Antioxidant properties of polyphenol-rich cocoa products industrially processed (original) (raw)

Abstract

Fermentation and roasting are the main causes of polyphenol degradation during the process for obtaining cocoa products. In the present study, a process for obtaining polyphenol-rich cocoa products on an industrial scale is described. The process avoids the fermentation and roasting steps and includes a step for the inactivation of the enzyme Polyphenol Oxidase (PPO), which helps preserve the polyphenol content present in the raw cocoa bean. In addition, our study evaluates the antioxidant capacity and characterizes the flavonoid profile of the polyphenol-rich cocoa products obtained from the natural polyphenolrich cocoa cake. Using different protocols, we have obtained three cocoa extracts with high polyphenol content, namely extracts A (167 mg/g), B (374 mg/g) and C (787 mg/g). The scavenging capacity of the extracts was measured as their ability to bleach the stable radicals DPPH Å and ABTS Å+ while their antioxidant effect was evaluated with the FRAP assay. The results for A, B and C in the DPPH test expressed as Trolox equivalent (lmol)/mg dry weight of extract were 0.2, 1.4 and 3.0, respectively; in the ABTS test the results were 1.0, 4.7 and 9.8. The antioxidant capacity expressed as ascorbic acid equivalent (lmol)/mg dry weight of each product were 17.2, 76.1 and 207.7, respectively. The scavenging properties of cocoa powder against the superoxide anion, H 2 O 2 , HClO, and peroxynitrite were also determined. The IC 50 (lg/mL) values in the hypoxanthine/xanthine oxidase test were 77.5, 12.3 and 10.3, for A, B and C, respectively, while as an HOCl scavenger the IC 50 (lg/mL) values were 225.4, 73.2 and 21.5. As a peroxynitrite anion scavenger, only extract C had a relevant effect, with IC 50 (lg/mL) values of 76.1 or 110.0 in the absence or presence of bicarbonate. None of the extracts tested showed activity in the hydrogen peroxide test, but B and C significantly increased the deoxyribose degradation in the absence of ascorbate. Likewise, none of the extracts inhibited the ferrous or copper chelating activity at 100 lg/mL, but they inhibited the lipid peroxidation in brain homogenates and human plasma through non-enzymatic generation systems, with extract C giving the best IC 50 (lg/mL) values: 17.4 and 8.1 against lipid peroxidation in brain homogenates and human plasma, respectively. In conclusion, if the extractive protocol is well characterized, defined and optimized, cocoa could constitute a source of polyphenols for enriching foods, nutraceuticals and alimentary supplements.

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References (76)

  1. Adamson, G. E., Lazarus, S. A., Mitchell, A. E., Prior, R. L., Cao, G., Jacobs, P. H., et al. (1999). HPLC method for the quantification of procyanidins in cocoa and chocolate samples and correlation to total antioxidant capacity. Journal of Agricultural and Food Chemistry, 47, 4184-4188.
  2. Armstrong, D. (1998). Preface. In D. Armstrong (Ed.), Free radicals and antioxidant protocols. Totowa, New Jersey: Humana Press.
  3. Andrés-Lacueva, C., Lamuela-Raventós, R. M., & Jáuregui, O. (2000). An LC method for the analysis of cocoa phenolics. LC GC Europe, 902-905.
  4. Aruoma, O. I. (2003). Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. Mutation Research, 523-524, 9-20.
  5. Aruoma, O. I., Bahorun, T., & Jen, L. S. (2003). Neuroprotection by bioactive components in medicinal and food plant extracts. Mutation Research, 544, 203-215.
  6. Aviram, M. (1995). Oxidative modification of low density lipoprotein and its relation to atherosclerosis. Israel Journal of Medical Sciences, 31, 241-249.
  7. Azad, N., Rojanasakul, Y., & Vallyathan, V. (2008). Inflammation and lung cancer: Roles of reactive oxygen/nitrogen species. Journal of Toxicology and Environmental Health. Part B, Critical Reviews, 11, 1-15.
  8. Balavoine, G. G., & Geletii, Y. V. (1999). Peroxynitrite scavenging by different antioxidants. Part I: Convenient assay. Nitric Oxide, 3, 40-54.
