Archives of Toxicology

, Volume 93, Issue 3, pp 763–773 | Cite as

Evaluation of inhaled low-dose formaldehyde-induced DNA adducts and DNA–protein cross-links by liquid chromatography–tandem mass spectrometry

  • Jiapeng Leng
  • Chih-Wei Liu
  • Hadley J. Hartwell
  • Rui Yu
  • Yongquan Lai
  • Wanda M. Bodnar
  • Kun LuEmail author
  • James A. SwenbergEmail author
Genotoxicity and Carcinogenicity


As a widespread industrial chemical, formaldehyde carcinogenicity has been highly controversial. Meanwhile, formaldehyde is an essential metabolite in all living cells. Previously, we have demonstrated exogenous formaldehyde causes DNA adducts in a nonlinear manner between 0.7 and 15.2 ppm using [13CD2]-formaldehyde for exposure coupled with the use of sensitive mass spectrometry. However, the responses from exposure to low doses of formaldehyde are still unknown. In this study, rats were exposed to 1, 30, and 300 ppb [13CD2]-formaldehyde for 28 days (6 h/day) by nose-only inhalation, followed by measuring DNA mono-adduct (N2-HOMe-dG) and DNA–protein crosslinks (dG-Me-Cys) as formaldehyde specific biomarkers. Both exogenous and endogenous DNA mono-adducts and dG-Me-Cys were examined with ultrasensitive nano-liquid chromatography–tandem mass spectrometry. Our data clearly show that endogenous adducts are present in all tissues analyzed, but exogenous adducts were not detectable in any tissue samples, including the most susceptible nasal epithelium. Moreover, formaldehyde exposure at 1, 30 and 300 ppb did not alter the levels of endogenous formaldehyde-induced DNA adducts or DNA–protein crosslinks. The novel findings from this study provide new data for risk assessment of exposure to low doses of formaldehyde.


Formaldehyde DNA damage DNA adducts DNA–protein crosslinks Mass spectrometry 







DNA–protein crosslinks


Higher-energy collisional dissociation


Tandem mass spectrometry


Parallel reaction monitoring


Selected reaction monitoring



We thank Dr. Leonard Collins for his assistance with HPLC purification and nano-UPLC–MS–MS. This project received funding from the American Chemistry Council and Formacare. The instrumentation was partially supported by funding from the NIEHS (R01ES024950 and P30ES010126). None of the funding agencies is involved in writing or have access to the manuscript before its submission.

Compliance with ethical standards

Conflict of interest

The authors declare they have no actual or potential competing financial interests.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Jiapeng Leng
    • 1
    • 2
  • Chih-Wei Liu
    • 1
  • Hadley J. Hartwell
    • 1
  • Rui Yu
    • 1
  • Yongquan Lai
    • 3
  • Wanda M. Bodnar
    • 1
  • Kun Lu
    • 1
    Email author
  • James A. Swenberg
    • 1
    Email author
  1. 1.Department of Environmental Sciences and EngineeringUniversity of North Carolina at Chapel HillChapel HillUSA
  2. 2.School of Chemical Engineering and Technology, North University of ChinaTaiyuanChina
  3. 3.Lovelace Respiratory Research InstituteAlbuquerqueUSA

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