Antioxidants (Jun 2022)

Identification and Quantification of Urinary Microbial Phenolic Metabolites by HPLC-ESI-LTQ-Orbitrap-HRMS and Their Relationship with Dietary Polyphenols in Adolescents

  • Emily P. Laveriano-Santos,
  • María Marhuenda-Muñoz,
  • Anna Vallverdú-Queralt,
  • Miriam Martínez-Huélamo,
  • Anna Tresserra-Rimbau,
  • Elefterios Miliarakis,
  • Camila Arancibia-Riveros,
  • Olga Jáuregui,
  • Ana María Ruiz-León,
  • Sara Castro-Baquero,
  • Ramón Estruch,
  • Patricia Bodega,
  • Mercedes de Miguel,
  • Amaya de Cos-Gandoy,
  • Jesús Martínez-Gómez,
  • Gloria Santos-Beneit,
  • Juan M. Fernández-Alvira,
  • Rodrigo Fernández-Jiménez,
  • Rosa M. Lamuela-Raventós

DOI
https://doi.org/10.3390/antiox11061167
Journal volume & issue
Vol. 11, no. 6
p. 1167

Abstract

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This study aimed to develop and validate a liquid chromatography/electrospray ionization-linear ion trap quadrupole-Orbitrap-high-resolution mass spectrometry (HPLC/ESI-LTQ-Orbitrap-HRMS) method to identify and quantify urinary microbial phenolic metabolites (MPM), as well as to explore the relationship between MPM and dietary (poly)phenols in Spanish adolescents. A total of 601 spot urine samples of adolescents aged 12.02 ± 0.41 years were analyzed. The quantitative method was validated for linearity, limit of detection, limit of quantification, recovery, intra- and inter-day accuracy and precision, as well as postpreparative stability according to the criteria established by the Association of Official Agricultural Chemists International. A total of 17 aglycones and 37 phase II MPM were identified and quantified in 601 spot urine samples. Phenolic acids were the most abundant urinary MPM, whereas stilbenes, hydroxytyrosol, and enterodiol were the least abundant. Urinary hydroxycoumarin acids (urolithins) were positively correlated with flavonoid and total (poly)phenol intake. An HPLC-ESI-LTQ-Orbitrap-HRMS method was developed and fully validated to quantify MPM. The new method was performed accurately and is suitable for MPM quantification in large epidemiological studies. Urinary lignans and urolithins are proposed as potential biomarkers of grain and nut intake in an adolescent population.

Keywords