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Bioactive Compounds from Theobroma cacao: Effect of Isolation and Safety Evaluation

Plant Foods for Human Nutrition volume 74, pages 40–46 (2019)Cite this article

Abstract

Plants, including most food and feed plants, produce a broad range of bioactive chemical compounds. Among these compounds, polyphenols are reported to provide beneficial effects as anti-carcinogenic, anti-atherogenic, anti-inflammatory, immune modulating, anti-microbial, vasodilatory and analgesic. Cocoa (Theobroma cacao), a major, economically important, international crop, has been related to several nutritional benefits, which have been associated with the phenolic fraction. The main subclass of flavonoids found in cocoa is flavanols, particularly (epi)catechins monomers, and their oligomers, also known as procyanidins. In this study, these compounds were isolated by different methodologies as solid phase extraction (SPE), semi-preparative high-performance liquid chromatography (HPLC) and membrane technologies to obtain different polyphenolic profiles by HPLC coupled to electrospray time-of-flight mass spectrometry (ESI-TOF-MS) and to test their cytotoxicity. Finally, different polyphenolic profiles were collected, where the combination of both semi-preparative HPLC and SPE technologies provided the most purified fractions. Filtration with membranes and SPE provide extracts with different composition depending on the pore size of membranes and on the solvent, respectively. In addition, the results of toxicity assay indicated low levels in all fractions.

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References

  1. Cádiz-Gurrea M de la L, Fernández-Arroyo S, Joven J et al (2015) Proanthocyanidins in agro-industrial by-products: health benefits. In: Motohashi N (ed) Occurrences, structure, biosynthesis, and health benefits based on their evidences of medicinal phytochemicals in vegetables and fruits, vol 3. Nova Science Publishers, New York, pp 63–114

  2. Yao LH, Jiang YM, Shi J et al (2004) Flavonoids in food and their health benefits. Plant Foods Hum Nutr 59:113–122. https://doi.org/10.1007/s11130-004-0049-7

    Article  CAS  PubMed  Google Scholar 

  3. Wollgast J, Anklam E (2000) Review on polyphenols in Theobroma cacao: changes in composition during the manufacture of chocolate and methodology for identification and quantification. Food Res Int 33:423–447. https://doi.org/10.1016/S0963-9969(00)00068-5

    Article  CAS  Google Scholar 

  4. Wollgast J, Pallaroni L, Agazzi ME, Anklam E (2001) Analysis of procyanidins in chocolate by reversed-phase high-performance liquid chromatography with electrospray ionisation mass spectrometric and tandem mass spectrometric detection. J Chromatogr A 926:211–220

    Article  CAS  PubMed  Google Scholar 

  5. Counet C, Collin S (2003) Effect of the number of flavanol units on the antioxidant activity of procyanidin fractions isolated from chocolate. J Agric Food Chem 51:6816–6822. https://doi.org/10.1021/jf030349g

    Article  CAS  PubMed  Google Scholar 

  6. Pérez-Jiménez J, Elena Díaz-Rubio M, Saura-Calixto F (2015) Contribution of macromolecular antioxidants to dietary antioxidant capacity: a study in the Spanish Mediterranean diet. Plant Foods Hum Nutr 70:365–370. https://doi.org/10.1007/s11130-015-0513-6

    Article  CAS  PubMed  Google Scholar 

  7. Andres-Lacueva C, Monagas M, Khan N, Izquierdo-Pulido M, Urpi-Sarda M, Permanyer J, Lamuela-Raventós RM (2008) Flavanol and flavonol contents of cocoa powder products: influence of the manufacturing process. J Agric Food Chem 56:3111–3117

    Article  CAS  PubMed  Google Scholar 

  8. Cádiz-Gurrea M de la L, Lozano-Sánchez J, del M Contreras-Gámez M et al (2014) Isolation, comprehensive characterization and antioxidant activities of Theobroma cacao extract. J Funct Foods 10:485–498. https://doi.org/10.1016/j.jff.2014.07.016

