Evaluation of the availability of delphinidin and cyanidin-3-O-sambubioside from Hibiscus sabdariffa and 6-gingerol from Zingiber officinale in colon using liquid chromatography and mass spectrometry detection

  • Yassine Oulad El Majdoub
  • Mohammed Diouri
  • Paola Arena
  • Adriana Arigò
  • Francesco CacciolaEmail author
  • Francesca Rigano
  • Paola Dugo
  • Luigi Mondello
Original Paper


The in vivo biological activity of Hibiscus sabdariffa (H.s.) and Zingiber officinale (Z.o.) polyphenols is tightly related to their availability in the site of action. The availability of non-absorbed and intact polyphenols, or partially metabolized, in the colon, would perform, according to in vitro outcomes, a highly antioxidant activity and trapping effect towards toxic molecules (e.g., methylglyoxal), which are locally produced in the colon by the gut microbiota. The aim of this study was to evaluate the in vivo availability of three selected polyphenolic compounds of dried H.s calyces, and fresh Z.o. rhizomes in the Wistar rat colon by liquid chromatography coupled to photodiode array and mass spectrometry detection (HPLC–PDA/ESI–MS). The colon-available polyphenols were extracted through the initial solid–liquid faeces extraction by the employment of a solvent mixture (methanol:water, 60:40 v/v), followed by a solid-phase extraction (SPE) on a C18 cartridge. The main H.s. anthocyanins (cyanidin-3-O-sambubioside, C3S and delphinidin-3-O-sambubioside, D3S) and 6-gingerol of Z.o. were available in the intact form in the colon 12 h after the administration of concentrated aqueous extracts (6% and 4% w/v). 72.15% of the ingested C3S and 76.19% of D3S were available in the colon, in comparison to the low availability of 6-gingerol equal to 1.50%. The duration of these bioactive compounds availability in the colon was limited to 12 h. The anthocyanin and gingerol availability in the colon may favor their absorption into the enterocytes, contributing to the antioxidant potential and health effects.


Anthocyanins Gingerol Zingiber officinale Hibiscus sabdariffa Availability Colon 



The authors are thankful to Shimadzu and Merck Life Science Corporations for the continuous support.

Compliance with ethical standards

Conflict of interest

The authors of this work have declared no conflict of interest.

Ethical standards

All animal procedures were in accordance with the Guidelines of Italian Health Minister (D.M: 116192), as well as with the EEC regulations (O.J. of E.C.L. 358/1 12/18/1986).

Supplementary material

217_2019_3358_MOESM1_ESM.docx (727 kb)
Supplementary material 1 (DOCX 726 kb)


