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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| Format: | Article |
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MDPI AG
2021-10-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/21/6355 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-10T05:56:39Z |
| format | Article |
| id | doaj.art-38246f7c79d24838b9bb279ef3126114 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T05:56:39Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-38246f7c79d24838b9bb279ef31261142023-11-22T21:20:03ZengMDPI AGMolecules1420-30492021-10-012621635510.3390/molecules26216355Design and Functionalization of a µPAD for the Enzymatic Determination of Nitrate in UrineFrancisca T. S. M. Ferreira0Raquel B. R. Mesquita1António O. S. S. Rangel2CBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalCBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalCBQF—Centro de Biotecnologia e Química Fina—Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Diogo Botelho 1327, 4169-005 Porto, PortugalIn 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.https://www.mdpi.com/1420-3049/26/21/6355microfluidic paper-based devicenitrate reductasehydrophilic membraneGriess reactionurine samples |
| spellingShingle | 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 Molecules microfluidic paper-based device nitrate reductase hydrophilic membrane Griess reaction urine samples |
| 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 | microfluidic paper-based device nitrate reductase hydrophilic membrane Griess reaction urine samples |
| url | https://www.mdpi.com/1420-3049/26/21/6355 |
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