title: Adduct levels from benzo[a]pyrenediol epoxide: Relative formation to histidine in serum albumin and to deoxyguanosine in DNA in vitro and in vivo in mice measured by LC/MS-MS methods.
abstract: Stable and specific biomacromolecular adducts can be used to measure in vivo doses of reactive compounds. An LC/MS-MS method to measure adducts from the benzo[a]pyrene (BP) metabolite (±)-anti-BP-7,8-diol-9,10-epoxide ((±)-anti-BPDE) to His(146) in serum albumin (SA), earlier evaluated on in vitro alkylated human SA, was tested for its applicability to mouse. It was shown that (+)-anti-BPDE form BPDE-His adducts to mouse SA. The method was applied to samples from BP-exposed mice (100mg/kg of body weight for 1, 3, 7 and 28 days). BPDE-His in SA was close to the limit of quantification and showed the highest level (13fmol/mg) 3 days after exposure. The level was 400 times lower (calculated per gram macromolecule) than earlier measured level of BPDE-adduct to deoxyguanosine (dG) in DNA in the livers. The relative rate of formation of adducts from BPDE with His in SA and with dG in DNA was investigated. Quantification by LC/MS-MS of the adducts in human blood alkylated in vitro with (±)-anti-BPDE showed a 1850 times higher level of BPDE-dG compared to BPDE-His. The specific and stable BPDE-adducts to His in SA are potential biomarkers of in vivo dose of BPDE, though this requires a considerable improved analytical sensitivity of the LC/MS-MS method.
authors.affiliation: Division of Environmental Chemistry, Department of Materials and Environmental Chemistry, Stockholm University, SE-10691 Stockholm, Sweden. Electronic address: emelie.westberg@mmk.su.se.
authors.forename: Emelie A C
authors.initials: EA
authors.lastname: Westberg
authors.affiliation: Department of Cancer Studies & Molecular Medicine, Robert Kilpatrick Clinical Sciences Building, University of Leicester, Leicester Royal Infirmary, Leicester LE27LX, UK. Electronic address: rs25@leicester.ac.uk.
authors.forename: Rajinder
authors.initials: R
authors.lastname: Singh
authors.affiliation: Division of Environmental Chemistry, Department of Materials and Environmental Chemistry, Stockholm University, SE-10691 Stockholm, Sweden. Electronic address: ulla.hedebrant@gmail.com.
authors.forename: Ulla
authors.initials: U
authors.lastname: Hedebrant
authors.affiliation: Institute of Biology, Pharmaceutical Chemistry and Biotechnology, National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, Athens 11635, Greece.
authors.forename: Georgios
authors.initials: G
authors.lastname: Koukouves
authors.affiliation: Institute of Biology, Pharmaceutical Chemistry and Biotechnology, National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, Athens 11635, Greece. Electronic address: vls@eie.gr.
authors.forename: Vassilis L
authors.initials: VL
authors.lastname: Souliotis
authors.affiliation: Department of Cancer Studies & Molecular Medicine, Robert Kilpatrick Clinical Sciences Building, University of Leicester, Leicester Royal Infirmary, Leicester LE27LX, UK. Electronic address: pbf1@le.ac.uk.
authors.forename: Peter B
authors.initials: PB
authors.lastname: Farmer
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institute, Novum, SE-14183 Huddinge, Sweden. Electronic address: dan.segerback@ki.se.
authors.forename: Dan
authors.initials: D
authors.lastname: Segerbäck
authors.affiliation: Institute of Biology, Pharmaceutical Chemistry and Biotechnology, National Hellenic Research Foundation, 48 Vassileos Constantinou Avenue, Athens 11635, Greece. Electronic address: skyrt@eie.gr.
authors.forename: Soterios
authors.initials: S
authors.lastname: Kyrtopoulos
authors.affiliation: Division of Environmental Chemistry, Department of Materials and Environmental Chemistry, Stockholm University, SE-10691 Stockholm, Sweden. Electronic address: margareta.tornqvist@mmk.su.se.
authors.forename: Margareta Å
authors.initials: MÅ
authors.lastname: Törnqvist
mesh_terms: D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000378:metabolism* / Q000633:toxicity; D000818:Animals; D001564:Benzo(a)pyrene; D015415:Biomarkers / Q000097:blood; D002853:Chromatography, Liquid; D018736:DNA Adducts / Q000097:blood*; D003849:Deoxyguanosine; D006639:Histidine; D006801:Humans; D008297:Male; D051379:Mice; D011485:Protein Binding; D018570:Risk Assessment; D012709:Serum Albumin / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D015415:Biomarkers; D018736:DNA Adducts; D012709:Serum Albumin; C056612:benzo(a)pyrene-DNA adduct; D001564:Benzo(a)pyrene; D006639:Histidine; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D003849:Deoxyguanosine
keywords: Benzo[a]pyrene diolepoxide; DNA adduct; HPLC/MS–MS; Histidine adduct; Serum albumin