  1. 1.
    Campbell AK, Matthews SB, Vassel N et al (2010) Bacterial metabolic ‘toxins’: a new mechanism for lactose and food intolerance, and irritable bowel syndrome. Toxicology 278:268–276CrossRefGoogle Scholar
  2. 2.
    Campbell AK, Waud JP, Matthews SB (2005) The molecular basis of lactose intolerance. Sci Prog 88:157–202CrossRefGoogle Scholar
  3. 3.
    Campbell AK, Naseem R, Holland IB et al (2007) Methylglyoxal and other carbohydrate metabolites induce lanthanum-sensitive Ca2+ transients and inhibit growth in E. coli. Arch Biochem Biophys 468:107–113CrossRefGoogle Scholar
  4. 4.
    Ho C-T, Wang M (2013) Dietary phenolics as reactive carbonyl scavengers: potential impact on human health and mechanism of action. J Trad Complement Med 3:139–141CrossRefGoogle Scholar
  5. 5.
    Zhu Y, Zhao Y, Wang P et al (2015) Bioactive ginger constituents alleviate protein glycation by trapping methylglyoxal. Chem Res Toxicol 28:1842–1849CrossRefGoogle Scholar
  6. 6.
    Kumar S, Pandey AK (2007) Chemistry and biological activities of flavonoids: an overview. Sci World J 73:637–670Google Scholar
  7. 7.
    Tsao R (2010) Chemistry and biochemistry of dietary polyphenols. Nutrients 2:1231–1246CrossRefGoogle Scholar
  8. 8.
    Crozier A, Jaganath IB, Clifford MN (2009) Dietary phenolics: chemistry, bioavailability and effects on health. Nat Prod Rep 26:1001–1043CrossRefGoogle Scholar
  9. 9.
    Morohashi K, Casas MI, Ferreyra MLF, Mejia-Guerra MK, Pourcel L, Yilmaz A, Feller A, Carvalho B, Emiliani J, Rodriguez E et al (2012) A genome-wide regulatory framework identifies maize pericarp Color1 controlled genes. Plant Cell 24:2745–2764CrossRefGoogle Scholar
  10. 10.
    Fantini M, Benvenuto M, Masuelli L, Frajese G, Tresoldi I, Modesti A, Bei R (2015) In vitro and in vivo antitumoral effects of combinations of polyphenols, or polyphenols and anticancer drugs: perspectives on cancer treatment. Int J Mol Sci 16:9236–9282CrossRefGoogle Scholar
  11. 11.
    Grajeda-Iglesias C, Figueroa-Espinoza MC, Barouh N et al (2016) Isolation and characterization of anthocyanins from Hibiscus sabdariffa flowers. J Nat Prod 79:1709–1718CrossRefGoogle Scholar
  12. 12.
    Baliga MS, Latheef L, Haniadka R, Fazal F, Chacko J, Arora R (2013) Chapter 41-ginger (Zingiber officinale Roscoe) in the treatment and prevention of arthritis. In: Preedy RRWR (ed) Bioactive food as dietary interventions for arthritis and related inflammatory diseases. Academic Press, San Diego, pp 529–544CrossRefGoogle Scholar
  13. 13.
    Jolad SD, Lantz RC, Solyom AM et al (2004) Fresh organically grown ginger (Zingiber officinale): composition and effects on LPS-induced PGE2 production. Phytochemistry 65:1937–1954CrossRefGoogle Scholar
  14. 14.
    Ichiyanagi T, Shida Y, Rahman MM, Hatano Y, Konishi T (2005) Extended glucuronidation is another major path of cyanidin 3-O-b-D-glucopyranoside metabolism in rats. J Agric Food Chem 53:7312–7319CrossRefGoogle Scholar
  15. 15.
    Cardona F, Andrés-Lacueva C, Tulipani S et al (2013) Benefits of polyphenols on gut microbiota and implications in human health. J Nutr Biochem 24:1415–1422CrossRefGoogle Scholar
  16. 16.
    Sogo T, Terahara N, Hisanaga A et al (2015) Anti-inflammatory activity and molecular mechanism of delphinidin 3-sambubioside, a Hibiscus anthocyanin: anti-inflammatory effects of delphinidin 3-sambubioside. BioFactors 41:58–65CrossRefGoogle Scholar
  17. 17.
