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Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants

Antioxidants prevent ageing and are usually quantified and screened using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. However, this assay cannot be used for salt-containing samples, such as the cell-free supernatants of marine microorganisms that are aggregated under these conditions. Herein, th...

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Autores principales: Gang, Yehui, Eom, Tae-Yang, Marasinghe, Svini Dileepa, Lee, Youngdeuk, Jo, Eunyoung, Oh, Chulhong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146261/
https://www.ncbi.nlm.nih.gov/pubmed/33947091
http://dx.doi.org/10.3390/md19050256
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author Gang, Yehui
Eom, Tae-Yang
Marasinghe, Svini Dileepa
Lee, Youngdeuk
Jo, Eunyoung
Oh, Chulhong
author_facet Gang, Yehui
Eom, Tae-Yang
Marasinghe, Svini Dileepa
Lee, Youngdeuk
Jo, Eunyoung
Oh, Chulhong
author_sort Gang, Yehui
collection PubMed
description Antioxidants prevent ageing and are usually quantified and screened using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. However, this assay cannot be used for salt-containing samples, such as the cell-free supernatants of marine microorganisms that are aggregated under these conditions. Herein, the DPPH solvent (methanol or ethanol) and its water content were optimized to enable the analysis of salt-containing samples, aggregation was observed for alcohol contents of >70%. The water content of methanol influenced the activities of standard antioxidants but did not significantly affect that of the samples. Based on solution stability considerations, 70% aqueous methanol was chosen as the optimal DPPH solvent. The developed method was successfully applied to the cell-free supernatants of marine bacteria (Pseudoalteromonas rubra and Pseudoalteromonas xiamenensis), revealing their high antioxidant activities. Furthermore, it was concluded that this method would be useful for the screening of marine microorganism–derived antioxidants, which also has numerous potential applications, such as salt-fermented foods.
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spelling pubmed-81462612021-05-26 Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants Gang, Yehui Eom, Tae-Yang Marasinghe, Svini Dileepa Lee, Youngdeuk Jo, Eunyoung Oh, Chulhong Mar Drugs Article Antioxidants prevent ageing and are usually quantified and screened using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. However, this assay cannot be used for salt-containing samples, such as the cell-free supernatants of marine microorganisms that are aggregated under these conditions. Herein, the DPPH solvent (methanol or ethanol) and its water content were optimized to enable the analysis of salt-containing samples, aggregation was observed for alcohol contents of >70%. The water content of methanol influenced the activities of standard antioxidants but did not significantly affect that of the samples. Based on solution stability considerations, 70% aqueous methanol was chosen as the optimal DPPH solvent. The developed method was successfully applied to the cell-free supernatants of marine bacteria (Pseudoalteromonas rubra and Pseudoalteromonas xiamenensis), revealing their high antioxidant activities. Furthermore, it was concluded that this method would be useful for the screening of marine microorganism–derived antioxidants, which also has numerous potential applications, such as salt-fermented foods. MDPI 2021-04-29 /pmc/articles/PMC8146261/ /pubmed/33947091 http://dx.doi.org/10.3390/md19050256 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
Gang, Yehui
Eom, Tae-Yang
Marasinghe, Svini Dileepa
Lee, Youngdeuk
Jo, Eunyoung
Oh, Chulhong
Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants
title Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants
title_full Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants
title_fullStr Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants
title_full_unstemmed Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants
title_short Optimising the DPPH Assay for Cell-Free Marine Microorganism Supernatants
title_sort optimising the dpph assay for cell-free marine microorganism supernatants
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146261/
https://www.ncbi.nlm.nih.gov/pubmed/33947091
http://dx.doi.org/10.3390/md19050256
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