Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine

In this work, the design of a microfluidic paper-based analytical device (μPAD) for the quantification of nitrate in urine samples was described. Nitrate monitoring is highly relevant due to its association to some diseases and health conditions. The nitrate determination was achieved by combining t...

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Autores principales:	Francisca T. S. M. Ferreira, Raquel B. R. Mesquita, António O. S. S. Rangel
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	microfluidic paper-based device nitrate reductase hydrophilic membrane Griess reaction urine samples Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/38246f7c79d24838b9bb279ef3126114
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spelling	oai:doaj.org-article:38246f7c79d24838b9bb279ef31261142021-11-11T18:23:06ZDesign and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine10.3390/molecules262163551420-3049https://doaj.org/article/38246f7c79d24838b9bb279ef31261142021-10-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/21/6355https://doaj.org/toc/1420-3049In this work, the design of a microfluidic paper-based analytical device (μPAD) for the quantification of nitrate in urine samples was described. Nitrate monitoring is highly relevant due to its association to some diseases and health conditions. The nitrate determination was achieved by combining the selectivity of the nitrate reductase enzymatic reaction with the colorimetric detection of nitrite by the well-known Griess reagent. For the optimization of the nitrate determination μPAD, several variables associated with the design and construction of the device were studied. Furthermore, the interference of the urine matrix was evaluated, and stability studies were performed, under different conditions. The developed μPAD enabled us to obtain a limit of detection of 0.04 mM, a limit of quantification of 0.14 mM and a dynamic concentration range of 0.14–1.0 mM. The designed μPAD proved to be stable for 24 h when stored at room temperature in air or vacuum atmosphere, and 60 days when stored in vacuum at −20 °C. The accuracy of the nitrate μPAD measurements was confirmed by analyzing four certified samples (prepared in synthetic urine) and performing recovery studies using urine samples.Francisca T. S. M. FerreiraRaquel B. R. MesquitaAntónio O. S. S. RangelMDPI AGarticlemicrofluidic paper-based devicenitrate reductasehydrophilic membraneGriess reactionurine samplesOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6355, p 6355 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	microfluidic paper-based device nitrate reductase hydrophilic membrane Griess reaction urine samples Organic chemistry QD241-441
spellingShingle	microfluidic paper-based device nitrate reductase hydrophilic membrane Griess reaction urine samples Organic chemistry QD241-441 Francisca T. S. M. Ferreira Raquel B. R. Mesquita António O. S. S. Rangel Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine
description	In this work, the design of a microfluidic paper-based analytical device (μPAD) for the quantification of nitrate in urine samples was described. Nitrate monitoring is highly relevant due to its association to some diseases and health conditions. The nitrate determination was achieved by combining the selectivity of the nitrate reductase enzymatic reaction with the colorimetric detection of nitrite by the well-known Griess reagent. For the optimization of the nitrate determination μPAD, several variables associated with the design and construction of the device were studied. Furthermore, the interference of the urine matrix was evaluated, and stability studies were performed, under different conditions. The developed μPAD enabled us to obtain a limit of detection of 0.04 mM, a limit of quantification of 0.14 mM and a dynamic concentration range of 0.14–1.0 mM. The designed μPAD proved to be stable for 24 h when stored at room temperature in air or vacuum atmosphere, and 60 days when stored in vacuum at −20 °C. The accuracy of the nitrate μPAD measurements was confirmed by analyzing four certified samples (prepared in synthetic urine) and performing recovery studies using urine samples.
format	article
author	Francisca T. S. M. Ferreira Raquel B. R. Mesquita António O. S. S. Rangel
author_facet	Francisca T. S. M. Ferreira Raquel B. R. Mesquita António O. S. S. Rangel
author_sort	Francisca T. S. M. Ferreira
title	Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine
title_short	Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine
title_full	Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine
title_fullStr	Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine
title_full_unstemmed	Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine
title_sort	design and functionalization of a µpad for the enzymatic determination of nitrate in urine
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/38246f7c79d24838b9bb279ef3126114
work_keys_str_mv	AT franciscatsmferreira designandfunctionalizationofaμpadfortheenzymaticdeterminationofnitrateinurine AT raquelbrmesquita designandfunctionalizationofaμpadfortheenzymaticdeterminationofnitrateinurine AT antonioossrangel designandfunctionalizationofaμpadfortheenzymaticdeterminationofnitrateinurine
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Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine

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