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A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume

There is ample evidence that polyphenols are important natural substances with pronounced antioxidative properties. This study aimed to develop a fast and reliable method to determine total polyphenol content (TPC) in foodstuffs and human samples. The microtitration format offers the advantage of lo...

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Autores principales: Tatzber, Franz, Wonisch, Willibald, Lackner, Sonja, Lindschinger, Meinrad, Pursch, Werner, Resch, Ulrike, Trummer, Christopher, Murkovic, Michael, Zelzer, Sieglinde, Holasek, Sandra, Cvirn, Gerhard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7023636/
https://www.ncbi.nlm.nih.gov/pubmed/31877807
http://dx.doi.org/10.3390/antiox9010011
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author Tatzber, Franz
Wonisch, Willibald
Lackner, Sonja
Lindschinger, Meinrad
Pursch, Werner
Resch, Ulrike
Trummer, Christopher
Murkovic, Michael
Zelzer, Sieglinde
Holasek, Sandra
Cvirn, Gerhard
author_facet Tatzber, Franz
Wonisch, Willibald
Lackner, Sonja
Lindschinger, Meinrad
Pursch, Werner
Resch, Ulrike
Trummer, Christopher
Murkovic, Michael
Zelzer, Sieglinde
Holasek, Sandra
Cvirn, Gerhard
author_sort Tatzber, Franz
collection PubMed
description There is ample evidence that polyphenols are important natural substances with pronounced antioxidative properties. This study aimed to develop a fast and reliable method to determine total polyphenol content (TPC) in foodstuffs and human samples. The microtitration format offers the advantage of low sample volumes in the microlitre range, facilitating high-throughput screening with 40 samples simultaneously. We accordingly adjusted the so-called Folin–Ciocalteu method to a microtitre format (polyphenols microtitre—PPm) with 90% reduction of reagents. The assay was standardized with gallic acid in the range between 0.1 and 3 mM, using a 20 µL sample volume. The intra-assay coefficient of variation (CV) was less than 5%, and inter-assay CV was in the range of 10%. Wavelength was measured at 766 nm after two hours of incubation. This micromethod correlates significantly with both the classical Folin–Ciocalteu method and High-Performance Thin-Layer Chromatography (HPTLC) (r(2) = 0.9829). We further observed a significant correlation between PPm and total antioxidants (r(2) = 0.918). The highest polyphenol concentrations were obtained for red, blue, and black fruits, vegetables, and juices. Extracts of red grapes could be harvested almost sugar free and might serve as a basis for polyphenol supplementation. Beer, flour, and bread contained polyphenol concentrations sufficient to meet the minimal daily requirement. We conclude that PPm is a sensitive and reliable method that detects polyphenols even in samples diluted 10-fold. The literature strongly recommends further investigations on the effects of polyphenol uptake on human and animal health.
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spelling pubmed-70236362020-03-11 A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume Tatzber, Franz Wonisch, Willibald Lackner, Sonja Lindschinger, Meinrad Pursch, Werner Resch, Ulrike Trummer, Christopher Murkovic, Michael Zelzer, Sieglinde Holasek, Sandra Cvirn, Gerhard Antioxidants (Basel) Article There is ample evidence that polyphenols are important natural substances with pronounced antioxidative properties. This study aimed to develop a fast and reliable method to determine total polyphenol content (TPC) in foodstuffs and human samples. The microtitration format offers the advantage of low sample volumes in the microlitre range, facilitating high-throughput screening with 40 samples simultaneously. We accordingly adjusted the so-called Folin–Ciocalteu method to a microtitre format (polyphenols microtitre—PPm) with 90% reduction of reagents. The assay was standardized with gallic acid in the range between 0.1 and 3 mM, using a 20 µL sample volume. The intra-assay coefficient of variation (CV) was less than 5%, and inter-assay CV was in the range of 10%. Wavelength was measured at 766 nm after two hours of incubation. This micromethod correlates significantly with both the classical Folin–Ciocalteu method and High-Performance Thin-Layer Chromatography (HPTLC) (r(2) = 0.9829). We further observed a significant correlation between PPm and total antioxidants (r(2) = 0.918). The highest polyphenol concentrations were obtained for red, blue, and black fruits, vegetables, and juices. Extracts of red grapes could be harvested almost sugar free and might serve as a basis for polyphenol supplementation. Beer, flour, and bread contained polyphenol concentrations sufficient to meet the minimal daily requirement. We conclude that PPm is a sensitive and reliable method that detects polyphenols even in samples diluted 10-fold. The literature strongly recommends further investigations on the effects of polyphenol uptake on human and animal health. MDPI 2019-12-21 /pmc/articles/PMC7023636/ /pubmed/31877807 http://dx.doi.org/10.3390/antiox9010011 Text en © 2019 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tatzber, Franz
Wonisch, Willibald
Lackner, Sonja
Lindschinger, Meinrad
Pursch, Werner
Resch, Ulrike
Trummer, Christopher
Murkovic, Michael
Zelzer, Sieglinde
Holasek, Sandra
Cvirn, Gerhard
A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume
title A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume
title_full A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume
title_fullStr A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume
title_full_unstemmed A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume
title_short A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume
title_sort micromethod for polyphenol high-throughput screening saves 90 percent reagents and sample volume
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7023636/
https://www.ncbi.nlm.nih.gov/pubmed/31877807
http://dx.doi.org/10.3390/antiox9010011
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