title: Analysis of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB)-releasing DNA adducts in human exfoliated oral mucosa cells by liquid chromatography-electrospray ionization-tandem mass spectrometry.
abstract: Quantitation of DNA adducts could provide critical information on the relationship between exposure to tobacco smoke and cancer risk in smokers. In this study, we developed a robust and sensitive liquid chromatography-tandem mass spectrometry method for the analysis of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB)-releasing DNA adducts in human oral cells, a noninvasive source of DNA for biomarker studies. Isolated DNA undergoes acid hydrolysis, after which samples are purified by solid-phase extraction and analyzed by LC-ESI-MS/MS. The developed method was applied to the analysis of samples obtained via collection with a commercial mouthwash from 30 smokers and 15 nonsmokers. In smokers, the levels of HPB-releasing DNA adducts averaged 12.0 pmol HPB/mg DNA (detected in 20 out of 28 samples with quantifiable DNA yield), and in nonsmokers, the levels of adducts averaged 0.23 pmol/mg DNA (detected in 3 out of 15 samples). For the 30 smoking subjects, matching buccal brushings were also analyzed, and HPB-releasing DNA adducts were detected in 24 out of 27 samples with quantifiable DNA yield, averaging 44.7 pmol HPB/mg DNA. The levels of adducts in buccal brushings correlated with those in mouthwash samples of smokers (R = 0.73, p < 0.0001). Potentially, the method can be applied in studies of individual susceptibility to tobacco-induced cancers in humans.
authors.affiliation: Masonic Cancer Center and ‡Division of Environmental Health Sciences, University of Minnesota , Mayo Mail Code 806, 420 Delaware Street SE, Minneapolis, Minnesota 55455, United States.
authors.forename: Irina
authors.initials: I
authors.lastname: Stepanov
authors.forename: John
authors.initials: J
authors.lastname: Muzic
authors.forename: Chap T
authors.initials: CT
authors.lastname: Le
authors.forename: Erin
authors.initials: E
authors.lastname: Sebero
authors.forename: Peter
authors.initials: P
authors.lastname: Villalta
authors.forename: Bin
authors.initials: B
authors.lastname: Ma
authors.forename: Joni
authors.initials: J
authors.lastname: Jensen
authors.forename: Dorothy
authors.initials: D
authors.lastname: Hatsukami
authors.forename: Stephen S
authors.initials: SS
authors.lastname: Hecht
mesh_terms: D015415:Biomarkers / Q000652:urine; D002074:Butanones / Q000737:chemistry* / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D018736:DNA Adducts / Q000032:analysis* / Q000302:isolation & purification; D006801:Humans; D009061:Mouth Mucosa / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D009067:Mouthwashes / Q000737:chemistry; D009602:Nitrosamines / Q000737:chemistry / Q000378:metabolism; D011725:Pyridines / Q000737:chemistry* / Q000633:toxicity; D012907:Smoking; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D015415:Biomarkers; D002074:Butanones; D018736:DNA Adducts; D009067:Mouthwashes; D009602:Nitrosamines; D011725:Pyridines; C016583:4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone; D004247:DNA; C482281:4-hydroxy-1-(3-pyridyl)-1-butanone