NanoLC/ESI+ HRMS3 quantitation of DNA adducts induced by 1,3-butadiene

Dewakar Sangaraju, Peter W. Villalta, Susith Wickramaratne, James Swenberg, Natalia Tretyakova

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

(Chemical Presented) Human exposure to 1,3-butadiene (BD) present in automobile exhaust, cigarette smoke, and forest fires is of great concern because of its potent carcinogenicity. The adverse health effects of BD are mediated by its epoxide metabolites such as 3,4-epoxy-1-butene (EB), which covalently modify genomic DNA to form promutagenic nucleobase adducts. Because of their direct role in cancer, BD-DNA adducts can be used as mechanism-based biomarkers of BD exposure. In the present work, a mass spectrometry-based methodology was developed for accurate, sensitive, and precise quantification of EB-induced N-7-(1-hydroxy-3-buten-2-yl) guanine (EB-GII) DNA adducts in vivo. In our approach, EB-GII adducts are selectively released from DNA backbone by neutral thermal hydrolysis, followed by ultrafiltration, offline HPLC purification, and isotope dilution nanoLC/ESI+-HRMS3 analysis on an Orbitrap Velos mass spectrometer. Following method validation, EB-GII lesions were quantified in human fibrosarcoma (HT1080) cells treated with micromolar concentrations of EB and in liver tissues of rats exposed to sub-ppm concentrations of BD (0.5-1.5 ppm). EB-GII concentrations increased linearly from 1.15±0.23 to 10.11±0.45 adducts per 106 nucleotides in HT1080 cells treated with 0.5-10 μM DEB. EB-GII concentrations in DNA of laboratory rats exposed to 0.5, 1.0, and 1.5 ppm BD were 0.17±0.05, 0.33±0.08, and 0.50±0.04 adducts per 10 6 nucleotides, respectively. We also used the new method to determine the in vivo half-life of EB-GII adducts in rat liver DNA (2.20±0.12 d) and to detect EB-GII in human blood DNA. To our knowledge, this is the first application of nanoLC/ESI+-HRMS3 Orbitrap methodology to quantitative analysis of DNA adducts in vivo.

Original languageEnglish (US)
Pages (from-to)1124-1135
Number of pages12
JournalJournal of the American Society for Mass Spectrometry
Volume25
Issue number7
DOIs
StatePublished - Jul 2014

Keywords

  • 1,3-butadiene (BD)
  • Accurate mass
  • DNA adducts
  • Orbitrap mass spectrometry
  • Quantitation

Fingerprint Dive into the research topics of 'NanoLC/ESI<sup>+</sup> HRMS<sup>3</sup> quantitation of DNA adducts induced by 1,3-butadiene'. Together they form a unique fingerprint.

Cite this