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Urinary metabolomic profiling to identify biomarkers of a flavonoid-rich and flavonoid-poor fruits and vegetables diet in adults: the FLAVURS trial

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Abstract

The present study aims to investigate the dose dependent effects of consuming diets enriched in flavonoid-rich and flavonoid-poor fruits and vegetables on the urine metabolome of adults who had a ≥1.5 fold increased risk of cardiovascular diseases. A single-blind, dose-dependent, parallel randomized controlled dietary intervention was conducted where volunteers (n = 126) were randomly assigned to one of three diets: high flavonoid diet, low flavonoid diet or habitual diet as a control for 18 weeks. High resolution LC–MS untargeted metabolomics with minimal sample cleanup was performed using an Orbitrap mass spectrometer. Putative biomarkers which characterize diets with high and low flavonoid content were selected by state-of-the-art data analysis strategies and identified by HR-MS and HR-MS/MS assays. Discrimination between diets was observed by application of two linear mixed models: one including a diet-time interaction effect and the second containing only a time effect. Valerolactones, phenolic acids and their derivatives were among sixteen biomarkers related to the high flavonoid dietary exposure. Four biomarkers related to the low flavonoid diet belonged to the family of phenolic acids. For the first time abscisic acid glucuronide was reported as a biomarker after a dietary intake, however its origins have to be examined by future hypothesis driven experiments using a more targeted approach. This metabolomic analysis has identified a number of dose dependent urinary biomarkers (i.e. proline betaine or iberin-N-acetyl cysteine), which can be used in future observation and intervention studies to assess flavonoids and non-flavonoid phenolic intakes and compliance to fruit and vegetable intervention.

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Acknowledgments

FLAVURS study group: Julie A Lovegrove (PI); Anne-Marie Minihane; Michael H Gordon, Jeremy PE Spencer, Kieran M. Tuohy, Orla B Kennedy, Trevor W George (Study Co-ordinator), Mary Foong-Fong Chong (Study Co-ordinator), Dauren Alimbetov, Yin Jin and Anna Macready. The authors would like to express their sincere thanks to all those involved in the FLAVURS project and in particular, to Orla B. Kennedy, Jeremy P.E. Spencer and Mike Gordon, for their critical reading of this article.

This metabolomic study was financially supported by the ADP2014 project, funded by the Autonomous Province of Trento, Italy. The work by Jan Stanstrup was supported by a research grant (VKR023371) from VILLUM FONDEN.

Author's contribution

MU conceived and conducted the metabolomics analyses with KT, performed metabolite annotation, and wrote the body of the paper with FM, JS, KMT and JAL; JS conceived and conducted the LC–MS data analysis with PF and MU. KT helped with the data analysis; JAL conceived, obtained funding and conducted the intervention study, of which she was coordinator, with KMT, AMM, TG, MCFF and the FLAVURS study group. FM designed and coordinated the metabolomic research and obtained funding, together with KMT. All authors reviewed, edited and approved the final version of the manuscript.

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Author notes

  1. Mary Foong-Fong Chong

    Present address: Singapore Institute for Clinical Sciences, Brenner Centre, 30 Medical Drive, 117609, Singapore, Singapore

  2. Trevor George

    Present address: Department of Applied Sciences, Faculty of Health and Life Sciences, Northumbria University, Ellison Place, Newcastle upon Tyne, NE1 8ST, UK

Authors and Affiliations

  1. Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach (FEM), Via Mach 1, 38010, San Michele all’Adige, TN, Italy

    Maria M. Ulaszewska, Kajetan Trost, Jan Stanstrup, Kieran M. Tuohy, Pietro Franceschi & Fulvio Mattivi

  2. Wine Research Centre, University of Nova Gorica, Vipavska 13, 5000, Nova Gorica, Slovenia

    Kajetan Trost

  3. Hugh Sinclair Unit of Human Nutrition and the Institute for Cardiovascular and Metabolic Research (ICMR), Department of Food and Nutritional Sciences, University of Reading, Whiteknights, PO Box 226, Reading, RG6 6AP, UK

    Mary Foong-Fong Chong, Trevor George, Anne Marie Minihane & Julie A. Lovegrove

  4. Department of Nutrition, Norwich Medical School, University of East Anglia, James Watson Road, NR4 7UQ, Norwich, UK

    Anne Marie Minihane

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  1. Maria M. Ulaszewska
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  10. Fulvio Mattivi
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Correspondence to Maria M. Ulaszewska, Kieran M. Tuohy or Fulvio Mattivi.

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Ethical approval

FLAVURS was registered as a randomized clinical trial (ISRCTN47748735) and conducted according to the guidelines laid down in the Declaration of Helsinki. Ethical approval for the study was obtained from the Local Research Ethics Committee of the Isle of Wight, Portsmouth and South East Hampshire (REC: 07/H0501/81), and the University of Reading’s Research (REC: 07/22) Ethics Committee. Informed consent was obtained before participation.

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For the FLAVURS study group listed below Acknowledgment section.

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Ulaszewska, M.M., Trost, K., Stanstrup, J. et al. Urinary metabolomic profiling to identify biomarkers of a flavonoid-rich and flavonoid-poor fruits and vegetables diet in adults: the FLAVURS trial. Metabolomics 12, 32 (2016). https://doi.org/10.1007/s11306-015-0935-z

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