title: In vivo detection of DNA adducts induced by cisplatin using capillary HPLC-ICP-MS and their correlation with genotoxic damage in Drosophila melanogaster.
abstract: The antitumoral effect of cisplatin [cis-diamminodichloroplatinum(II)] in mammals is related to its binding to DNA components. However, there is a lack of specific chemical methods to selectively detect those adducts formed in vivo at low concentrations. In this work, a new sensitive and selective method of determining cisplatin-DNA adducts based on the use of element-selective mass spectrometry is proposed, and the method is then applied to detect cisplatin adducts induced in vivo in somatic cells of Drosophila melanogaster. The bioanalytical strategy proposed here allows the determination of the most important DNA adduct formed between adjacent guanine units of the same DNA strand with cisplatin, and it is based on the coupling of capillary liquid chromatography (cap-LC) to inductively coupled plasma mass spectrometry (ICP-MS). This set-up allows the simultaneous monitoring of the Pt (from the drug) and P (from the DNA components) present in these adducts, once they have been cleaved by enzymatic hydrolysis of the DNA samples. Using this instrumental set-up, the adducts of cisplatin formed in vivo when D. melanogaster flies are exposed to different cisplatin concentrations can be detected and their concentration determined. The results obtained show a direct correlation between the concentration of cisplatin adducts, the induced genotoxic damage (measured as DNA strand breaks using the Comet assay) and the cisplatin concentration. [figure: see text] The work illustrates the complementary use of bioanalytical and biological information to study cisplatin interactions with DNA is vivo at biologically relevant concentrations of the drug.
authors.affiliation: Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, C/ Julián Clavería 8, 33006, Oviedo, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: García Sar
authors.forename: Maria
authors.initials: M
authors.lastname: Montes-Bayón
authors.forename: Leticia
authors.initials: L
authors.lastname: Aguado Ortiz
authors.forename: Elisa
authors.initials: E
authors.lastname: Blanco-González
authors.forename: L María
authors.initials: LM
authors.lastname: Sierra
authors.forename: Alfredo
authors.initials: A
authors.lastname: Sanz-Medel
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D002945:Cisplatin / Q000032:analysis* / Q000737:chemistry*; D020552:Comet Assay; D018736:DNA Adducts / Q000032:analysis* / Q000737:chemistry*; D004331:Drosophila melanogaster / Q000187:drug effects / Q000235:genetics*; D005260:Female; D008297:Male; D013058:Mass Spectrometry / Q000295:instrumentation* / Q000379:methods*; D015394:Molecular Structure; D009153:Mutagens / Q000633:toxicity*
chemical_list: D018736:DNA Adducts; D009153:Mutagens; C057270:cisplatin-DNA adduct; D002945:Cisplatin