title: An improved liquid chromatography-tandem mass spectrometry method for the quantification of 4-aminobiphenyl DNA adducts in urinary bladder cells and tissues.
abstract: Exposure to 4-aminobiphenyl (4-ABP), an environmental and tobacco smoke carcinogen that targets the bladder urothelium, leads to DNA adduct formation and cancer development [1]. Two major analytical challenges in DNA adduct analysis of human samples have been limited sample availability and the need to reach detection limits approaching the part-per-billion threshold. By operating at nano-flow rates and incorporating a capillary analytical column in addition to an online sample enrichment step, we have developed a sensitive and quantitative HPLC-MS/MS method appropriate for the analysis of such samples. This assay for the deoxyguanosine adduct of 4-ABP (dG-C8-4-ABP) gave mass detection limits of 20amol in 1.25microg of DNA (5 adducts in 10(9) nucleosides) with a linear range of 70amol to 70fmol. 4-ABP-exposed human bladder cells and rat bladder tissue were analyzed in triplicate, and higher dose concentrations led to increased numbers of detected adducts. It was subsequently established that sample requirements could be further reduced to 1microg digestions and the equivalent of 250ng DNA per injection for the detection of low levels of dG-C8-4-ABP in a matrix of exfoliated human urothelial cell DNA. This method is appropriate for the characterization and quantification of DNA adducts in human samples and can lead to a greater understanding of their role in carcinogenesis and also facilitate evaluation of chemopreventive agents.
authors.affiliation: Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, 360 Huntington Ave, Boston, MA 02115, USA.
authors.forename: Kristen L
authors.initials: KL
authors.lastname: Randall
authors.forename: Dayana
authors.initials: D
authors.lastname: Argoti
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Paonessa
authors.forename: Yi
authors.initials: Y
authors.lastname: Ding
authors.forename: Zachary
authors.initials: Z
authors.lastname: Oaks
authors.forename: Yuesheng
authors.initials: Y
authors.lastname: Zhang
authors.forename: Paul
authors.initials: P
authors.lastname: Vouros
mesh_terms: D000611:Aminobiphenyl Compounds / Q000633:toxicity*; D000818:Animals; D002273:Carcinogens / Q000494:pharmacology*; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018736:DNA Adducts / Q000032:analysis* / Q000187:drug effects; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D006801:Humans; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D053719:Tandem Mass Spectrometry / Q000379:methods*; D001743:Urinary Bladder / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D019459:Urothelium / Q000737:chemistry / Q000166:cytology / Q000187:drug effects
chemical_list: D000611:Aminobiphenyl Compounds; D002273:Carcinogens; D018736:DNA Adducts; C006757:4-biphenylamine