European Journal of Nutrition

, Volume 43, Issue 4, pp 205–220 | Cite as

The potent in vitro antioxidant ellagitannins from pomegranate juice are metabolised into bioavailable but poor antioxidant hydroxy–6H–dibenzopyran–6– one derivatives by the colonic microflora of healthy humans

  • B.  Cerdá
  • J. C. Espín
  • S. Parra
  • P. Martínez
  • F. A. Tomás-BarberánEmail author



The antiatherogenic activity of pomegranate juice has been attributed to its antioxidant polyphenols. The most potent in vitro antioxidant polyphenol from this juice is the ellagitannin punicalagin. However, the bioavailability of ellagitannins, including punicalagin, has not been previously described in humans.

Aim of the study

The present work aims to evaluate, in healthy humans, the bioavailability and metabolism of pomegranate juice ellagitannins, to assess their effect on several blood parameters (including cardiovascular risk disease markers) and to compare the antioxidant activity of punicalagin with that of the in vivo generated metabolites.


Six healthy subjects (four men and two women) consumed 1 L of pomegranate juice daily (5.58 g/L polyphenols, including 4.37 g/L punicalagin isomers) for 5 days. The polyphenols and the in vivo generated metabolites were measured by HPLC–DAD–MS–MS. Fourteen haematological and twenty serobiochemical parameters including LDL, HDL and VLDL as well as cholesterol and triglycerides in each lipoprotein were evaluated. In vitro antioxidant activity of plasma (ABTS and FRAP assays) and urine (ABTS and DPPH) were determined.


Neither punicalagin nor ellagic acid present in the juice were detected in both plasma and urine. Three microbial ellagitannin-derived metabolites were detected: 3,8–dihydroxy–6H–dibenzo[b,d] pyran–6–one glucuronide, an unidentified aglycone (tentatively, trihydroxy–6H–dibenzo[b,d]pyran–6–one) and hydroxy–6–Hdibenzo[ b,d]pyran–6–one glucuronide. These metabolites could reach up to 18.6 µM in plasma, although a large inter–individual variability was observed. In urine, the same metabolites and their corresponding aglycones became evident after 1 day of juice consumption. Total urine excretion of metabolites ranged from 0.7 to 52.7% regarding the ingested punicalagin. No relevant effect was observed on any blood parameter. The metabolites did not show significant antioxidant activity compared to punicalagin from pomegranate juice.


The potential systemic biological effects of pomegranate juice ingestion should be attributed to the colonic microflora metabolites rather than to the polyphenols present in the juice.

Key words

bioavailability antioxidant metabolism hydroxy–6H–dibenzo [b,d] pyran–6–one ellagitannin polyphenol punicalagin microbial metabolites pomegranate juice Punica granatum 


