title: Detection of DNA adducts originating from 1-methoxy-3-indolylmethyl glucosinolate using isotope-dilution UPLC-ESI-MS/MS.
abstract: 1-Methoxy-3-indolylmethyl (1-MIM) glucosinolate, present at substantial levels in several food crops (e.g., broccoli and cabbage), forms DNA adducts in vitro and is mutagenic to bacterial and mammalian cells after activation by the plant enzyme myrosinase. Moreover, a breakdown product, 1-MIM alcohol, is metabolized to a secondary reactive intermediate by some mammalian sulfotransferases (SULTs). First, we incubated herring-sperm DNA with 1-MIM glucosinolate in the presence of myrosinase. We identified and synthesized the predominant adducts, N(2)-(1-MIM)-dG and N(6)-(1-MIM)-dA, and developed an UPLC-ESI-MS/MS method for their specific detection using isotopic dilution. Second, we demonstrated both DNA adducts in target cells (Salmonella typhimurium TA100 and Chinese hamster V79) of standard mutagenicity tests treated with 1-MIM glucosinolate/myrosinase as well as in 1-MIM alcohol-treated Salmonella and V79 cells engineered for expression of human SULT1A1. Similar excesses of N(2)-(1-MIM)-dG over N(6)-(1-MIM)-dA adducts were found in all cellular models independent of the test compound (1-MIM glucosinolate or alcohol), whereas dA adducts predominated in the cell-free system. Finally, we detected both DNA adducts in colon tissue of a mouse orally treated with 1-MIM glucosinolate. We are going to use this specific and sensitive method for investigating genotoxic risks of food-borne exposure to 1-MIM glucosinolate in animal and human studies.
authors: German Institute of Human Nutrition, Potsdam-Rehbrücke, 14558 Nuthetal, Germany.
authors: Fabian
authors: F
authors: Schumacher
authors: Wolfram
authors: W
authors: Engst
authors: Bernhard H
authors: BH
authors: Monien
authors: Simone
authors: S
authors: Florian
authors: Anke
authors: A
authors: Schnapper
authors: Lisa
authors: L
authors: Steinhauser
authors: Klaus
authors: K
authors: Albert
authors: Heinz
authors: H
authors: Frank
authors: Albrecht
authors: A
authors: Seidel
authors: Hansruedi
authors: H
authors: Glatt
mesh_terms: D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D018736:DNA Adducts / Q000378:metabolism*; D005399:Fishes; D005961:Glucosinolates / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007211:Indoles / Q000378:metabolism*; D007554:Isotopes; D057230:Limit of Detection; D008297:Male; D051379:Mice; D012486:Salmonella typhimurium / Q000166:cytology; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013094:Spermatozoa / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D018736:DNA Adducts; D005961:Glucosinolates; D007211:Indoles; D007554:Isotopes; C064701:1-methoxy-3-indolylmethylglucosinolate