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Discovery of exposure markers in urine for Brassica-containing meals served with different protein sources by UPLC-qTOF-MS untargeted metabolomics

Metabolomics volume 9, pages 984–997 (2013)Cite this article

Abstract

An untargeted metabolomics approach has been applied to discover and identify exposure markers in urine for nine Nordic meals. A cross-over meal study was carried out in 17 subjects. The meals included a Pie, a Soup and a Barleyotto (pearl barley based risotto), each prepared with three protein sources; meat, fish or vegetarian. Urine samples were collected in different time intervals before and after intake of the test meals, covering a total of 24 h. The samples were analyzed by UPLC-qTOF-MS. Discriminating features for meals and protein sources were selected by use of double cross-validated partial least squares discriminant analysis and two additional validation steps: (1) time-course of excretion and (2) analysis of sensitivity and specificity. In addition, eight meal studies with single foods were carried out to investigate the food sources of the markers. In total 31 potential exposure markers (PEMs) of foods were found for the meals and protein sources. Fifteen of the 31 PEMs were also found in studies with single foods. Ten PEMs were identified or putatively annotated. Among the PEMs were a range of conjugated isothiocyanates from the Brassica oleracea species. Trimethylamine N-oxide was found as a fish marker. Additional unknown PEMs were found for chicory salad, parsley and fava beans, while other PEMs were dependent on the meal matrix rather than individual foods. The study demonstrates that it is possible to find PEMs in 24 h urine samples even when foods are given as part of a complex meal.

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References

  • Agudo, A., Bailey, G. S., Bradlow, H. L., et al. (2004). Cruciferous vegetables, isothiocyanates and indoles. Lyon: International Agency for Research, on Cancer. IARC Press.

    Google Scholar 

  • Barri, T., Holmer-Jensen, J., Hermansen, K., & Dragsted, L. O. (2012). Metabolic fingerprinting of high-fat plasma samples processed by centrifugation- and filtration-based protein precipitation delineates significant differences in metabolite information coverage. Analytica Chimica Acta, 718, 47–57.

    PubMed  Article  CAS  Google Scholar 

  • Bijlsma, S., Bobeldijk, I., Verheij, E. R., et al. (2006). Large-scale human metabolomics studies: A strategy for data (pre-) processing and validation. Analytical Chemistry, 78, 567–574.

    PubMed  Article  CAS  Google Scholar 

  • Bingham, S. A. (2002). Biomarkers in nutritional epidemiology. Public Health Nutrition, 5, 821–827.

    PubMed  Article  Google Scholar 

  • Chong, I., & Jun, C. (2005). Performance of some variable selection methods when multicollinearity is present. Chemometrics and Intelligent Laboratory Systems, 78, 103–112.

    Article  CAS  Google Scholar 

  • Clarke, D. B. (2010). Glucosinolates, structures and analysis in food. Analytical Methods, 2, 310–325.

    Article  CAS  Google Scholar 

  • Favé, G., Beckmann, M. E., Draper, J. H., & Mathers, J. C. (2009). Measurement of dietary exposure: A challenging problem which may be overcome thanks to metabolomics? Genes and Nutrition, 4, 135–141.

    PubMed  Article  Google Scholar 

  • Favé, G., Beckmann, M., Lloyd, A., et al. (2011). Development and validation of a standardized protocol to monitor human dietary exposure by metabolite fingerprinting of urine samples. Metabolomics, 7, 469–484.

    PubMed  Article  Google Scholar 

  • Holmes, E., Loo, R. L., Stamler, J., et al. (2008). Human metabolic phenotype diversity and its association with diet and blood pressure. Nature, 453, 396–400.

    PubMed  Article  CAS  Google Scholar 

  • Hwang, E. S., & Jeffery, E. H. (2003). Evaluation of urinary N-acetyl cysteinyl allyl isothiocyanate as a biomarker for intake and bioactivity of Brussels sprouts. Food and Chemical Toxicology, 41, 1817–1825.

    PubMed  Article  CAS  Google Scholar 

  • Janobi, A. A. A., Mithen, R. F., Gasper, A. V., et al. (2006). Quantitative measurement of sulforaphane, iberin and their mercapturic acid pathway metabolites in human plasma and urine using liquid chromatography–tandem electrospray ionisation mass spectrometry. Journal of Chromatography B, 844, 223–234.

    Article  Google Scholar 

  • Jenab, M., Slimani, N., Bictash, M., et al. (2009). Biomarkers in nutritional epidemiology: Applications, needs and new horizons. Human Genetics, 125, 507–525.

