Optimized Liquid Chromatography Nanoelectrospray-High-Resolution Tandem Mass Spectrometry Method for the Analysis of 4-Hydroxy-1-(3-pyridyl)-1-butanone-Releasing DNA Adducts in Human Oral Cells.
Metabolic activation of the carcinogenic tobacco-specific N-nitrosamines leads to the formation of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB)-releasing DNA adducts. We recently developed a liquid chromatography (LC)-tandem mass spectrometry (MS/MS) method for the analysis of HPB-releasing DNA adducts in human oral cells. However, given the limited amounts of DNA that can be extracted from oral cells, higher sensitivity and selectivity are required for the reliable analysis of these adducts in future studies. We have developed a new sensitive LC-nanoelectrospray ionization-high-resolution MS/MS method for the analysis of HPB-releasing DNA adducts in oral cells. A new procedure was also developed for guanine analysis by LC-MS/MS. The detection limit of the developed assay is 5 amol, and the limit of quantitation is 0.35 fmol HPB on-column, starting with 50 pg of DNA. The method was tested by analyzing oral samples from 65 smokers, including 30 head and neck squamous cell carcinoma (HNSCC) patients and 35 cancer-free controls. In all smokers, the levels of HPB-releasing DNA adducts averaged 6.22 ± 16.18 pmol/mg DNA, with significant interindividual variation being consistent with previous reports. The median HPB-releasing DNA adduct level was 6.6 times greater for those with HNSCC than for smokers without HNSCC (p = 0.002). The developed highly sensitive and selective method is a valuable tool for future measurement of HPB-releasing DNA adducts in tobacco users, which can potentially provide critical insights for the identification of individuals at risk for cancer.
Masonic Cancer Center, ‡Department of Otolaryngology, Head and Neck Surgery, and §Division of Environmental Health Sciences, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Bin
B
Ma
Masonic Cancer Center, ‡Department of Otolaryngology, Head and Neck Surgery, and §Division of Environmental Health Sciences, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Chris
C
Ruszczak
Masonic Cancer Center, ‡Department of Otolaryngology, Head and Neck Surgery, and §Division of Environmental Health Sciences, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Vipin
V
Jain
Masonic Cancer Center, ‡Department of Otolaryngology, Head and Neck Surgery, and §Division of Environmental Health Sciences, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Samir S
SS
Khariwala
Masonic Cancer Center, ‡Department of Otolaryngology, Head and Neck Surgery, and §Division of Environmental Health Sciences, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Bruce
B
Lindgren
Masonic Cancer Center, ‡Department of Otolaryngology, Head and Neck Surgery, and §Division of Environmental Health Sciences, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Dorothy K
DK
Hatsukami
Masonic Cancer Center, ‡Department of Otolaryngology, Head and Neck Surgery, and §Division of Environmental Health Sciences, University of Minnesota , Minneapolis, Minnesota 55455, United States.
Irina
I
Stepanov
D002074:Butanones / Q000633:toxicity*; D002853:Chromatography, Liquid / Q000379:methods*; D018736:DNA Adducts / Q000378:metabolism*; D006801:Humans; D057230:Limit of Detection; D009055:Mouth / Q000166:cytology / Q000378:metabolism*; D036103:Nanotechnology; D011725:Pyridines / Q000633:toxicity*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D002074:Butanones; D018736:DNA Adducts; D011725:Pyridines; C482281:4-hydroxy-1-(3-pyridyl)-1-butanone