Journal Article

Ethylation and methylation of hemoglobin in smokers and non-smokers

Steven G. Carmella, Menglan Chen, Peter W. Villalta, James G. Gurney, Dorothy K. Hatsukami and Stephen S. Hecht

in Carcinogenesis

Volume 23, issue 11, pages 1903-1910
Published in print November 2002 | ISSN: 0143-3334
Published online November 2002 | e-ISSN: 1460-2180 | DOI:
Ethylation and methylation of hemoglobin in smokers and non-smokers

More Like This

Show all results sharing this subject:

  • Clinical Cytogenetics and Molecular Genetics


Show Summary Details


Two previous studies demonstrated elevated levels of 3-ethyladenine in smokers' urine, suggesting that cigarette smoke may contain a DNA ethylating agent. We hypothesized that such an agent would also lead to elevated levels of N-terminal N-ethylvaline in hemoglobin. N-terminal N-alkylated valines in hemoglobin can be measured using a modified Edman degradation, which employs pentafluorophenyl isothiocyanate to produce a pentafluorophenylthiohydantoin. The latter is quantified by gas chromatography-negative ion chemical ionization-mass spectrometry (GC-NICI-MS). We modified the published method to increase its sensitivity and selectivity, thereby allowing quantification of N-terminal N-ethylvaline. Modifications included the use of a deuterated peptide as the internal standard, the introduction of an HPLC purification step, and the use of tandem mass spectrometry (MS/MS) for detection and quantification of the analyte, 1-ethyl-5-isopropyl-3-pentafluorophenyl-2-thiohydantoin. We also quantified N-terminal N-methylvaline in the same samples. The mean level of N-terminal N-ethylvaline in the hemoglobin of smokers was 3.76 ± 2.77 pmol/g globin (n = 39), significantly higher than in non-smokers, 2.50 ± 1.65 pmol/g globin (n = 28), P = 0.023. The difference remained significant after correction for gender and age. The mean level of N-terminal N-methylvaline in smokers was 997 ± 203 pmol/g globin (n = 45) compared with 904 ± 149 pmol/g globin in non-smokers (n = 29); these values were not significantly different when corrected for gender and age. As levels of hemoglobin and DNA adducts often correlate, the results of this study support the proposal that cigarette smoke contains an as yet unidentified ethylating agent, which might be involved in DNA damage and tumor initiation.

Keywords: GC-NICI-MS/MS, gas chromatography-negative ion chemical ionization-tandem mass spectrometry; GC-NICI-MS-SIM, gas chromatography-negative ion chemical ionization-mass spectrometry with selected ion monitoring; N-ethylvaline-PFPTH, 1-ethyl-5-isopropyl-3-pentafluorophenyl-2-thiohydantoin; N-methylvaline-PFPTH, 1-methyl-5-isopropyl-3-pentafluorophenyl-2-thiohydantoin; NNK, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone; PAH, polycyclic aromatic hydrocarbons.

Journal Article.  6035 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.