  9. Benzie, I. F. F., & Strain, J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of ''antioxidant power": The FRAP assay. Analytical Biochemistry, 239, 70-76.
  10. Bradbury, A. G. W., & Kschonek, E. M. (2008). Cocoa beans with reduce polyphenol oxidase activity and high polyphenol content. EP2005835 (A1). USA: Kraft Foods R&D Inc.
  11. Burkitt, M. J., & Gilbert, B. C. (1990). Model studies of the iron-catalysed Haber- Weiss cycle and the ascorbate-driven Fenton reaction. Free Radical Research Communications, 10, 265-280.
  12. Cavin, A., Hostettmann, K., Dyatmyko, W., & Potterat, O. (1998). Antioxidant and lipophilic constituents of Tinospora crispa. Planta Medica, 64, 393-396.
  13. Cienfuegos-Jovellanos, E., Pasamar, M. A., Fritz, J., Arcos, J., Ramón, D., & Castilla, Y. (2007). Proceso de obtención de cacao en polvo rico en polifenoles y bajo contenido en materia grasa, y cacao así obtenido. Patent Cooperation Treaty (PCT) WO 2007/096449A1. Spain: Natraceutical Industrial.
  14. Cienfuegos-Jovellanos, E., Quiñones, M. M., Muguerza, B., Moulay, L., Miguel, M., & Aleixandre, A. (2009). Antihypertensive effect of a polyphenol-rich cocoa powder industrially processed to preserve the original flavonoids of the cocoa beans. Journal of Agricultural and Food Chemistry, 57, 6156-6162.
  15. Cooper, K. A., Donovan, J. L., Waterhouse, A. L., & Williamson, G. (2009). Cocoa and health: A decade of research. British Journal of Nutrition, 99, 1-11.
  16. Dillon, S. A., Burmi, R. S., Lowe, G. M., Billington, D., & Rahman, K. (2003). Antioxidant properties of aged garlic extract: An in vitro study incorporating human low density lipoprotein. Life Sciences, 2003(72), 1583-1594.
  17. Federico, A., Morgillo, F., Tuccillo, C., Ciardiello, F., & Loguercio, C. (2007). Chronic inflammation and oxidative stress in human carcinogenesis. International Journal of Cancer, 121, 2381-2386.
  18. Fisher, N. D. L., Hughes, M., Gerhard-Herman, M., & Hollenberg, N. K. (2003). Flavonoid-rich cocoa induces nitric-oxide-dependent vasodilation in healthy humans. Journal of Hypertension, 21, 2281-2286.
  19. Flammer, A. J., Hermann, F., Sudano, I., Spieker, L., Hermann, M., Cooper, K. A., et al. (2007). Dark chocolate improves coronary vasomotion and reduces platelet reactivity. Circulation, 116, 2376-2382.
  20. Freeman, B. A., & Crapo, J. D. (1982). Biology of disease: Free radicals and tissue injury. Laboratory Investigation, 47, 412-426.
  21. Gaut, J. P., & Heinecke, J. W. (2001). Mechanisms for oxidizing low-density lipoprotein. Insights from patterns of oxidation products in the artery wall and from mouse models of atherosclerosis. Trends in Cardiovascular Medicine, 11, 103-112.
  22. Góngora, L., Giner, R. M., Máñez, S., Recio, M. C., Schinella, G., & Ríos, J. L. (2002). Effects of caffeoyl conjugates of isoprenyl-hydroquinone glucoside and quinic acid on leukocyte function. Life Sciences, 71, 2995-3004.
  23. Halliwell, B. (2009). The wanderings of a free radical. Free Radicals in Biology and Medicine, 46, 531-542.
  24. Halliwell, B., Gutteridge, J. M. C., & Aruoma, O. I. (1987). The deoxyribose method: Simple ''test-tube" assay for determination of rate constants for reactions of hydroxyl radicals. Analycal Biochemistry, 165, 215-219.