    Ghasemzadeh A, Jaafar HZE, Rahmat A (2016) Changes in antioxidant and antibacterial activities as well as phytochemical constituents associated with ginger storage and polyphenol oxidase activity. BMC Complement Altern Med 16:382CrossRefGoogle Scholar
  18. 18.
    Official Methods of Analysis of Analysis Association of Official Analytical Chemists International AOAC (1996). Moisture in animal feed, method 930.15 (16th edn), Gaithersburg, MD. References—Scientific Research Publishing. n.d. Accessed 4 Dec 2018Google Scholar
  19. 19.
    He J, Magnuson BA, Giusti MM (2005) Analysis of anthocyanins in rat intestinal contents impact of anthocyanin chemical structure on fecal excretion. J Agric Food Chem 53:2859–2866CrossRefGoogle Scholar
  20. 20.
    Zick SM, Ruffin MT, Djuric Z et al (2010) Quantitation of 6-, 8- and 10-gingerols and 6-shogaol in human plasma by high-performance liquid chromatography with electrochemical detection. Int J Biomed Sci 6:8Google Scholar
  21. 21.
    National Sanitation Foundation (NSF) HPLC Analysis of Gingerols and Shogaols in Zingiber officinale (Ginger) INA Method 114.000 International. Accessed 9 Jul 2010
  22. 22.
    Oyedemi BOM, Maria KE, Stapleton PD, Gibbons S (2019) Capsaicin and gingerol analogues inhibit the growth of efflux-multidrug resistant bacteria and R-plasmids conjugal transfer. J Ethnopharmacol 2019:111871CrossRefGoogle Scholar
  23. 23.
    Kallam K, Appelhagen I, Luo J et al (2017) Aromatic decoration determines the formation of anthocyanic vacuolar inclusions. Curr Biol 27:945–957CrossRefGoogle Scholar
  24. 24.
    Fanali C, Dugo L, D’Orazio G et al (2011) Analysis of anthocyanins in commercial fruit juices by using nano-liquid chromatography-electrospray-mass spectrometry and high-performance liquid chromatography with UV–vis detector. J Sep Sci 34:150–159CrossRefGoogle Scholar
  25. 25.
    Cesa S, Carradori S, Bellagamba G et al (2017) Evaluation of processing effects on anthocyanin content and colour modifications of blueberry (Vaccinium spp.) extracts: comparison between HPLC-DAD and CIELAB analyses. Food Chem 232:114–123CrossRefGoogle Scholar
  26. 26.
    Aura A-M, Martin-Lopez P, O’Leary KA et al (2005) In vitro metabolism of anthocyanins by human gut microflora. Eur J Nutr 44:133–142CrossRefGoogle Scholar
  27. 27.
    Matuschek MC, Hendriks WH, McGhie TK, Reynolds GW (2006) The jejunum is the main site of absorption for anthocyanins in mice. J Nutr Biochem 17(1):31–36CrossRefGoogle Scholar
  28. 28.
    Williamson G, Kay CD, Crozier A (2018) The bioavailability, transport, and bioactivity of dietary flavonoids: a review from a historical perspective: dietary flavonoids: a historical review. Compr Rev Food Sci F 17(5):1054–1112CrossRefGoogle Scholar
  29. 29.
    Wu X, Pittman HE, Prior RL (2006) Fate of anthocyanins and antioxidant capacity in contents of the gastrointestinal tract of weanling pigs following black raspberry consumption. J Agric Food Chem 54:583–589CrossRefGoogle Scholar
  30. 30.
    Esposito D, Damsud T, Wilson M, Grace MH, Strauch R, Li X, Lila MA, Komarnytsky S (2015) Black currant anthocyanins attenuate weight gain and improve glucose metabolism in diet-induced obese mice with intact, but not disrupted, gut microbiome. J Agric Food Chem 63(27):6172–6180CrossRefGoogle Scholar
  31. 31.
    McGhie TK, Ainge GD, Barnett LE, Cooney JM, Jensen JD (2003) Anthocyanin glycosides from berry fruit are absorbed and excreted unmetabolized by both humans and rats. J Agric Food Chem 51:4539–4548CrossRefGoogle Scholar
  32. 32.
