Eileen Carry1, Danyue Zhao2, Ilaria Mogno3, Jeremiah Faith3, Lap Ho4, Tom Villani1, Harna Patel2, Giulio M Pasinetti5, James E Simon6, Qingli Wu1. 1. New Use Agriculture & Natural Plant Products Program, Department of Plant Biology, Rutgers University, New Brunswick, NJ 08901, USA; Department of Medicinal Chemistry, Rutgers University, Piscataway, NJ 08854, USA. 2. New Use Agriculture & Natural Plant Products Program, Department of Plant Biology, Rutgers University, New Brunswick, NJ 08901, USA. 3. Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA. 4. Department of Neurology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA. Electronic address: lap.ho@mssm.edu. 5. Department of Neurology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA; Geriatric Research, Education & Clinical Center, James J. Peters Veterans Affairs Medical Center, Bronx, NY, 10468, USA. 6. New Use Agriculture & Natural Plant Products Program, Department of Plant Biology, Rutgers University, New Brunswick, NJ 08901, USA; Department of Medicinal Chemistry, Rutgers University, Piscataway, NJ 08854, USA. Electronic address: jimsimon@rutgers.edu.
Abstract
Grape-derived products contain a wide array of bioactive phenolic compounds which are of significant interest to consumers and researchers for their multiple health benefits. The majority of bioavailable grape polyphenols, including the most abundant flavan-3-ols, i.e. (+)-catechin and (-)-epicatechin, undergo extensive microbial metabolism in the gut, forming metabolites that can be highly bioavailable and bioactive. To gain a better understanding in microbial metabolism of grape polyphenols and to identify bioactive metabolites, advanced analytical methods are needed to accurately quantitate microbial-derived metabolites, particularly at trace levels, in addition to their precursors. This work describes the development and validation of a high-throughput, sensitive and reproducible GC-QqQ/MS method operated under MRM mode that allowed the identification and quantification of 16 phenolic acid metabolites, along with (+)-catechin and (-)-epicatechin, in flavanol-enriched broth samples anaerobically fermented with human intestinal bacteria. Excellent sensitivity was achieved with low limits of detection and low limits of quantification in the range of 0.24-6.18 ng/mL and 0.480-12.37 ng/mL, respectively. With the exception of hippuric acid, recoveries of most analytes were greater than 85%. The percent accuracies for almost all analytes were within ±23% and precision results were all below 18%. Application of the developed method to in vitro samples fermented with different human gut microbiota revealed distinct variations in the extent of flavanol catabolism, as well as production of bioactive phenolic acid metabolites. These results support that intestinal microbiota have a significant impact on the production of flavanol metabolites. The successful application of the established method demonstrates its applicability and robustness for analysis of grape flavanols and their microbial metabolites in biological samples.
Grape-derived products contain a wide array of bioactive phenolic compounds which are of significant interest to consumers and researchers for their multiple health benefits. The majority of bioavailable grape pan class="Chemical">polyphenols, including the most abundant flavan-3-ols, i.e. (+)-catechin and (-)-epicatechin, undergo extensive microbial metabolism in the gut, forming metabolites that can be highly bioavailable and bioactive. To gain a better understanding in microbial metabolism of grape polyphenols and to identify bioactive metabolites, advanced analytical methods are needed to accurately quantitate microbial-derived metabolites, particularly at trace levels, in addition to their precursors. This work describes the development and validation of a high-throughput, sensitive and reproducible GC-QqQ/MS method operated under MRM mode that allowed the identification and quantification of 16 phenolic acid metabolites, along with (+)-catechin and (-)-epicatechin, in flavanol-enriched broth samples anaerobically fermented with human intestinal bacteria. Excellent sensitivity was achieved with low limits of detection and low limits of quantification in the range of 0.24-6.18 ng/mL and 0.480-12.37 ng/mL, respectively. With the exception of hippuric acid, recoveries of most analytes were greater than 85%. The percent accuracies for almost all analytes were within ±23% and precision results were all below 18%. Application of the developed method to in vitro samples fermented with different human gut microbiota revealed distinct variations in the extent of flavanol catabolism, as well as production of bioactive phenolic acid metabolites. These results support that intestinal microbiota have a significant impact on the production of flavanol metabolites. The successful application of the established method demonstrates its applicability and robustness for analysis of grape flavanols and their microbial metabolites in biological samples.
Authors: Álvaro Fernández-Ochoa; María de la Luz Cádiz-Gurrea; Patricia Fernández-Moreno; Alejandro Rojas-García; David Arráez-Román; Antonio Segura-Carretero Journal: Molecules Date: 2022-01-25 Impact factor: 4.411
Authors: Anallely López-Yerena; Inés Domínguez-López; Anna Vallverdú-Queralt; Maria Pérez; Olga Jáuregui; Elvira Escribano-Ferrer; Rosa M Lamuela-Raventós Journal: Antioxidants (Basel) Date: 2021-05-25