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Colorimetric determination of urea using diacetyl monoxime with strong acids

Urea is a byproduct of the urea cycle in metabolism and is excreted through urine and sweat. Ammonia, which is toxic at low levels, is converted to the safe storage form of urea, which represents the largest efflux of nitrogen from many organisms. Urea is an important nitrogen source in agriculture,...

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Autores principales: Langenfeld, Noah James, Payne, Lauren Elizabeth, Bugbee, Bruce
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8575183/
https://www.ncbi.nlm.nih.gov/pubmed/34748601
http://dx.doi.org/10.1371/journal.pone.0259760
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author Langenfeld, Noah James
Payne, Lauren Elizabeth
Bugbee, Bruce
author_facet Langenfeld, Noah James
Payne, Lauren Elizabeth
Bugbee, Bruce
author_sort Langenfeld, Noah James
collection PubMed
description Urea is a byproduct of the urea cycle in metabolism and is excreted through urine and sweat. Ammonia, which is toxic at low levels, is converted to the safe storage form of urea, which represents the largest efflux of nitrogen from many organisms. Urea is an important nitrogen source in agriculture, is added to many industrial products, and is a large component in wastewater. The enzyme urease hydrolyzes urea to ammonia and bicarbonate. This reaction is microbially mediated in soils, hydroponic solutions, and wastewater recycling and is catalyzed in vivo in plants using native urease, making measurement of urea environmentally important. Both direct and indirect methods to measure urea exist. This protocol uses diacetyl monoxime to directly determine the concentration of urea in solution. The protocol provides repeatable results and stable reagents with good color stability and simple measurement techniques for use in any lab with a spectrophotometer. The reaction between diacetyl monoxime and urea in the presence of sulfuric acid, phosphoric acid, thiosemicarbazide, and ferric chloride produces a chromophore with a peak absorbance at 520 nm and a linear relationship between concentration and absorbance from 0.4 to 5.0 mM urea in this protocol. The lack of detectable interferences makes this protocol suitable for the determination of millimolar levels of urea in wastewater streams and hydroponic solutions.
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spelling pubmed-85751832021-11-09 Colorimetric determination of urea using diacetyl monoxime with strong acids Langenfeld, Noah James Payne, Lauren Elizabeth Bugbee, Bruce PLoS One Lab Protocol Urea is a byproduct of the urea cycle in metabolism and is excreted through urine and sweat. Ammonia, which is toxic at low levels, is converted to the safe storage form of urea, which represents the largest efflux of nitrogen from many organisms. Urea is an important nitrogen source in agriculture, is added to many industrial products, and is a large component in wastewater. The enzyme urease hydrolyzes urea to ammonia and bicarbonate. This reaction is microbially mediated in soils, hydroponic solutions, and wastewater recycling and is catalyzed in vivo in plants using native urease, making measurement of urea environmentally important. Both direct and indirect methods to measure urea exist. This protocol uses diacetyl monoxime to directly determine the concentration of urea in solution. The protocol provides repeatable results and stable reagents with good color stability and simple measurement techniques for use in any lab with a spectrophotometer. The reaction between diacetyl monoxime and urea in the presence of sulfuric acid, phosphoric acid, thiosemicarbazide, and ferric chloride produces a chromophore with a peak absorbance at 520 nm and a linear relationship between concentration and absorbance from 0.4 to 5.0 mM urea in this protocol. The lack of detectable interferences makes this protocol suitable for the determination of millimolar levels of urea in wastewater streams and hydroponic solutions. Public Library of Science 2021-11-08 /pmc/articles/PMC8575183/ /pubmed/34748601 http://dx.doi.org/10.1371/journal.pone.0259760 Text en © 2021 Langenfeld et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Lab Protocol
Langenfeld, Noah James
Payne, Lauren Elizabeth
Bugbee, Bruce
Colorimetric determination of urea using diacetyl monoxime with strong acids
title Colorimetric determination of urea using diacetyl monoxime with strong acids
title_full Colorimetric determination of urea using diacetyl monoxime with strong acids
title_fullStr Colorimetric determination of urea using diacetyl monoxime with strong acids
title_full_unstemmed Colorimetric determination of urea using diacetyl monoxime with strong acids
title_short Colorimetric determination of urea using diacetyl monoxime with strong acids
title_sort colorimetric determination of urea using diacetyl monoxime with strong acids
topic Lab Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8575183/
https://www.ncbi.nlm.nih.gov/pubmed/34748601
http://dx.doi.org/10.1371/journal.pone.0259760
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