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Nuclear magnetic resonance and high-performance liquid chromatography techniques for the characterization of bioactive compounds from Humulus lupulus L. (hop)

Analytical and Bioanalytical Chemistry volume 410, pages 3521–3531 (2018)Cite this article

Abstract

Humulus lupulus L. (hop) represents one of the most cultivated crops, it being a key ingredient in the brewing process. Many health-related properties have been described for hop extracts, making this plant gain more interest in the field of pharmaceutical and nutraceutical research. Among the analytical tools available for the phytochemical characterization of plant extracts, quantitative nuclear magnetic resonance (qNMR) represents a new and powerful technique. In this ambit, the present study was aimed at the development of a new, simple, and efficient qNMR method for the metabolite fingerprinting of bioactive compounds in hop cones, taking advantage of the novel ERETIC 2 tool. To the best of our knowledge, this is the first attempt to apply this method to complex matrices of natural origin, such as hop extracts. The qNMR method set up in this study was applied to the quantification of both prenylflavonoids and bitter acids in eight hop cultivars. The performance of this analytical method was compared with that of HPLC-UV/DAD, which represents the most frequently used technique in the field of natural product analysis. The quantitative data obtained for hop samples by means of the two aforementioned techniques highlighted that the amount of bioactive compounds was slightly higher when qNMR was applied, although the order of magnitude of the values was the same. The accuracy of qNMR was comparable to that of the chromatographic method, thus proving to be a reliable tool for the analysis of these secondary metabolites in hop extracts.

Graphical abstract related to the extraction and analytical methods applied in this work for the analysis of bioactive compounds in Humulus lupulus L. (hop) cones

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References

  1. Alekseeva MA, Éller KI, Arzamastsev AP. Determining polyphenolic components of common hop by reversed-phase HPLC. Pharm Chem J. 2004;38:687–9.

    Article  CAS  Google Scholar 

  2. Van Cleemput M, Cattoor K, De Bosscher K, Haegeman G, De Keukeleire D, Heyerick A. Hop (Humulus lupulus)-derived bitter acids as multipotent bioactive compounds. J Nat Prod. 2009;72:1220–30.

    Article  CAS  PubMed  Google Scholar 

  3. Magalhaes PJ, Vieira JS, Goncalves LM, Pacheco JG, Guido LF, Barros AA. Isolation of phenolic compounds from hop extracts using polyvinylpolypyrrolidone: characterization by high-performance liquid chromatography-diode array detection-electrospray tandem mass spectrometry. J Chromatogr A. 2010;1217:3258–68.

    Article  CAS  PubMed  Google Scholar 

  4. Karabín M, Huducová T, Jelínek L, Dostálek P. Biologically active compounds from hops and prospects for their use. Compr Rev Food Sci Food Saf. 2016;15:542–67.

    Article  CAS  Google Scholar 

  5. Kavalier AR, Litt A, Ma C, Pitra NJ, Coles MC, Kennelly EJ, et al. Phytochemical and morphological characterization of hop (Humulus lupulus L.) cones over five developmental stages using high performance liquid chromatography coupled to time-of-flight mass spectrometry, ultrahigh performance liquid chromatography photodiode array detection, and light microscopy techniques. J Agric Food Chem. 2011;59:4783–93.

  6. Taniguchi Y, Taniguchi H, Yamada M, Matsukura Y, Koizumi H, Furihata K, et al. Analysis of the component of the hard resin in hops (Humulus lupulus L.) and structural elucidation of their transformation products formed during the brewing process. J Agric Food Chem. 2014;62:11602–12.

  7. Ceslová L, Holcapek M, Fidler M, Drsticková J, Lísa M. Characterization of prenylflavonoids and hop bitter acids in various classes of Czech beers and hop extracts using high-performance liquid chromatography-mass spectrometry. J Chromatogr A. 2009;1216:7249–57.

    Article  CAS  PubMed  Google Scholar 

  8. Martino F, Pace S, Chianese G, Santagostini L, Werner M, Weinigel C, et al. Humudifucol and bioactive prenylated polyphenols from hops (Humulus lupulus cv. “Cascade”). J Nat Prod. 2016;79:590–7.

