TY - JOUR
T1 - Targeted analysis of microbial-generated phenolic acid metabolites derived from grape flavanols by gas chromatography-triple quadrupole mass spectrometry
AU - Carry, Eileen
AU - Zhao, Danyue
AU - Mogno, Ilaria
AU - Faith, Jeremiah
AU - Ho, Lap
AU - Villani, Tom
AU - Patel, Harna
AU - Pasinetti, Giulio M.
AU - Simon, James E.
AU - Wu, Qingli
N1 - Funding Information:
This work was done as part of the Core B research program in NIH Botanical Center with funds provided by NIH ODS and NCCAM IP50AT008661-01 to Mt. Sinai and with Supplemental Funding by the NIH to Mt. Sinai (Grant number 0254-3831-4609 ). We thank Shimadzu Instruments for the analytical instruments used to conduct this research. Funds were also provided by the New Jersey Agricultural Experiment Station Hatch Project Number NJ12158. The authors are very thankful to Dr. Mario Ferruzzi in North Carolina State University and Dr. Rick Dixon, University of North Texas for their constructive suggestions.
Funding Information:
This work was done as part of the Core B research program in NIH Botanical Center with funds provided by NIH ODS and NCCAM IP50AT008661-01 to Mt. Sinai and with Supplemental Funding by the NIH to Mt. Sinai (Grant number 0254-3831-4609). We thank Shimadzu Instruments for the analytical instruments used to conduct this research. Funds were also provided by the New Jersey Agricultural Experiment Station Hatch Project Number NJ12158. The authors are very thankful to Dr. Mario Ferruzzi in North Carolina State University and Dr. Rick Dixon, University of North Texas for their constructive suggestions.
Publisher Copyright:
© 2018
PY - 2018/9/10
Y1 - 2018/9/10
N2 - 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.
AB - 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.
KW - GC-QqQ/MS
KW - Grape polyphenol
KW - Gut microbiota in vitro fermentation
KW - Phenolic acid metabolite
UR - http://www.scopus.com/inward/record.url?scp=85050088099&partnerID=8YFLogxK
U2 - 10.1016/j.jpba.2018.06.034
DO - 10.1016/j.jpba.2018.06.034
M3 - Article
C2 - 30032004
AN - SCOPUS:85050088099
SN - 0731-7085
VL - 159
SP - 374
EP - 383
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
ER -