  25. Hatano, T., Miyatake, H., Natsume, M., Osakabe, N., Takizawa, T., Ito, H., et al. (2002). Proanthocyanidin glycosides and related polyphenols from cacao liquor and their antioxidant effects. Phytochemistry, 59, 749-758.
  26. Heinrich, M., & Prieto, J. M. (2008). Diet and healthy ageing 2100: Will we globalise local knowledge systems? Ageing Research Review, 7, 249-274.
  27. Heiss, C., Kleinbongard, P., Dejam, A., Perre, S., Schroeter, H., Sies, H., et al. (2005). Acute consumption of flavanol-rich cocoa and the reversal of endothelial dysfunction in smokers. Journal of the American College of Cardiology, 46, 1276-1283.
  28. Holt, R. R., Lazarus, S. A., Sullards, M. C., Zhu, Q. Y., Schramm, D. D., Hammerstone, J. F., et al. (2002). Procyanidin dimmer B2 [epicatechin-(4b-8)-epicatechin] in human plasma after consumption of a flavanol-rich cocoa. American Journal of Clinical Nutrition, 76, 798-804.
  29. Horsley, E. T., Burkitt, M. J., Jones, C. M., Patterson, R. A., Harris, L. K., Moss, N. J., et al. (2007). Mechanism of the antioxidant to pro-oxidant switch in the behavior of dehydroascorbate during LDL oxidation by copper(II) ions. Archives of Biochemistry and Biophysics, 465, 303-314.
  30. Huber, A., Stuchbury, G., Bürkle, A., Burnell, J., & Münch, G. (2006). Neuroprotective therapies for Alzheimer's disease. Current Pharmaceutical Design, 12, 705-717.
  31. Jarjour, N. N., & Calhoun, W. J. (1994). Enhanced production of oxygen radicals in asthma. The Journal of Laboratory and Clinical Medicine, 123, 131-136.
  32. Jie, G., Lin, Z., Zhang, L., Lv, H., He, P., & Zhao, B. (2006). Free radical scavenging effect of Pu-erh tea extracts and their protective effect on oxidative damage in human fibroblast cells. Journal of Agricultural and Food Chemistry, 54, 8058-8064.
  33. Juan, M. E., Wenzel, U., Ruiz-Gutiérrez, V., Daniel, H., & Planas, J. M. (2006). Olive fruit extracts inhibit proliferation and induce apoptosis in HT-29 human colon cancer cells. Journal of Nutrition, 136, 2553-2557.
  34. Kaplan, M., Mutlu, E. A., Benson, M., Fields, J. Z., Banan, A., & Keshavarzian, A. (2007). Use of herbal preparations in the treatment of oxidant-mediated inflammatory disorders. Complementary Therapies in Medicine, 15, 207-216.
  35. Kattenberg, H. R., & Willemsen, J. H. A. (2003). The manufacture of cocoa polyphenols as healthy supplements in food products. IFIS Journal. Food Science -Central 3. <http://www.foodsciencecentral.com/library.html#ifis/ 12553>.
  36. Kennetta, E. C., & Davies, M. J. (2007). Degradation of matrix glycosaminoglycans by peroxynitrite/peroxynitrous acid: Evidence for a hydroxyl-radical-like mechanism. Free Radical in Biology and Medicine, 42, 1278-1289.
  37. Keshavarzian, A., Sedghi, S., Kanofsky, J., List, T., Robinson, C., Ibrahim, C., et al. (1992). Excessive production of reactive oxygen metabolites by inflamed colon: Analysis by chemiluminescence probe. Gastroenterology, 103, 177-185.
  38. Kim, H., & Keeney, P. G. (1984). (-)-Epicatechin content in fermented and unfermented cocoa beans. Journal of Food Sciences, 49, 1090-1092.
  39. Koppenol, W. H., Kissner, R., & Beckman, J. S. (1996). Syntheses of peroxynitrite: To go with the flow or on solid grounds? Methods in Enzymology, 269, 296-302.
  40. Lamuela-Raventós, R. M., Romero-Pérez, A. I., Andrés-Lacueva, C., & Tornero, A. (2005). Review: Health effects of cocoa flavonoids. Food Science and Technology International, 11, 159-176.
