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A new and fast methodology to assess oxidative damage in cardiovascular diseases risk development through eVol-MEPS–UHPLC analysis of four urinary biomarkers

2013Journal article Restricted access
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dc.contributor.authorMendes, Berta
dc.contributor.authorSilva, Pedro
dc.contributor.authorMendonça, Isabel
dc.contributor.authorPereira, Jorge
dc.contributor.authorCâmara, José S.
dc.date.accessioned2019-07-18T10:40:58Z
dc.date.available2019-07-18T10:40:58Z
dc.date.issued2013
dc.description.abstractIn this work, a new, fast and reliable methodology using a digitally controlled microextraction by packed sorbent (eVol(®)-MEPS) followed by ultra-high pressure liquid chromatography (UHPLC) analysis with photodiodes (PDA) detection, was developed to establish the urinary profile levels of four putative oxidative stress biomarkers (OSBs) in healthy subjects and patients evidencing cardiovascular diseases (CVDs). This data was used to verify the suitability of the selected OSBs (uric acid-UAc, malondialdehyde-MDA, 5-(hydroxymethyl)uracil-5-HMUra and 8-hydroxy-2'-deoxyguanosine-8-oxodG) as potential biomarkers of CVDs progression. Important parameters affecting the efficiency of the extraction process were optimized, particularly stationary phase selection, pH influence, sample volume, number of extraction cycles and washing and elution volumes. The experimental conditions that allowed the best extraction efficiency, expressed in terms of total area of the target analytes and data reproducibility, includes a 10 times dilution and pH adjustment of the urine samples to 6.0, followed by a gradient elution through the C8 adsorbent with 5 times 50 µL of 0.01% formic acid and 3×50 µL of 20% methanol in 0.01% formic acid. The chromatographic separation of the target analytes was performed with a HSS T3 column (100 mm × 2.1 mm, 1.7 µm in particle size) using 0.01% formic acid 20% methanol at 250 µL min(-1). The methodology was validated in terms of selectivity, linearity, instrumental limit of detection (LOD), method limit of quantification (LOQ), matrix effect, accuracy and precision (intra-and inter-day). Good results were obtained in terms of selectivity and linearity (r(2)>0.9906), as well as the LOD and LOQ, whose values were low, ranging from 0.00005 to 0.72 µg mL(-1) and 0.00023 to 2.31 µg mL(-1), respectively. The recovery results (91.1-123.0%), intra-day (1.0-8.3%), inter-day precision (4.6-6.3%) and the matrix effect (60.1-110.3%) of eVol(®)-MEPS/UHPLC-PDA method were also very satisfactory. Finally, the application of the methodology to the determination of target biomarkers in normal subjects and CVDs patients' revealed that the DNA adducts 5-HMUra and 8-oxodG levels are much more abundant in CVDs patients while no statistic differences were obtain for MDA and UAc. This result points to the importance of 5-HMUra and 8-oxodG as biomarkers of CVDs risk progression and further epidemiological studies are needed to explore the importance of this correlation.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1016/j.talanta.2013.04.064pt_PT
dc.identifier.issn0039-9140
dc.identifier.urihttp://hdl.handle.net/10400.13/2480
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.subjectUrinary metabolomic profilept_PT
dc.subjectOxidative stress biomarkers (OSBs)pt_PT
dc.subjectMicroextraction by packed sorbent (MEPS)pt_PT
dc.subjectUltra high pressure liquid chromatographypt_PT
dc.subjectCardiovascular diseases (CVDs)pt_PT
dc.subject.pt_PT
dc.subjectFaculdade de Ciências Exatas e da Engenhariapt_PT
dc.subjectCentro de Química da Madeira
dc.titleA new and fast methodology to assess oxidative damage in cardiovascular diseases risk development through eVol-MEPS–UHPLC analysis of four urinary biomarkerspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F66177%2F2009/PT
oaire.citation.endPage172pt_PT
oaire.citation.startPage164pt_PT
oaire.citation.titleTalantapt_PT
oaire.citation.volume116pt_PT
oaire.fundingStreamSFRH
person.familyNameAugusto Machado Pereira
person.familyNameCâmara
person.givenNameJorge
person.givenNameJosé
person.identifierG-3003-2013
person.identifier.ciencia-idEC12-EE8F-50E8
person.identifier.ciencia-id481C-08CE-90E5
person.identifier.orcid0000-0003-0316-5348
person.identifier.orcid0000-0003-1965-3151
person.identifier.scopus-author-id10140393000
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
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relation.isAuthorOfPublicatione10d78be-e547-4d25-92b5-06a997ed78da
relation.isAuthorOfPublication.latestForDiscovery2f4d7adf-ab6b-40ab-85d5-73995a784bda
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