Analytical and Bioanalytical Chemistry

, Volume 394, Issue 6, pp 1545–1556

Epicatechin, procyanidins, and phenolic microbial metabolites after cocoa intake in humans and rats

  • Mireia Urpi-Sarda
  • Maria Monagas
  • Nasiruddin Khan
  • Rosa M. Lamuela-Raventos
  • Celestino Santos-Buelga
  • Emilio Sacanella
  • Margarida Castell
  • Joan Permanyer
  • Cristina Andres-Lacueva
Original Paper

Abstract

Proanthocyanidins, flavonoids exhibiting cardiovascular protection, constitute a major fraction of the flavonoid ingested in the human diet. Although they are poorly absorbed, they are metabolized by the intestinal microbiota into various phenolic acids. An analytical method, based on an optimized 96-well plate solid-phase extraction (SPE) procedure and liquid chromatography tandem mass spectrometry (SPE-LC-MS/MS) for the analysis of 19 phenolic microbial metabolites and monomeric and dimeric flavanols in urine samples, was developed and validated. Human urine samples were obtained before and after ingestion of an acute consumption of 40 g of soluble cocoa powder and rat urines before and after the prolonged administration (2 weeks) of different diets composed of natural cocoa powder. The mean recovery of analytes using the new SPE-LC-MS/MS method ranged from 87% to 109%. Accuracy ranged from 87.5% to 113.8%, and precision met acceptance criteria (<15% relative standard deviation). Procyanidin B2 has been detected and quantified for the first time in human and rat urine after cocoa consumption. Changes in human and rat urinary levels of microbial phenolic acids and flavanols were in the range of 0.001–59.43 nmol/mg creatinine and of 0.004–181.56 nmol/mg creatinine, respectively. Major advantages of the method developed include reduction of laboratory work in the sample preparation step by the use of 96-well SPE plates and the sensitive measurement of a large number of metabolites in a very short run time, which makes it ideal for use in epidemiological studies.

