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Development of a new HPLC-based method for 3-nitrotyrosine quantification in different biological matrices

2017Journal article Open access
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dc.contributor.authorTeixeira, Dulce
dc.contributor.authorPrudêncio, Cristina
dc.contributor.authorVieira, Mónica
dc.date.accessioned2019-07-01T14:37:24Z
dc.date.available2022-05-02T00:30:47Z
dc.date.issued2017
dc.description.abstractThe nitration of tyrosine residues in proteins is associated with nitrosative stress, resulting in the formation of 3-nitrotyrosine (3-NT).1 3-NT levels in biological samples have been associated with numerous physiological and pathological conditions. Hence several attempts have been made in order to develop methods that accurately quantify 3-NT in these matrices.The aim of this study was to develop a simple, rapid, low-cost and sensitive high-performance liquid chromatography (HPLC)-based 3-NT quantification method. Methods All experiments were performed on an Hitachi LaChrom Elite® HPLC system. The method was validated according to International Conference on Harmonisation (ICH) guidelines for serum samples. Additionally, other biological matrices were tested, namely whole blood, urine, B16 F-10 melanoma cell line, growth medium conditioned with the same cell line, bacterial and yeast suspensions. Results From all the protocols tested, the best results were obtained using 0.5% CH3COOH:MeOH:H2O (15:15:70) as mobile phase, with detection at wavelengths 215, 276 and 356 nm, at 25 °C, and using a flow rate of 1 mL min−1. By using this protocol, it was possible to obtain a linear calibration curve, limits of detection and quantification in the order of μg L−1, and a short analysis time (<15 min per sample). The developed protocol allowed the successful detection and quantification of 3-NT in all biological matrices tested, with detection at 356 nm. Conclusion This method, successfully developed and validated for 3-NT quantification, is simple, cheap and fast. These features render this method a suitable option for analysis of a wide range of biological matrices, being a promising useful tool for both research and diagnosis activities.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationTeixeira, D., Prudêncio, C., & Vieira, M. (2017). Development of a new HPLC-based method for 3-nitrotyrosine quantification in different biological matrices. Journal of Chromatography B, 1046, 48–57. https://doi.org/10.1016/j.jchromb.2017.01.035
dc.identifier.doi10.1016/j.jchromb.2017.01.035pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.22/14195
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S1570023216308893pt_PT
dc.subject3-Nitrotyrosinept_PT
dc.subjectNitrosative stresspt_PT
dc.subjectHPLC-DADpt_PT
dc.subjectQuantification methodspt_PT
dc.titleDevelopment of a new HPLC-based method for 3-nitrotyrosine quantification in different biological matricespt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage57pt_PT
oaire.citation.startPage48pt_PT
oaire.citation.titleJournal of Chromatography Bpt_PT
oaire.citation.volume1046pt_PT
person.familyNameTeixeira
person.familyNamePrudêncio
person.familyNameVieira
person.givenNameDulce
person.givenNameCristina
person.givenNameMónica
person.identifier1200571
person.identifier.ciencia-idF217-0942-4461
person.identifier.ciencia-idC81E-F4EE-FADE
person.identifier.orcid0000-0002-9388-9018
person.identifier.orcid0000-0002-9920-936X
person.identifier.orcid0000-0002-9614-9031
person.identifier.ridD-8977-2016
person.identifier.scopus-author-id6508057930
person.identifier.scopus-author-id24464652500
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationc7338403-0b6f-48fb-ac57-30d0e652496d
relation.isAuthorOfPublication881a8ad5-ab13-4e49-89f4-08ca61cc81e3
relation.isAuthorOfPublication55587c4b-3df0-4ff1-bade-587420940ecd
relation.isAuthorOfPublication.latestForDiscoveryc7338403-0b6f-48fb-ac57-30d0e652496d

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