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  1. 1.
    Madihassan S (1984) Outline of the beginnings of alchemy and its antecedents. Am J Chinese Med 12:32–42Google Scholar
  2. 2.
    Gil MI, Tomás-Barberán FA, Hess- Pierce B, Holcroft DM, Kader AA (2000) Antioxidant acivity of pomegranate juice and its relationship with phenolic composition and processing. J Agric Food Chem 48:4581–4589PubMedGoogle Scholar
  3. 3.
    Aviram M, Dornfeld L (2001) Pomegranate juice consumption inhibits serum angiotensin converting enzyme activity and reduces systolic blood pressure. Atherosclerosis 158:195–198CrossRefPubMedGoogle Scholar
  4. 4.
    Aviram M, Dornfeld L, Rosenblat M, Volkova N, Kaplan M, Coleman R, Hayek T, Presser D, Fuhrman B (2000) Pomegranate juice consumption reduces oxidative stress, atherogenic modifications to LDL, and platelet aggregation: studies in humans and atherosclerotic apolipoprotein E-deficient mice. Am J Clin Nutr 71:1062–1076PubMedGoogle Scholar
  5. 5.
    Clifford MN, Scalbert A (2000) Elagitannins- nature, occurrence and dietary burden. J Sci Food Agric 80:118–125Google Scholar
  6. 6.
    Tomás-Barberán FA, Clifford MN (2000) Dietary hydroxybenzoic acid derivatives- nature, occurrence and dietary burden. J Sci Food Agric 80:1024–1032CrossRefGoogle Scholar
  7. 7.
    Doyle B,Griffiths LA (1980) The metabolism of ellagic acid in the rat. Xenobiotica 10:247–256PubMedGoogle Scholar
  8. 8.
    Smart RC, Huant MT, Chang RL, Sayer JM, Jerina DM, Conney AH (1986) Disposition of the naturally occurring antimutagenic plant phenol, ellagic acid, and its synthetic derivatives 3-O-decylellagic acid and 3,3’-di-O-methylellagic acid in mice. Carcinogenesis 7:1663–1667PubMedGoogle Scholar
  9. 9.
    Teel RW, Martin RM (1988) Disposition of the plant phenol ellagic acid in the mouse following oral administration by gavage. Xenobiotica 18:397–405PubMedGoogle Scholar
  10. 10.
    Cerdá B, Llorach R, Cerón JJ, Espín JC, Tomás-Barberán FA (2003) Evaluation of the bioavailability and metabolism in the rat of punicalagin, and antioxidant polyphenol from pomegranate juice. Eur J Nutr 42:18–28CrossRefPubMedGoogle Scholar
  11. 11.
    Tomás-Barberán FA, Blázquez MA, García-Viguera C, Ferreres F, Tomás- Lorente FA (1992) Comparative study of different Amberlite XAD resins in flavonoid analysis. Phytochem Anal 3: 78–81Google Scholar
  12. 12.
    Larrosa M, Espín JC, Tomás-Barberán FA (2003) Antioxidant capacity of tomato juice functionalized with enzymatically synthesized hydroxytyrosol. J Sci Food Agric 83:658–666Google Scholar
  13. 13.
    Rice-Evans CA, Miller JN (1994) Total antioxidant status in plasma and body fluids. Meth Enzymol 234:279–293PubMedGoogle Scholar
  14. 14.
    Espín JC, Wichers HJ (2000) Study of the oxidation of resveratrol catalyzed by polyphenol oxidase. Effect of polyphenol oxidase on the antiradical capacity of resveratrol. J Food Biochem 24:225–250Google Scholar
  15. 15.
    Llorach R, Espín JC, Tomás-Barberán FA, Ferreres F (2003) Valorization of cauliflower (Brassica oleracea L var. botrytis) by-products as a source of antioxidant phenolics. J Agric Food Chem 51:2181–2187CrossRefPubMedGoogle Scholar
  16. 16.
    Bachorik PS, Ross JW (1995) National cholesterol education program recommendations for measurement of lowdensity lipoprotein cholesterol: executive summary. Clin Chem 41:1414–1420PubMedGoogle Scholar
  17. 17.
    Wonnacott TH, Wonnacott RJ (1990) Introductory statistics. John Wiley & Sons,New YorkGoogle Scholar
  18. 18.
    Daniel EM, Ratnayake S, Kinstle T, Soner GD (1991) The effects of pH and rat intestinal contents on the liberation of ellagic acid from purified and crude ellagitannins. J Nat Prod 54:946–952PubMedGoogle Scholar
  19. 19.
    Fuhr U,Kummert AL (1995) The fate of naringin in humans: a key to grapefruit juice-drug interactions? Clin Pharmacol Ther 58:365–373PubMedGoogle Scholar
  20. 20.
    Virgili F, Pagana G, Bourne L, Rimbach G, Natella F, Rice-Evans C, Packer L (2000) Ferulic acid excretion as a marker of consumption of a French maritime pine (Pinus maritima) bark extract. Free Radic Biol Med 28:1249–1256CrossRefPubMedGoogle Scholar
  21. 21.
    Ferreres F, Tomás-Lorente F, Tomás- Barberán FA (1989) Current trends in plant flavonoid analysis. In: Atta-ur- Rhaman (ed) Studies in Natural Products Chemistry. Elsevier, Amsterdam, pp 655–656Google Scholar
  22. 22.
    Jeong SJ, Kim NY, Kim DH, Kang TH, Ahn NH, Miyamoto T, Higuchi R, Kim YC (2000) Hyaluronidase inhibitory active 6H-dibenzo[b,d]pyran-6-ones from the feces of Trogopterus xanthippes. Planta Med 66:76–77PubMedGoogle Scholar
  23. 23.
    Milbury P, Cao G, Prior RL, Blumberg J (2002) Bioavailability of elderberry anthocyanins. Mech Ageing Dev 123:997–1006CrossRefPubMedGoogle Scholar
  24. 24.
    Cao G, Muccitelli HU, Sánchez-Moreno C, Prior RL (2001) Anthocyanins are absorbed in glycated forms in elderly women: a pharmacokinetic study. Am J Clin Nutr 73:920–926PubMedGoogle Scholar
  25. 25.
    Scalbert A, Williamson G (2000) Dietary intake and bioavailability of polyphenols. J Nutr 130:2073–2085Google Scholar
  26. 26.
    Buckingham J (1992) Dictionary of Natural Products. Chapman & Hall, LondonGoogle Scholar
  27. 27.
    Cerdá B, Tomás-Barberán FA, Espín JC (2003) The repeated oral administration of high doses of the pomegranate ellagitannin punicalagin to rats for 37 days is not toxic. J Agric Food Chem 51:3493–3501CrossRefPubMedGoogle Scholar
  28. 28.
    Bernays EA, Cooper-Driver G, Bilgener M (1989) Herbivores and plant tannins. Adv Ecol Res 19:263–301Google Scholar

Copyright information

© Steinkopff Verlag 2004

Authors and Affiliations

  • B.  Cerdá
    • 1
  • J. C. Espín
    • 1
  • S. Parra
    • 2
  • P. Martínez
    • 2
  • F. A. Tomás-Barberán
    • 1
    Email author
  1. 1.Research Group on Quality, Safety and Bioactivity of Plant FoodsDept. Food Science and Technology, CEBAS-CSICEspinardo (Murcia)Spain
  2. 2.Clinical Analysis Service Laboratory of BiochemistryVirgen de La Arrixaca University HospitalMurciaSpain

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