Keywords

Cocoa Epicatechin Procyanidin Microbial metabolites LC-MS/MS 

References

  1. 1.
    Aron PM, Kennedy JA (2008) Mol Nutr Food Res 52:79–104CrossRefGoogle Scholar
  2. 2.
    Manach C, Williamson G, Morand C, Scalbert A, Remesy C (2005) Am J Clin Nutr 81:230–242Google Scholar
  3. 3.
    Deprez S, Brezillon C, Rabot S, Philippe C, Mila I, Lapierre C, Scalbert A (2000) J Nutr 130:2733–2738Google Scholar
  4. 4.
    Rechner AR, Kroner C (2005) Thromb Res 116:327–334CrossRefGoogle Scholar
  5. 5.
    Gao K, Xu A, Krul C, Venema K, Liu Y, Niu Y, Lu J, Bensoussan L, Seeram NP, Heber D, Henning SM (2006) J Nutr 136:52–57Google Scholar
  6. 6.
    Mennen LI, Sapinho D, Ito H, Bertrais S, Galan P, Hercberg S, Scalbert A (2006) Br J Nutr 96:191–198CrossRefGoogle Scholar
  7. 7.
    Spencer JP, Abd El Mohsen MM, Minihane AM, Mathers JC (2008) Br J Nutr 99:12–22Google Scholar
  8. 8.
    Gonthier MP, Cheynier V, Donovan JL, Manach C, Morand C, Mila I, Lapierre C, Remesy C, Scalbert A (2003) J Nutr 133:461–467Google Scholar
  9. 9.
    Gonthier MP, Donovan JL, Texier O, Felgines C, Remesy C, Scalbert A (2003) Free Radic Biol Med 35:837–844CrossRefGoogle Scholar
  10. 10.
    Barnes S, Prasain JK, Wang CC, Moore DR (2006) Life Sci 78:2054–2059CrossRefGoogle Scholar
  11. 11.
    Urpi-Sarda M, Zamora-Ros R, Lamuela-Raventos R, Cherubini A, Jauregui O, de la Torre R, Covas MI, Estruch R, Jaeger W, Andres-Lacueva C (2007) Clin Chem 53:292–299CrossRefGoogle Scholar
  12. 12.
    Urpi-Sarda M, Jauregui O, Lamuela-Raventos RM, Jaeger W, Miksits M, Covas MI, Andres-Lacueva C (2005) Anal Chem 77:3149–3155CrossRefGoogle Scholar
  13. 13.
    Gonthier MP, Rios LY, Verny M, Remesy C, Scalbert A (2003) J Chromatogr B Analyt Technol Biomed Life Sci 789:247–255CrossRefGoogle Scholar
  14. 14.
    Ito H, Gonthier MP, Manach C, Morand C, Mennen L, Remesy C, Scalbert A (2005) Br J Nutr 94:500–509CrossRefGoogle Scholar
  15. 15.
    Rios LY, Gonthier MP, Remesy C, Mila I, Lapierre C, Lazarus SA, Williamson G, Scalbert A (2003) Am J Clin Nutr 77:912–918Google Scholar
  16. 16.
    Roura E, Almajano MP, Bilbao ML, Andres-Lacueva C, Estruch R, Lamuela-Raventos RM (2007) Free Radic Res 41:943–949CrossRefGoogle Scholar
  17. 17.
    Andres-Lacueva C, Lamuela-Raventos RM (2000) LC-GC Eur.902–905Google Scholar
  18. 18.
    Roura E, Andres-Lacueva C, Estruch R, Lamuela-Raventos RM (2006) Clin Chem 52:749–752CrossRefGoogle Scholar
  19. 19.
    U.S.Department of Agriculture. USDA Database for the Proanthocyanidin Content of Selected Foods. http://www.nal.usda.gov/fnic/foodcomp.2004
  20. 20.
    Ramiro-Puig E, Urpi-Sarda M, Perez-Cano FJ, Franch A, Castellote C, Andres-Lacueva C, Izquierdo-Pulido M, Castell M (2007) J Agric Food Chem 55:6431–6438CrossRefGoogle Scholar
  21. 21.
    Miro-Casas E, Farre AM, Covas MI, Rodriguez JO, Menoyo CE, Lamuela Raventos RM, de la Torre R (2001) Anal Biochem 294:63–72CrossRefGoogle Scholar
  22. 22.
    U.S.Department of Health and Human Services, Food and Drug Administration. Guidance for Industry. Bioanalytical Method Validation; May 2001Google Scholar
  23. 23.
    Lee MJ, Maliakal P, Chen L, Meng X, Bondoc FY, Prabhu S, Lambert G, Mohr S, Yang CS (2002) Cancer Epidemiol. Biomarkers Prev 11:1025–1032Google Scholar
  24. 24.
    Li C, Lee MJ, Sheng SQ, Meng XF, Prabhu S, Winnik B, Huang BM, Chung JY, Yan SQ, Ho CT, Yang CS (2000) Chem Res Toxicol 13:177–184CrossRefGoogle Scholar
  25. 25.
    Andres-Lacueva C, Monagas M, Khan N, Izquierdo-Pulido M, Urpi-Sarda M, Permanyer J, Lamuela-Raventos RM (2008) J Agric Food Chem 56:3111–3117CrossRefGoogle Scholar
  26. 26.
    Holt RR, Lazarus SA, Sullards MC, Zhu QY, Schramm DD, Hammerstone JF, Fraga CG, Schmitz HH, Keen CL (2002) Am J Clin Nutr 76:798–804Google Scholar
  27. 27.
    Sano A, Yamakoshi J, Tokutake S, Tobe K, Kubota Y, Kikuchi M (2003) Biosci Biotechnol Biochem 67:1140–1143CrossRefGoogle Scholar
  28. 28.
    Baba S, Osakabe N, Natsume M, Terao J (2002) Free Radical Biology Medicine 33:142–148CrossRefGoogle Scholar
  29. 29.
    Shoji T, Masumoto S, Moriichi N, Akiyama H, Kanda T, Ohtake Y, Goda Y (2006) J Agric Food Chem 54:884–892CrossRefGoogle Scholar
  30. 30.
    Donovan JL, Manach C, Rios L, Morand C, Scalbert A, Remesy C (2002) Br J Nutr 87:299–306CrossRefGoogle Scholar
  31. 31.
    Tsang C, Auger C, Mullen W, Bornet A, Rouanet JM, Crozier A, Teissedre P (2005) Br J Nutr 94:170–181CrossRefGoogle Scholar
  32. 32.
    Prasain JK, Peng N, Dai Y, Moore R, Arabshahi A, Wilson L, Barnes S, Michael WJ, Kim H, Watts RL (2009) Phytomedicine 16:233–243Google Scholar
  33. 33.
    Waffo-Teguo P, Hawthorne ME, Cuendet M, Merillon JM, Kinghorn AD, Pezzuto JM, Mehta RG (2001) Nutr. Cancer 40:173–179CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Mireia Urpi-Sarda
    • 1
  • Maria Monagas
    • 2
  • Nasiruddin Khan
    • 1
  • Rosa M. Lamuela-Raventos
    • 1
  • Celestino Santos-Buelga
    • 3
  • Emilio Sacanella
    • 2
  • Margarida Castell
    • 4
  • Joan Permanyer
    • 1
  • Cristina Andres-Lacueva
    • 1
  1. 1.Nutrition and Food Science Department, XaRTA. INSA. Faculty of PharmacyUniversity of BarcelonaBarcelonaSpain
  2. 2.Department of Internal Medicine, Hospital Clínic. Institut d’Investigació Biomèdica August Pi i Sunyer (IDIBAPS)University of BarcelonaBarcelonaSpain
  3. 3.Laboratory of Nutrition and Bromatology, School of PharmacyUniversity of SalamancaSalamancaSpain
  4. 4.Department of Physiology, Faculty of PharmacyUniversity of BarcelonaBarcelonaSpain

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