    PubMed  Article  Google Scholar 

  • Koulman, A., & Volmer, D. A. (2008). Perspectives for metabolomics in human nutrition: An overview. Nutrition Bulletin, 33, 324–330.

    PubMed  Article  Google Scholar 

  • Liu, Z., & Franklin, M. R. (1984). Separation of four glucuronides in a single sample by high-pressure liquid chromatography and its use in the determination of UDP glucuronosyltransferase activity toward four aglycones. Analytical Biochemistry, 142, 340–346.

    PubMed  Article  CAS  Google Scholar 

  • Llorach, R., Garrido, I., Monagas, M., et al. (2010). Metabolomics study of human urinary metabolome modifications after intake of almond (Prunus dulcis (Mill.) DA Webb) skin polyphenols. Journal of Proteome Research, 9, 5859–5867.

    PubMed  Article  CAS  Google Scholar 

  • Lloyd, A. J., Fave, G., Beckmann, M., et al. (2011). Use of mass spectrometry fingerprinting to identify urinary metabolites after consumption of specific foods. American Journal of Clinical Nutrition, 94, 981–991.

    PubMed  Article  CAS  Google Scholar 

  • Lodge, J. K. (2010). Symposium 2: Modern approaches to nutritional research challenges: Targeted and non-targeted approaches for metabolite profiling in nutritional research. Proceedings of the Nutrition Society, 69, 95–102.

    PubMed  Article  CAS  Google Scholar 

  • Mithril, C., Dragsted, L. O., Meyer, C., Blauert, E., Holt, M. K., & Astrup, A. (2012a). Guidelines for the new Nordic diet. Public Health Nutrition, 15, 1941–1947.

    PubMed  Article  Google Scholar 

  • Mithril, C, Dragsted, L. O., Meyer, C., Tetens, I., Jensen, A. B. & Astrup, A. (2012b). Dietary composition and nutrient content of the new Nordic diet, in press.

  • Nagata, C., Takatsuka, N., & Shimizu, H. (2002). Soy and fish oil intake and mortality in a Japanese community. American Journal of Epidemiology, 156, 824–831.

    PubMed  Article  Google Scholar 

  • Nelson, A. C., Huang, W., & Moody, D. E. (2001). Variables in human liver microsome preparation: Impact on the kinetics of l-alpha-acetylmethadol (LAAM) n-demethylation and dextromethorphan O-demethylation. Drug Metabolism and Disposition, 29, 319–325.

    PubMed  CAS  Google Scholar 

  • O’Sullivan, A., Gibney, M. J., & Brennan, L. (2011). Dietary intake patterns are reflected in metabolomic profiles: Potential role in dietary assessment studies. American Journal of Clinical Nutrition, 93, 314–321.

    PubMed  Article  Google Scholar 

  • Penn, L., Boeing, H., Boushey, C., et al. (2010). Assessment of dietary intake: NuGO symposium report. Genes and Nutrition, 5, 205–213.

    PubMed  Article  CAS  Google Scholar 

  • Pluskal, T., Castillo, S., Villar-Briones, A., & Orešič, M. (2010). MZmine 2: Modular framework for processing, visualizing, and analyzing mass spectrometry-based molecular profile data. BMC Bioinformatics, 11, 395.

    PubMed  Article  Google Scholar 

  • Puiggròs, F., Solà, R., Bladé, C., Salvadó, M., & Arola, L. (2011). Nutritional biomarkers and foodomic methodologies for qualitative and quantitative analysis of bioactive ingredients in dietary intervention studies. Journal of Chromatography A, 1218, 7399–7414.

    PubMed  Article  Google Scholar 

  • Rouzaud, G., Young, S. A., & Duncan, A. J. (2004). Hydrolysis of glucosinolates to isothiocyanates after ingestion of raw or microwaved cabbage by human volunteers. Cancer Epidemiology, Biomarkers and Prevention, 13, 125–131.

    PubMed  Article  CAS  Google Scholar 

  • Solanky, K. S., Bailey, N. J., Beckwith-Hall, B. M., et al. (2005). Biofluid 1H NMR-based metabonomic techniques in nutrition research: Metabolic effects of dietary isoflavones in humans. Journal of Nutritional Biochemistry, 16, 236–244.

    PubMed  Article  CAS  Google Scholar 

  • Spencer, J. P. E., Abd El Mohsen, M. M., Minihane, A., et al. (2008). Biomarkers of the intake of dietary polyphenols: Strengths, limitations and application in nutrition research. British Journal of Nutrition, 99(1), 12–22.

