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Determination of potential childhood asthma biomarkers using a powerful methodology based on microextraction by packed sorbent combined with ultra-high pressure liquid chromatography. Eicosanoids as case study

dc.contributor.authorBerenguer, Pedro
dc.contributor.authorCamacho, Irene C.
dc.contributor.authorCâmara, Rita
dc.contributor.authorOliveira, Susana
dc.contributor.authorCâmara, José S.
dc.date.accessioned2019-12-30T11:12:11Z
dc.date.available2019-12-30T11:12:11Z
dc.date.issued2019
dc.description.abstractLeukotrienes and prostaglandins are arachidonic acid bioactive derived eicosanoids and key mediators of bronchial inflammation and response modulation in the airways contributing to the pathophysiology of asthma. An easy-to-use ultra-high pressure liquid chromatography (UHPLC)-based strategy was developed to characterize biomarkers of lipid peroxidation: leukotrienes E (LTE4) and B4 (LTB4) and 11β-prostaglandin F2α (11βPGF2α), present in urine of asthmatic patients (N = 27) and healthy individuals (N = 17). A semi-automatic eVol®-microextraction by packed sorbent (MEPS) was used to isolate the target analytes. Several experimental parameters with influence on the extraction efficiency and on the chromatographic resolution, were evaluated and optimized. The method was fully validated under optimal extraction (R-AX sorbent, 3 conditioning-equilibration cycles with 250 μL of ACN-water at 0.1% FA, 10 extract-discard cycles of 250 μL of sample at a pH of 5.1, elution with 2 times 50 μL of MeOH and concentration of the eluate until half of its volume) and chromatographic conditions (14-min analysis at a flow rate of 300 μL min-1 in an UHPLC-PDA equipped with a BEH C18 column), according to IUPAC guidelines. The findings indicated good recoveries (>95%) in addition to excellent extraction efficiency (>95%) at three concentration levels (low mid and high) with precision (RSDs) less than 11%. The lack-of-fit test, goodness-of-fit test and Mandel's fitting test, revealed good linearity within the concentration range. Good selectivity and sensitivity were achieved with a limits of detection ranging from 0.04 μg L-1 for LTB4 to 1.12 μg L-1 for 11βPGF2α, and limits of quantification from 0.10 μg L-1 for the LTB4 to 2.11 μg L-1 for 11βPGF2α. The successful application of the fully validated method shows that, on average, the asthmatic patients had significantly higher concentrations of 11βPGF2α (112.96 μg L-1vs 62.56 μg L-1 in normal controls), LTE4 (1.27 μg L-1vs 0.89 μg L-1 in normal controls), and LTB4 (1.39 μg L-1vs 0.76 μg L-1 in normal controls). The results suggest the potential of the target eicosanoids on asthma diagnosis, however, a larger and more extensive study will be necessary to confirm the data obtained and to guarantee a greater robustness to the approach.pt_PT
dc.description.versioninfo:eu-repo/semantics/acceptedVersionpt_PT
dc.identifier.citationhttps://doi.org/10.1016/j.chroma.2018.11.041pt_PT
dc.identifier.doi10.1016/j.chroma.2018.11.041pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.26/30674
dc.language.isoengpt_PT
dc.subjectAsthmapt_PT
dc.subjectBiomarkerspt_PT
dc.subjectCase-Control Studiespt_PT
dc.subjectChildpt_PT
dc.subjectChromatography, High Pressure Liquidpt_PT
dc.subjectEicosanoidspt_PT
dc.subjectFemalept_PT
dc.subjectHumanspt_PT
dc.subjectLimit of Detectionpt_PT
dc.subjectMalept_PT
dc.subjectSolid Phase Microextractionpt_PT
dc.titleDetermination of potential childhood asthma biomarkers using a powerful methodology based on microextraction by packed sorbent combined with ultra-high pressure liquid chromatography. Eicosanoids as case studypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage56pt_PT
oaire.citation.startPage42pt_PT
oaire.citation.titleJournal of Chromatography Apt_PT
oaire.citation.volume1584pt_PT
person.familyNameBerenguer
person.givenNamePedro Henrique Fernandes da Silva
person.identifierD-8897-2019
person.identifier.ciencia-id8619-402B-F57F
person.identifier.orcid0000-0001-8305-2938
person.identifier.scopus-author-id57189574951
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationf8c6d1c8-2259-4322-8d76-464f2d651c33
relation.isAuthorOfPublication.latestForDiscoveryf8c6d1c8-2259-4322-8d76-464f2d651c33

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