Development, optimization and validation of LC-MS/MS method for the determination of DBS GALT enzyme activity

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dc.authorscopusid58535123600
dc.authorscopusid55896960200
dc.authorscopusid6701587100
dc.authorscopusid23059712400
dc.authorscopusid26428601400
dc.authorscopusid7007002166
dc.authorscopusid6506719953
dc.contributor.authorTopbas, M.
dc.contributor.authorCanbay, E.
dc.contributor.authorSezer, E.
dc.contributor.authorCanda, E.
dc.contributor.authorKalkan, Uçar, S.
dc.contributor.authorÇoker, M.
dc.contributor.authorYildirim, Sözmen, E.
dc.date.accessioned2024-08-25T18:31:31Z
dc.date.available2024-08-25T18:31:31Z
dc.date.issued2023
dc.departmentEge Üniversitesien_US
dc.description.abstractGalactosemia is a carbohydrate metabolism disorder often caused by galactose-1-phosphate uridyl transferase (GALT) deficiency. Detecting GALT deficiency involves measuring intra-erythrocyte enzyme activity. We aimed to create a robust liquid chromatography-mass spectrometry (LC-MS/MS) method to assess GALT activity in dried blood spot (DBS) samples. We validated this method and compared it to the fluorometric approach. We investigated the impact of K2EDTA and lithium heparin tubes on enzyme activity to identify the best sample collection tube. We also assessed the reaction-stopping method. The developed approach employed [13C6]-galactose-1-phosphate as a substrate and UDP-N-acetylglycosamine as an internal standard (IS). The mean ± SD value for GALT activity of DBS samples was determined as 6.37 ± 1.96 ?mol/gHb/hour. The linear range was 0.4–50 ?M (2.4–310% of normal) in the DBS method. The % coefficient of variation (%CV) values were less than 15 for intra-day and inter-day repeatability studies. Over 90% recovery was achieved in recovery studies, and no ion suppression from matrix was detected. DBS samples were quite stable for 31 days under different storage conditions. Enzyme activity results reported as <3.5 U/g Hb by fluorometric method, were quantitatively determined for even very low concentrations by LC-MS/MS method. © 2023 Elsevier Inc.en_US
dc.description.sponsorshipEge Üniversitesi: TTU-2019-20812en_US
dc.description.sponsorshipThis project was funded by the Ege University Research Fund (Project Number TTU-2019-20812 ).en_US
dc.identifier.doi10.1016/j.ab.2023.115284
dc.identifier.issn0003-2697
dc.identifier.pmid37572839en_US
dc.identifier.scopus2-s2.0-85167807882en_US
dc.identifier.scopusqualityQ3en_US
dc.identifier.urihttps://doi.org/10.1016/j.ab.2023.115284
dc.identifier.urihttps://hdl.handle.net/11454/99919
dc.identifier.volume678en_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherAcademic Press Inc.en_US
dc.relation.ispartofAnalytical Biochemistryen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.snmz20240825_Gen_US
dc.subjectDried blood spoten_US
dc.subjectGALTen_US
dc.subjectLC-MS/MSen_US
dc.subjectNewborn screeningen_US
dc.subjectBlooden_US
dc.subjectLiquid chromatographyen_US
dc.subjectMass spectrometryen_US
dc.subjectCarbohydrate metabolismen_US
dc.subjectDried blood spoten_US
dc.subjectEnzymes activityen_US
dc.subjectGALTen_US
dc.subjectLC-MS-MSen_US
dc.subjectLc.ms/msen_US
dc.subjectLiquid chromatography - mass spectrometriesen_US
dc.subjectNewborn screeningen_US
dc.subjectOptimisationsen_US
dc.subjectSpot samplesen_US
dc.subjectEnzyme activityen_US
dc.subjectacetonitrileen_US
dc.subjectgalactose 1 phosphate uridylyltransferaseen_US
dc.subjectgalactose 1 phosphate uridylyltransferaseen_US
dc.subjectadolescenten_US
dc.subjectadulten_US
dc.subjectageden_US
dc.subjectArticleen_US
dc.subjectchilden_US
dc.subjectcontrolled studyen_US
dc.subjectdried blood spot testingen_US
dc.subjectenzyme activityen_US
dc.subjectfemaleen_US
dc.subjectfluorometryen_US
dc.subjecthumanen_US
dc.subjectinfanten_US
dc.subjectintermethod comparisonen_US
dc.subjectliquid chromatography-mass spectrometryen_US
dc.subjectmaleen_US
dc.subjectmatrix effecten_US
dc.subjectmeasurement accuracyen_US
dc.subjectmeasurement repeatabilityen_US
dc.subjectnewbornen_US
dc.subjectnormal humanen_US
dc.subjectquantitative analysisen_US
dc.subjectstorageen_US
dc.subjectvalidation processen_US
dc.subjectdried blood spot testingen_US
dc.subjectgalactosemiaen_US
dc.subjectliquid chromatographyen_US
dc.subjectproceduresen_US
dc.subjectreproducibilityen_US
dc.subjecttandem mass spectrometryen_US
dc.subjectChromatography, Liquiden_US
dc.subjectDried Blood Spot Testingen_US
dc.subjectGalactosemiasen_US
dc.subjectHumansen_US
dc.subjectReproducibility of Resultsen_US
dc.subjectTandem Mass Spectrometryen_US
dc.subjectUTP-Hexose-1-Phosphate Uridylyltransferaseen_US
dc.titleDevelopment, optimization and validation of LC-MS/MS method for the determination of DBS GALT enzyme activityen_US
dc.typeArticleen_US

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