    PubMed  Article  CAS  Google Scholar 

  • Stella, C., Beckwith-hall, B., Cloarec, O., et al. (2006). Susceptibility of human metabolic phenotypes to dietary modulation. Journal of Proteome Research, 5, 2780–2788.

    PubMed  Article  CAS  Google Scholar 

  • Sumner, L., Amberg, A., Barrett, D., Beale, M. H., et al. (2007). Proposed minimum reporting standards for chemical analysis. Metabolomics, 3, 211–221.

    Article  CAS  Google Scholar 

  • Svensson, B. G., Åkesson, B., Nilsson, A., & Paulsson, K. (1994). Urinary excretion of methylamines in men with varying intake of fish from the Baltic sea. Journal of Toxicology and Environment Health, 41, 411–420.

    Article  CAS  Google Scholar 

  • Vermeulen, M., Van Den Berg, R., Freidig, A. P., Van Bladeren, P. J., & Vaes, W. H. J. (2006). Association between consumption of cruciferous vegetables and condiments and excretion in urine of isothiocyanate mercapturic acids. Journal of Agriculture and Food Chemistry, 54, 5350–5358.

    Article  CAS  Google Scholar 

  • Walsh, M. C., Brennan, L., Pujos-Guillot, E., et al. (2007). Influence of acute phytochemical intake on human urinary metabolomic profiles. American Journal of Clinical Nutrition, 86, 1687–1693.

    PubMed  CAS  Google Scholar 

  • Wold, S., Johansson, E., & Cocchi, M. (1993). PLS—partial least-squares projections to latent structures. In H. Kubinyi (Ed.), 3D QSAR in drug design, theory, methods and applications (pp. 523–550). Leiden: ESCOM Leiden.

    Google Scholar 

  • Xu, J., Yang, S., Cai, S., Dong, J., Li, X., & Chen, Z. (2010). Identification of biochemical changes in lactovegetarian urine using 1H NMR spectroscopy and pattern recognition. Analytical and Bioanalytical Chemistry, 396, 1451–1463.

    PubMed  Article  CAS  Google Scholar 

  • Zhang, Y. (2004). Cancer-preventive isothiocyanates: Measurement of human exposure and mechanism of action. Mutation Research, 555, 173–190.

    PubMed  Article  CAS  Google Scholar 

  • Zhang, A. Q., Mitchell, S. C., & Smith, R. L. (1999). Dietary precursors of trimethylamine in man: A pilot study. Food and Chemical Toxicology, 37, 515–520.

    PubMed  Article  CAS  Google Scholar 

  • Zuppi, C., Messana, I., Forni, F., Ferrari, F. B., Rossi, C. B., & Giardina, B. (1998). Influence of feeding on metabolite excretion evidenced by urine 1H NMR spectral profiles: A comparison between subjects living in Rome and subjects living at arctic latitudes (Svaldbard). Clinica Chimica Acta, 278, 75–79.

    Article  Google Scholar 

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Acknowledgments

The meal study was conducted as part of the OPUS project and is supported by a Grant from the Nordea Foundation. We would like to thank the study participants and the staff who have been involved in conducting the study, preparing the meals and analyzing the samples: Hanne Lysdal Petersen, Ümmühan Celik, Daniela Rago, Jan Stanstrup, the kitchen staff at the department and Meyers Madhus.

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Authors and Affiliations

  1. Department of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Rolighedsvej 30, 1958, Frederiksberg C, Denmark

    Maj-Britt Schmidt Andersen, Thaer Barri, Charlotte Mithril & Lars Ove Dragsted

  2. Department of Food Science, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark

    Helene Christine Reinbach & Åsmund Rinnan

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  1. Maj-Britt Schmidt Andersen
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  2. Helene Christine Reinbach
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  3. Åsmund Rinnan
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  4. Thaer Barri
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  5. Charlotte Mithril
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  6. Lars Ove Dragsted
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Correspondence to Maj-Britt Schmidt Andersen.

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Andersen, MB.S., Reinbach, H.C., Rinnan, Å. et al. Discovery of exposure markers in urine for Brassica-containing meals served with different protein sources by UPLC-qTOF-MS untargeted metabolomics. Metabolomics 9, 984–997 (2013). https://doi.org/10.1007/s11306-013-0522-0

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  • DOI: https://doi.org/10.1007/s11306-013-0522-0

Keywords

  • Food exposure markers
  • Meal study
  • Metabolomics
  • UPLC-QTOF-MS
  • Urine
  • Multivariate analysis