Repository logo
 
Publication

A Micro-Extraction Technique Using a New Digitally Controlled Syringe Combined with UHPLC for Assessment of Urinary Biomarkers of Oxidatively Damaged DNA

2013-03Journal article Open access
Simple item page
dc.contributor.authorMendes, Berta
dc.contributor.authorSilva, Pedro
dc.contributor.authorAveiro, Fernando
dc.contributor.authorPereira, Jorge
dc.contributor.authorCâmara, José S.
dc.date.accessioned2021-09-27T17:30:18Z
dc.date.available2021-09-27T17:30:18Z
dc.date.issued2013-03
dc.description.abstractThe formation of reactive oxygen species (ROS) within cells causes damage to biomolecules, including membrane lipids, DNA, proteins and sugars. An important type of oxidative damage is DNA base hydroxylation which leads to the formation of 8-oxo-7,8-dihydro-29-deoxyguanosine (8-oxodG) and 5-hydroxymethyluracil (5-HMUra). Measurement of these bio markers in urine is challenging, due to the low levels of the analytes and the matrix complexity. In order to simultaneously quantify 8-oxodG and 5-HMUra in human urine, a new, reliable and powerful strategy was optimised and validated. It is based on a semi-automatic microextraction by packed sorbent (MEPS) technique, using a new digitally controlled syringe (eVolH ), to enhance the extraction efficiency of the target metabolites, followed by a fast and sensitive ultrahigh pressure liquid chromatography (UHPLC). The optimal methodological conditions involve loading of 250 mL urine sample (1:10 dilution) through a C8 sorbent in a MEPS syringe placed in the semi-automatic eVolH syringe followed by elution using 90 mL of 20% methanol in 0.01% formic acid solution. The obtained extract is directly analysed in the UHPLC system using a binary mobile phase composed of aqueous 0.1% formic acid and methanol in the isocratic elution mode (3.5 min total analysis time). The method was validated in terms of selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), extraction yield, accuracy, precision and matrix effect. Satisfactory results were obtained in terms of linearity (r2 . 0.991) within the established concentration range. The LOD varied from 0.00005 to 0.04 mg mL21 and the LOQ from 0.00023 to 0.13 mg mL21 . The extraction yields were between 80.1 and 82.2 %, while inter-day precision (n = 3 days) varied between 4.9 and 7.7 % and intra-day precision between 1.0 and 8.3 %. This approach presents as main advantages the ability to easily collect and store urine samples for further processing and the high sensitivity, reproducibility, and robustness of eVolH MEPS combined with UHPLC analysis, thus retrieving a fast and reliable assessment of oxidatively damaged DNApt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMendes B, Silva P, Aveiro F, Pereira J, Camara JS (2013) A Micro-Extraction Technique Using a New Digitally Controlled Syringe Combined with UHPLC for Assessment of Urinary Biomarkers of Oxidatively Damaged DNA. PLoS ONE 8(3): e58366. doi:10.1371/journal.pone.0058366pt_PT
dc.identifier.doi10.1371/journal.pone.0058366pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.26/37585
dc.language.isoengpt_PT
dc.publisherSusanne Krauss-Etschmann, Ludwig-Maximilians-University Munich, Germanypt_PT
dc.relationStrategic Project - UI 674 - 2011-2012
dc.relation.publisherversionhttps://plosone.orgpt_PT
dc.subjectMendespt_PT
dc.subjectBertapt_PT
dc.subjectSilvapt_PT
dc.subjectPedropt_PT
dc.subjectAveiropt_PT
dc.subjectFernandopt_PT
dc.subjectPereirapt_PT
dc.subjectJorgept_PT
dc.subjectCamarapt_PT
dc.subjectJosé Spt_PT
dc.titleA Micro-Extraction Technique Using a New Digitally Controlled Syringe Combined with UHPLC for Assessment of Urinary Biomarkers of Oxidatively Damaged DNApt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleStrategic Project - UI 674 - 2011-2012
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/PEst-OE%2FQUI%2FUI0674%2F2011/PT
oaire.citation.issue3pt_PT
oaire.citation.startPagee58366pt_PT
oaire.citation.titlePLoS ONEpt_PT
oaire.citation.volume8pt_PT
oaire.fundingStream6817 - DCRRNI ID
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isProjectOfPublicationb2027ec8-fa21-49b6-9231-5aee7839c216
relation.isProjectOfPublication.latestForDiscoveryb2027ec8-fa21-49b6-9231-5aee7839c216

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
A Micro-Extraction Technique Using a New Digitally.pdf
Size:
531.1 KB
Format:
Adobe Portable Document Format
Download

Collections

SESARAM - H/O - Artigos