Rapid and cost-efficient microplate assay for the accurate quantification of total phenolics in seaweeds
•Optical properties of the FBBB interaction with phloroglucinol were investigated.•FC/FBBB ratios of aqueous and ethanoic seaweed extracts ranged from 1.3 to 3.9.•Reaction FBBB and PG to form tri-azo complex is firstly reported.•FBBB method minimized the detection of non-phenolic reducing interferen...
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| Published in | Food chemistry. Molecular sciences Vol. 6; p. 100166 |
|---|---|
| Main Authors | , |
| Format | Journal Article |
| Language | English |
| Published |
England
Elsevier Ltd
30.07.2023
Elsevier |
| Subjects | |
| Online Access | Get full text |
| ISSN | 2666-5662 2666-5662 |
| DOI | 10.1016/j.fochms.2023.100166 |
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| Abstract | •Optical properties of the FBBB interaction with phloroglucinol were investigated.•FC/FBBB ratios of aqueous and ethanoic seaweed extracts ranged from 1.3 to 3.9.•Reaction FBBB and PG to form tri-azo complex is firstly reported.•FBBB method minimized the detection of non-phenolic reducing interferences.•Up to 48 tests (n = 4) in 1 h (0.24€/test) in aqueous and ethanoic seaweeds extracts.
Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format. |
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| AbstractList | Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2-3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format.Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2-3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format. Brown seaweeds ( ) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R ) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2-3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format. • Optical properties of the FBBB interaction with phloroglucinol were investigated. • FC/FBBB ratios of aqueous and ethanoic seaweed extracts ranged from 1.3 to 3.9. • Reaction FBBB and PG to form tri-azo complex is firstly reported. • FBBB method minimized the detection of non-phenolic reducing interferences. • Up to 48 tests (n = 4) in 1 h (0.24€/test) in aqueous and ethanoic seaweeds extracts. Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format. •Optical properties of the FBBB interaction with phloroglucinol were investigated.•FC/FBBB ratios of aqueous and ethanoic seaweed extracts ranged from 1.3 to 3.9.•Reaction FBBB and PG to form tri-azo complex is firstly reported.•FBBB method minimized the detection of non-phenolic reducing interferences.•Up to 48 tests (n = 4) in 1 h (0.24€/test) in aqueous and ethanoic seaweeds extracts. Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format. Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format. |
| ArticleNumber | 100166 |
| Author | Tuohy, Maria G. Martínez-López, Rosalía |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/36875799$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_3390_antiox13040409 |
| Cites_doi | 10.1111/j.1750-3841.2010.01544.x 10.1007/BF02033661 10.1016/S0308-8146(02)00426-0 10.1007/s10811-020-02246-6 10.1016/j.foodchem.2007.08.081 10.1016/j.foodres.2020.109589 10.1016/j.fct.2010.07.029 10.3390/md18010008 10.1016/j.jff.2012.08.007 10.3390/md20070445 10.1016/S0021-9258(18)88697-5 10.1016/j.indcrop.2015.03.053 10.1007/s00227-017-3115-z 10.1515/BOT.2004.057 10.1007/s10811-014-0293-7 10.1111/ijfs.12038 10.1002/pca.3162 10.1016/j.phytol.2015.09.007 10.1021/acs.jafc.7b01000 10.1039/C3RA23373A 10.1093/ajcn/nqy147 10.1139/Y07-105 10.3390/molecules26010043 10.1007/BF02266965 10.1146/annurev-food-032818-121401 10.3390/biomedicines8090342 10.1016/j.crfs.2021.05.005 10.1016/S1381-5148(02)00091-3 10.1039/D0FO01857K 10.1007/s10811-010-9632-5 10.1590/fst.15619 10.1021/jf103711c 10.1002/pca.1210 10.1016/S0021-9258(18)84277-6 10.1007/s10811-006-9048-4 10.1016/j.ijbiomac.2013.04.035 10.1016/j.foodchem.2010.11.111 10.1080/09670260802578542 10.1111/j.1541-4337.2011.00173.x 10.1016/j.jfca.2012.05.003 10.1016/j.jff.2011.02.007 |
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| Keywords | Algae Folin-Ciocalteu Phloroglucinol FBBB F-D Phenolic compounds F-C Seaweeds Fast Blue BB salt PGE FBBB, fast blue bb salt F-C, Folin-Ciocalteu F-D, Folin-Denis PGE, phloroglucinol equivalents |
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Extracts from Edible Marine Algae publication-title: Food Chemistry doi: 10.1016/j.foodchem.2010.11.111 – volume: 44 start-page: 331 issue: 3 year: 2009 ident: 10.1016/j.fochms.2023.100166_b0160 article-title: Seasonal Variation of Polyphenolics in Ascophyllum Nodosum (Phaeophyceae) publication-title: European Journal of Phycology doi: 10.1080/09670260802578542 – volume: 11 start-page: 148 issue: 2 year: 2012 ident: 10.1016/j.fochms.2023.100166_b0045 article-title: Phenol-Based Antioxidants and the In Vitro Methods Used for Their Assessment publication-title: Comprehensive Reviews in Food Science and Food Safety doi: 10.1111/j.1541-4337.2011.00173.x – volume: 27 start-page: 102 issue: 1 year: 2012 ident: 10.1016/j.fochms.2023.100166_b0120 article-title: Comparative Analysis of Strawberry Total Phenolics via Fast Blue BB vs. Folin-Ciocalteu: Assay Interference by Ascorbic Acid publication-title: Journal of Food Composition and Analysis doi: 10.1016/j.jfca.2012.05.003 – volume: 3 start-page: 79 issue: 2 year: 2011 ident: 10.1016/j.fochms.2023.100166_b0125 article-title: Determination of the Total Phenolics in Juices and Superfruits by a Novel Chemical Method publication-title: Journal of Functional Foods doi: 10.1016/j.jff.2011.02.007 |
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| Snippet | •Optical properties of the FBBB interaction with phloroglucinol were investigated.•FC/FBBB ratios of aqueous and ethanoic seaweed extracts ranged from 1.3 to... Brown seaweeds ( ) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the... Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene).... • Optical properties of the FBBB interaction with phloroglucinol were investigated. • FC/FBBB ratios of aqueous and ethanoic seaweed extracts ranged from 1.3... |
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| Title | Rapid and cost-efficient microplate assay for the accurate quantification of total phenolics in seaweeds |
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