Mass spectrometric analysis of tobacco-specific nitrosamine-DNA adducts in smokers and nonsmokers.
A gas chromatography, negative ion chemical ionization mass spectrometry (GC-NICI-MS) based assay for tobacco-specific nitrosamine adducts of DNA is described. The assay is based on the observation that acid hydrolysis of DNA from animals treated with tobacco-specific nitrosamines releases 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB). HPB and the internal standard [4,4-D2]HPB are derivatized with pentafluorobenzoyl chloride and the resulting HPB-pentafluorobenzoate is purified by high-performance liquid chromatography prior to GC-NICI-MS analysis. DNA from human peripheral lung and tracheobronchial tissue, collected at autopsy, was analyzed for acid-released HPB. The mean HPB level (fmol/mg of DNA) for peripheral lung DNA was 11 +/- 16 (SD, n = 9) for smokers and 0.9 +/- 2.3 (n = 8) for nonsmokers. Mean adduct levels in tracheobronchus were 16 +/- 18 (n = 4) for smokers and 0.9 +/- 1.7 (n = 4) for nonsmokers. These are the first measurements of tobacco-specific nitrosamine-DNA adducts in humans. Further studies comparing the levels of DNA and globin adducts will provide a better understanding of the metabolic activation of tobacco-specific nitrosamines in humans and may provide a more accurate indication of an individual's risk of developing tobacco-related cancer.
Foiles
P G
PG
Division of Chemical Carcinogenesis, American Health Foundation, Valhalla, New York 10595.
Akerkar
S A
SA
Carmella
S G
SG
Kagan
M
M
Stoner
G D
GD
Resau
J H
JH
Hecht
S S
SS
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002845:Chromatography / Q000379:methods; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006868:Hydrolysis; D008297:Male; D051379:Mice; D008875:Middle Aged; D009602:Nitrosamines / Q000032:analysis* / Q000378:metabolism; D012907:Smoking / Q000378:metabolism*; D014316:Tritium
D009602:Nitrosamines; D014316:Tritium; D004247:DNA