    Miyazawa T, Nakagawa K, Kudo M, Muraishi K, Someya K (1999) Direct intestinal absorption of red fruit anthocyanins, cyanidin-3-glucoside and cyanidin-3,5-diglucoside, into rats and humans. J Agric Food Chem 47:1083–1091CrossRefGoogle Scholar
  33. 33.
    Milbury P, Cao G, Prior RL, Blumberg J (2002) Bioavailability of elderberry anthocyanins. Mech Ageing Dev 123:997–1006CrossRefGoogle Scholar
  34. 34.
    Cooney JM, Jensen JD, McGhie TK (2004) LC–MS identification of anthocyanins in boysenberry extract and anthocyanin metabolites in human urine following dosing. J Sci Food Agric 84:237–245CrossRefGoogle Scholar
  35. 35.
    Wu X, Pittman HE, Mckay S, Prior RL (2005) Aglycones and sugar moieties alter anthocyanin absorption and metabolism after berry consumption in weanling pigs. J Nutr 135(10):2417–2424CrossRefGoogle Scholar
  36. 36.
    Gonzalez-Barrio R, Edwards CA, Crozier A (2011) Colonic catabolism of ellagitannins, ellagic acid, and raspberry anthocyanins. In vivo and in vitro studies. Drug Metab Dispos 39(9):1680–1688CrossRefGoogle Scholar
  37. 37.
    McDougall GJ, Conner S, Pereira-Caro G, Gonzalez-Barrio R, Brown EM, Verrall S, Stewart D et al (2014) Tracking (poly)phenol components from raspberries in ileal fluid. J Agric Food Chem 62(30):7631–7641CrossRefGoogle Scholar
  38. 38.
    Feliciano RP, Boeres A, Massacessi L, Istas G, Ventura MR, dos Santos CN, Heiss C, Rodriguez-Mateos A (2016) Identification and quantification of novel cranberry-derived plasma and urinary (poly)phenols. Arch Biochem Biophys 599(June):31–41CrossRefGoogle Scholar
  39. 39.
    Rodriguez-Mateos A, Vauzour D, Krueger CG, Shanmuganayagam D, Reed J, Calani L, Mena P, Del Rio D, Crozier A (2014) Bioavailability, bioactivity and impact on health of dietary flavonoids and related compounds: an update. Arch Toxicol 88(10):1803–1853CrossRefGoogle Scholar
  40. 40.
    Nakazawa T, Ohsawa K (2002) Metabolism of [6]-gingerol in rats. Life Sci 70(18):2165–2175CrossRefGoogle Scholar
  41. 41.
    Ahmed RS, Seth V, Banerjee BD (2000) Influence of dietary ginger (Zingiber officinale Rose) on antioxidant defense-system in rat: comparison with ascorbic acid. Food Chem Toxicol 3:443CrossRefGoogle Scholar
  42. 42.
    Zick SM, Djuric Z, Ruffin MT et al (2008) Pharmacokinetics of 6-gingerol, 8-gingerol, 10-gingerol, and 6-shogaol and conjugate metabolites in healthy human subjects. Cancer Epidemiol Biomark Prev 17:1930–1936CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Yassine Oulad El Majdoub
    • 1
  • Mohammed Diouri
    • 1
  • Paola Arena
    • 2
  • Adriana Arigò
    • 2
  • Francesco Cacciola
    • 3
    Email author
  • Francesca Rigano
    • 4
  • Paola Dugo
    • 2
    • 4
    • 5
  • Luigi Mondello
    • 2
    • 4
    • 5
    • 6
  1. 1.Laboratory of Plant Biotechnologies and Molecular Biology, Department of Biology, Faculty of Sciences of MeknesMoulay Ismail UniversityMeknesMorocco
  2. 2.Department of Chemical, Biological, Pharmaceutical and Environmental SciencesUniversity of MessinaMessinaItaly
  3. 3.Department of Biomedical, Dental, Morphological and Functional Imaging SciencesUniversity of MessinaMessinaItaly
  4. 4.Chromaleont s.r.l., c/o Department of Chemical, Biological, Pharmaceutical and Environmental SciencesUniversity of MessinaMessinaItaly
  5. 5.Unit of Food Science and Nutrition, Department of MedicineUniversity Campus Bio-Medico of RomeRomeItaly
  6. 6.BeSep s.r.l., c/o Department of Chemical, Biological, Pharmaceutical and Environmental SciencesUniversity of MessinaMessinaItaly

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