  41. Lee, K. W., Kim, Y. J., Lee, H. J., & Lee, C. Y. (2003). Cocoa has more phenolic phytochemicals and a higher antioxidant capacity than teas and red wine. Journal of Agricultural and Food Chemistry, 51, 7292-7295.
  42. Liu, F., & Ng, T. B. (2000). Antioxidative and free radical scavenging activities of selected medicinal herbs. Life Sciences, 66, 725-735.
  43. Lopes, G. K. B., Schulman, H. M., & Hermes-Lima, M. (1994). Polyphenol tannic acid inhibits hydroxyl radical formation from Fenton reaction by complexing ferrous ions. Biochimica and Biophysica Acta, 1472, 142-152.
  44. Montesinos, M. C., Úbeda, A., Terencio, M. C., Payá, M., & Alcaraz, M. J. (1995). Antioxidant profile of mono-and dihydroxylated flavone derivatives in free radical generating systems. Zeitschrift für Naturforschung, 50c, 552-560.
  45. Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65, 55-63.
  46. Motohashi, N., Kawase, M., Kurihara, T., Shirataki, Y., Kamata, K., Nakashima, H., et al. (1999). Relationship between radical intensity and biological activity of cacao husks extracts. Anticancer Research, 19, 1125-1130.
  47. Ng, T. B., Liu, F., & Wang, Z. T. (2000). Antioxidative activity of natural products from plants. Life Sciences, 66, 709-723.
  48. Odeh, M. (1997). New insights into the pathogenesis and treatment of rheumatoid arthritis. Clinical Immunology and Immunopathology, 83, 103-116.
  49. Pannala, A. S., Chan, T. S., O'Brien, J., & Rice-Evans, C. (2001). Flanonoid B-ring chemistry and antioxidant activity: Fast reaction kinetics. Biochemical and Biophysical Research Communications, 282, 1161-1168.
  50. Pasamar, M. A., Ibarra, A., Cienfuegos-Jovellanos, E., & Laghi, S. (2006). Proceso de obtención de extractos de cacao con elevado contenido de polifenoles. Patent Cooperation Treaty (PCT) PCT/ES 2006 000294, Natraceutical Industrial, Spain.
  51. Pitchumoni, S. S., & Doraiswamy, P. M. (1998). Current status of antioxidant therapy for Alzheimer's disease. Journal of the American Geriatrics Society, 46, 1566-1572.
  52. Rein, D., Paglieroni, T. G., Wun, T., Pearson, D. A., Schmitz, H. H., Gosselin, R., et al. (2000). Cocoa inhibits platelet activation and function. American Journal of Clinical Nutrition, 72, 30-35.
  53. Rein, D., Lotito, S., Holt, R. R., Keen, C. L., Schmitz, H. H., & Fraga, C. G. (2000). Epicatechin in human plasma: In vivo determination and effect of chocolate consumption on plasma oxidation status. Journal of Nutrition, 130, 2109S-21014S.
  54. Ren, W., Qiao, Z., Wang, H., Zhu, L., & Zhang, L. (2003). Flavonoids: Promising anticancer agents. Medical Research Reviews, 23, 519-534.
  55. Rios, L. Y., Bennet, R. N., Lazarus, S. A., Rémésy, C., Scalbert, A., & Williamson, G. (2002). Cocoa procyanidins are stable during gastric transit in humans. American Journal of Clinical Nutrition, 76, 1106-1110.
  56. Ruch, R. J., Cheng, S. J., & Klaunig, J. E. (1989). Prevention of cytotoxicity and inhibition of intercellular communication by antioxidant catechins isolated from Chinese green tea. Carcinogenesis, 6, 1003-1008.
  57. Sanbongi, C., Osakabe, N., Natsume, M., Takizawa, T., Gomi, S., & Osawa, T. (1998). Antioxidative polyphenols isolated from Theobroma cacao. Journal of Agricultural and Food Chemistry, 46, 454-457.
  58. Schlesier, K., Harwat, M., Bohm, V., & Bitsch, R. (2002). Assessment of antioxidant activity by using different in vitro methods. Free Radical Research, 36, 177-187.
