chemical ionization mode (GC-NCI-HRMS). The method has a limit of detection of 4.6 fmol HPB, a limit of quantification of 14.9 fmol HBP and a recovery of 45 ± 15%. Intra- and inter-day imprecision for N = 6 samples were calculated with coefficients of variation of <3.1%. Method applicability was evaluated with biopsies of esophageal mucosa (N = 14) yielding 5.6 ± 1.9 mg tissue and a mean adduct level of 6.13 ± 9.35 pmol HPB/mg DNA.


Esophagus High-resolution GC-NCI-HRMS Tobacco-specific nitrosamines DNA adducts 



Research described in this article was supported by Philip Morris USA Inc. and Philip Morris International.


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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Christopher W. Heppel
    • 1
  • Anne-Kathrin Heling
    • 1
  • Elmar Richter
    • 1
    Email author
  1. 1.Walther Straub Institute, Department of ToxicologyLudwig-Maximilians UniversityMunichGermany

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