  9. Almaguer C, Schönberger C, Gastl M, Arendt EK, Becker T. Humulus lupulus—a story that begs to be told. A review. J Inst Brew. 2014;120:289–314.

    CAS  Google Scholar 

  10. Brunelli E, Minassi A, Appendino G, Moro L. 8-Prenylnaringenin, inhibits estrogen receptors-α mediated cell growth and induces apoptosis in MCF-7 breast cancer cells. J Steroid Biochem Mol Biol. 2007;107:140–8.

    Article  CAS  PubMed  Google Scholar 

  11. Keiler AM, Zierau O, Kretzschmar G. Hop extracts and hop substances in treatment of menopausal complaints. Planta Med. 2013;79:576–9.

    Article  CAS  PubMed  Google Scholar 

  12. Jelínek L, Šneberger M, Karabín M, Dostálek P. Comparison of Czech hop cultivars based on their contents of secondary metabolites. Czech J Food Sci. 2010;28:309–16.

    Article  Google Scholar 

  13. Dhooge L, Naessens T, Heyerick A, De Keukeleire D, Vlietinck AJ, Pieters L, et al. Quantification of xanthohumol, isoxanthohumol, 8-prenylnaringenin and 6-prenylnaringenin in hop extracts and derived capsules using secondary standards. Talanta. 2010;83:448–56.

  14. Prencipe FP, Brighenti V, Rodolfi M, Mongelli A, Dall’Asta C, Ganino T, et al. Development of a new high-performance liquid chromatography method with diode array and electrospray ionization-mass spectrometry detection for the metabolite fingerprinting of bioactive compounds in Humulus lupulus L. J Chromatogr A. 2014;1349:50–9.

  15. Nikolic D, Van Breemen R. Analytical methods for quantitation of prenylated flavonoids from hops. Curr Anal Chem. 2013;9:71–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Marcone MF, Wang S, Albabish W, Nie S, Somnarain D, Hill A. Diverse food-based applications of nuclear magnetic resonance. Food Res Int. 2013;51:729–47.

    Article  CAS  Google Scholar 

  17. Höltzel A, Schlotterbeck G, Albert K, Bayer E. Separation and characterization of hop bitter acids by HPLC-1HNMR coupling. Chromatographia. 1996;42:500–5.

    Article  Google Scholar 

  18. De Keukeleire J, Ooms G, Heyerick A, Roldan-Ruiz I, Van Bockstaele E, de Keukeleire D. Formation and accumulation of α-acids, β-acids, desmethylxanthohumol and xanthohumol during flowering of hops (Humulus lupulus L.). J Agric Food Chem. 2003;51:4436–41.

    Article  CAS  PubMed  Google Scholar 

  19. Taniguchi Y, Matsukura Y, Ozaki H, Nishimura K, Shindo K. Identification and quantification of the oxidation products derived from α-acids and β-acids during storage of hops (Humulus lupulus L.). J Agric Food Chem. 2013;61:3121–30.

    Article  CAS  PubMed  Google Scholar 

  20. Farag MA, Porzel A, Scmidt J, Wessjohan LA. Metabolite profiling and fingerprinting of commercial cultivars of Humulus lupulus L. (hop): a comparison of MS and NMR methods in metabolomics. Metabolomics. 2012;8:492–507.

    Article  CAS  Google Scholar 

  21. Farag MS, Mahrous EA, Lübken T, Porzel A, Wessjohan LA. Classification of commercial cultivars of Humulus lupulus L. (hop) by chemometric pixel analysis of two dimensional nuclear magnetic resonance spectra. Metabolomics. 2013;10:21–32.

    Article  CAS  Google Scholar 

  22. Kileen DP, Watkins OC, Sansom CE, Andersen DH, Gordon KC, Perry NB. Fast sampling, analyses and chemometrics for plant breeding: bitter acids, xanthohumol and terpenes in lupulin glands of hops (Humulus lupulus). Phytochem Anal. 2017;28:50–7.

