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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: Ferreira, Francisca T. S. M., Mesquita, Raquel B. R., Rangel, António O. S. S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8588010/
https://www.ncbi.nlm.nih.gov/pubmed/34770764
http://dx.doi.org/10.3390/molecules26216355
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author Ferreira, Francisca T. S. M.
Mesquita, Raquel B. R.
Rangel, António O. S. S.
author_facet Ferreira, Francisca T. S. M.
Mesquita, Raquel B. R.
Rangel, António O. S. S.
author_sort Ferreira, Francisca T. S. M.
collection PubMed
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.
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spelling pubmed-85880102021-11-13 Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine Ferreira, Francisca T. S. M. Mesquita, Raquel B. R. Rangel, António O. S. S. Molecules Article 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. MDPI 2021-10-21 /pmc/articles/PMC8588010/ /pubmed/34770764 http://dx.doi.org/10.3390/molecules26216355 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ferreira, Francisca T. S. M.
Mesquita, Raquel B. R.
Rangel, António O. S. S.
Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in Urine
title 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_short 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8588010/
https://www.ncbi.nlm.nih.gov/pubmed/34770764
http://dx.doi.org/10.3390/molecules26216355
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