Determination of 8-OHGua by LC-MS/MS after Acid Hydrolysis of Oxidative Damaged DNA

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dc.authoridCelik, Saffet/0000-0002-4225-2471
dc.authorwosidCelik, Saffet/R-9630-2017
dc.contributor.authorDanjolli-Hashani, Dua
dc.contributor.authorCelik, Saffet
dc.contributor.authorSelen-Isbilir, Sebnem
dc.date.accessioned2024-06-12T10:50:13Z
dc.date.available2024-06-12T10:50:13Z
dc.date.issued2023
dc.departmentTrakya Üniversitesien_US
dc.description.abstractA variety of influences can cause DNA damage to the genome. The hydroxyl radical attacks the C-8 atom of guanine, forming 8-hydroxyguanine (8-OHGua) or 8-hydroxy-2'-deoxyguanosine (8-OH-2'dG) which are important indicators of oxidative damage on DNA. Determining these damaged base products can be accomplished through measurement by LC-MS/MS after enzymatic hydrolysis or measurement by GC-MS/MS after chemical hydrolysis. In this study, it was aimed to hydrolyze DNA using various strong acids and to measure 8-hydroxyguanine by LC-MS/MS. In the first stage of the study, the nucleoside 8- hydroxy-2'-deoxyguanosine was treated with HCl (2 M and 6 M), TCA (10%), TFA (10%), o-phosphoric acid (2 M), methanesulfonic acid (2 M), and formic acid (80%). The amounts of 8-hydroxyguanine were determined by LC-MS/MS. It has been identified that formic acid with the highest yield (70%) hydrolyzes the beta-glycosidic linkage between the base and the sugar. Subsequently, oxidative damage was induced on calf thymus DNA by producing hydroxyl radicals via the Fenton reaction. The resulting oxidative damaged DNA was hydrolyzed using formic acid. The amount of 8-hydroxyguanine was then determined using LC-MS/MS. Based on the results obtained, it was observed that the acidic hydrolysis applied was effective in breaking the N-glycosidic bond, but not effective in breaking the phosphodiester bond of oxidatively damaged DNA.en_US
dc.description.sponsorshipTrakya University Scientific Research Fund [TUBAP-2019/159]en_US
dc.description.sponsorshipThis study was financially supported by Trakya University Scientific Research Fund (TUBAP-2019/159) . The authors thank the Technology Research Development Application and Research Center (TUTAGEM) for the use of LC-MS/MS.en_US
dc.identifier.endpage409en_US
dc.identifier.issn2383-093X
dc.identifier.issue4en_US
dc.identifier.scopus2-s2.0-85160285283en_US
dc.identifier.scopusqualityQ3en_US
dc.identifier.startpage403en_US
dc.identifier.urihttps://hdl.handle.net/20.500.14551/17915
dc.identifier.volume10en_US
dc.identifier.wosWOS:000996090600002en_US
dc.identifier.wosqualityN/Aen_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.language.isoenen_US
dc.publisherIranian Chemical Socen_US
dc.relation.ispartofAnalytical And Bioanalytical Chemistry Researchen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subject8-OHGen_US
dc.subject8-OH-2'dgen_US
dc.subjectLC-MS/MSen_US
dc.subjectAcidic Hydrolysisen_US
dc.subjectThymus DNAen_US
dc.subjectN-Glycosidic Bonden_US
dc.subjectMass-Spectrometryen_US
dc.subjectRepairen_US
dc.subjectBaseen_US
dc.subject8-Oxo-7,8-Dihydroguanineen_US
dc.subject8-Hydroxydeoxyguanosineen_US
dc.subject8-Oxoguanineen_US
dc.subjectBiomarkersen_US
dc.subjectProtectionen_US
dc.subjectCellsen_US
dc.titleDetermination of 8-OHGua by LC-MS/MS after Acid Hydrolysis of Oxidative Damaged DNAen_US
dc.typeArticleen_US

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