    Article  CAS  Google Scholar 

  23. Bharti SK, Roy R. Quantitative 1H NMR spectroscopy. Trend Anal Chem. 2012;35:5–26.

    Article  CAS  Google Scholar 

  24. Zołnierczyk AK, Mączka WK, Grabarczyk M, Wińska K, Woźniak E, Anioł M. Isoxanthohumol—biologically active hop flavonoid. Fitoterapia. 2015;103:71–82.

    Article  CAS  PubMed  Google Scholar 

  25. Molyneux RJ, Wong Y-I. Nuclear magnetic resonance spectroscopic determination of α- and β-acid homolog composition in hops. J Agric Food Chem. 1975;23:1201–4.

    Article  CAS  Google Scholar 

  26. Watanabe R, Sugai C, Yamazaki T, Matsushima R, Uchida H, Matsumiya M, et al. Quantitative nuclear magnetic resonance spectroscopy based on PULCON methodology: application to quantification of invaluable marine toxin, okadaic acid. Toxins. 2016;8:294–302.

  27. Le Grand F, George G, Akoka S. How to reduce the experimental time in isotopic 2H NMR using the ERETIC method. J Magn Reson. 2005;174:171–6.

    Article  CAS  PubMed  Google Scholar 

  28. Botta B, Vitali A, Menendez P, Misiti D, Monache GD. Prenylated flavonoids: pharmacology and biotechnology. Curr Med Chem. 2005;12:713–39.

    Article  CAS  Google Scholar 

  29. Gil-Ramírez A, Mendiola JA, Arranz E, Ruíz-Rodríguez A, Reglero G, Ibáñez E, et al. Highly xanthohumol enriched hop extract obtained by pressurized hot water extraction (PHWE). Chemical and functional characterization. Innovative Food Sci Emerging Technol. 2012;16:54–60.

  30. Magalhães PJ, Guido LF, Cruz JM, Barros AA. Analysis of xanthohumol and isoxanthohumol in different hop products by liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry. J Chromatogr A. 2007;1150:295–301.

    Article  CAS  PubMed  Google Scholar 

  31. Dresel M, Vogt C, Dunkel A, Hofmann T. The bitter chemodiversity of hops (Humulus lupulus L.). J Agric Food Chem. 2016;64:7789–99.

    Article  CAS  Google Scholar 

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Acknowledgments

This work was carried out thanks to a grant awarded to Anna Reik for her master thesis work in the lab of Dr. Federica Pellati and Dr. Davide Bertelli at the Department of Life Sciences of the University of Modena and Reggio Emilia (Italy), in the ambit of the Erasmus+ Program.

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Affiliations

  1. Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, 41125, Modena, Italy

    Davide Bertelli, Virginia Brighenti, Lucia Marchetti, Anna Reik & Federica Pellati

  2. Faculty of Agricultural Sciences, Nutritional Sciences, and Environmental Management, Justus-Liebig University of Giessen, Goethestrasse 58, 35390, Giessen, Germany

    Anna Reik

Authors
  1. Davide Bertelli
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  2. Virginia Brighenti
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  3. Lucia Marchetti
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  4. Anna Reik
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  5. Federica Pellati
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Correspondence to Federica Pellati.

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Published in the topical collection Discovery of Bioactive Compounds with guest editors Aldo Laganà, Anna Laura Capriotti and Chiara Cavaliere.

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Cite this article

Bertelli, D., Brighenti, V., Marchetti, L. et al. Nuclear magnetic resonance and high-performance liquid chromatography techniques for the characterization of bioactive compounds from Humulus lupulus L. (hop). Anal Bioanal Chem 410, 3521–3531 (2018). https://doi.org/10.1007/s00216-018-0851-y

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  • DOI: https://doi.org/10.1007/s00216-018-0851-y

Keywords

  • Humulus lupulus L.
  • Hop
  • Prenylflavonoids
  • Bitter acids
  • qNMR
  • HPLC