  59. Schinella, G. R., Troiani, G., Dávila, V., de Buschiazzo, P. M., & Tournier, H. A. (2000). Antioxidant effects of an aqueous extract of Ilex paraguariensis. Biochemical and Biophysical Research Communication, 269, 357-360.
  60. Schinella, G. R., Tournier, H. A., Máñez, S., de Buschiazzo, P. M., Recio, M. C., & Ríos, J. L. (2007). Tiliroside and gnaphaliin inhibit human low density lipoprotein oxidation. Fitoterapia, 78, 1-6.
  61. Serafini, M., Bugianesi, R., Maiani, G., Valtuena, S., De Santis, S., & Crozier, A. (2003). Plasma antioxidants from chocolate. Nature, 424, 1013.
  62. Sheweita, S. A., Tilmisany, A. M., & Al-Sawaf, H. (2005). Mechanisms of male infertility: Role of antioxidants. Current Drug Metabolism, 6, 495-501.
  63. Sies, H., Schewe, T., Heiss, C., & Kelm, M. (2005). Cocoa polyphenols and inflammatory meditors. American Journal of Clinical Nutrition, 81, S304-S312.
  64. Singleton, V. L., & Rossi, J. A. (1965). Colorimetric of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16, 144-158.
  65. Surh, Y. (1999). Molecular mechanisms of chemopreventive effects of selected dietary and medicinal phenolic substances. Mutation Research, 428, 305-327.
  66. Taubert, D., Roesen, R., Lehmann, C., Jung, N., & Schömig, E. (2007). Effects of low habitual cocoa intake on blood pressure and bioactive nitric oxide. A randomize control trial. The Journal of the American Medical Association, 298, 49-60.
  67. Tomás-Barberán, F. A., Cienfuegos-Jovellanos, E., Marín, A., Muguerza, B., Gil- Izquierdo, A., Cerdá, B., et al. (2007). A new process to develop a cocoa powder with higher flavonoid monomer content and enhanced bioavailability in healthy humans. Journal of Agricultural and Food Chemistry, 55, 3926-3935.
  68. Victor, V. M., Rocha, M., Esplugues, J. & De la Fuente, M. (2005). Role of free radicals in sepsis: Antioxidant therapy. Current Pharmaceutical Design, 11, 3141-3158.
  69. Wang, J. Y., Wen, L. L., Huang, Y. N., Chen, Y. T., & Ku, M. C. (2006). Dual effects of antioxidants in neurodegeneration: Direct neuroprotection against oxidative stress and indirect protection via suppression of glia-mediated inflammation. Current Pharmaceutical Design, 12, 3521-3533.
  70. Wang-Polagruto, J. F., Villablanca, A. C., Polagruto, J. A., Lee, L., Holt, R. R., Schrader, H. R., et al. (2006). Chronic consumption of flavanol-rich cocoa improves endothelial function and decreases vascular cell adhesion molecule in hypercholesterolemic postmenopausal women. Journal of Cardiovascular Pharmacology, 47, S177-S186.
  71. Waterhouse, A. L., Shirley, J. R., & Donovan, J. L. (1996). Antioxidants in chocolate. The Lancet, 348, 834.
  72. Weiss, S. J., Klein, R., Slivka, A., & Wei, M. (1982). Chlorination of taurine by human neutrophils. Evidence for hypochlorous acid generation. Journal of Clinical Investigation, 70, 598-607.
  73. Witztum, J. L., & Steinberg, D. (1991). Role of oxidized low density lipoprotein in atherogenesis. Journal of Clinical Investigation, 88, 1785-1792.
  74. Wollgast, J., & Anklam, E. (2000). Polyphenols in chocolate: Is there a contribution to human health? Food Research International, 33, 449-459.
  75. Yasui, K., & Baba, A. (2006). Therapeutic potential of superoxide dismutase (SOD) for resolution of inflammation. Inflammation Research, 55, 359-363.
  76. Yen, G. C., Duh, P. D., & Chuang, D. Y. (2000). Antioxidant activity of anthraquinones and anthorne. Food Chemistry, 70, 437-441.