    Article  CAS  Google Scholar 

  9. Jiménez-Sánchez C, Olivares-Vicente M, Rodríguez-Pérez C, Herranz-López M, Lozano-Sánchez J, Segura-Carretero A, Fernández-Gutiérrez A, Encinar JA, Micol V (2017) AMPK modulatory activity of olive-tree leaves phenolic compounds: bioassay-guided isolation on adipocyte model and in silico approach. PLoS One 12:1–22. https://doi.org/10.1371/journal.pone.0173074

    Article  CAS  Google Scholar 

  10. Ortega N, Romero M-P, MacIà A et al (2008) Obtention and characterization of phenolic extracts from different cocoa sources. J Agric Food Chem 56:9621–9627

    Article  CAS  PubMed  Google Scholar 

  11. Monagas M, Gómez-Cordovés C, Bartolomé B, Laureano O, Ricardo da Silva JM (2003) Monomeric, oligomeric, and polymeric flavan-3-ol composition of wines and grapes from Vitis vinifera L. cv. Graciano, Tempranillo, and Cabernet Sauvignon. J Agric Food Chem 51:6475–6481. https://doi.org/10.1021/jf030325+

    Article  CAS  PubMed  Google Scholar 

  12. Pinelo M, Laurie VF, Waterhouse AL (2006) A simple method to separate red wine nonpolymeric and polymeric phenols by solid-phase extraction. J Agric Food Chem 54:2839–2844. https://doi.org/10.1021/jf052814a

    Article  CAS  PubMed  Google Scholar 

  13. Alonso García A, Cancho Grande B, Simal Gándara J (2004) Development of a rapid method based on solid-phase extraction and liquid chromatography with ultraviolet absorbance detection for the determination of polyphenols in alcohol-free beers. J Chromatogr A 1054:175–180. https://doi.org/10.1016/j.chroma.2004.07.092

    Article  CAS  PubMed  Google Scholar 

  14. Khemakhem I, Gargouri OD, Dhouib A, Ayadi MA, Bouaziz M (2017) Oleuropein rich extract from olive leaves by combining microfiltration, ultrafiltration and nanofiltration. Sep Purif Technol 172:310–317. https://doi.org/10.1016/j.seppur.2016.08.003

    Article  CAS  Google Scholar 

  15. Shao ZH, Hsu CW, Chang WT, Waypa GB, Li J, Li D, Li CQ, Anderson T, Qin Y, Schumacker PT, Becker LB, Hoek TLV (2006) Cytotoxicity induced by grape seed proanthocyanidins: role of nitric oxide. Cell Biol Toxicol 22:149–158. https://doi.org/10.1007/s10565-006-0052-x

    Article  CAS  PubMed  Google Scholar 

  16. Yamakoshi J, Saito M, Kataoka S, Kikuchi M (2002) Safety evaluation of proanthocyanidin-rich extract from grape seeds. Food Chem Toxicol 40:599–607. https://doi.org/10.1016/S0278-6915(02)00006-6

    Article  CAS  PubMed  Google Scholar 

  17. Lao W, Tan Y, Jin X, Xiao L, Kim JJY, Qu X (2015) Comparison of cytotoxicity and the anti-adipogenic effect of green tea polyphenols with epigallocatechin-3-gallate in 3T3-L1 preadipocytes. Am J Chin Med 43:1177–1190. https://doi.org/10.1142/S0192415X15500676

    Article  CAS  PubMed  Google Scholar 

  18. Sun B, Leandro MC, de Freitas V, Spranger MI (2006) Fractionation of red wine polyphenols by solid-phase extraction and liquid chromatography. J Chromatogr A 1128:27–38. https://doi.org/10.1016/j.chroma.2006.06.026

    Article  CAS  PubMed  Google Scholar 

  19. Butler LG, Price ML, Brotherton JE (1982) Vanillin assay for proanthocyanidins (condensed tannins): modification of the solvent for estimation of the degree of polymerization. J Agric Food Chem 30:1087–1089. https://doi.org/10.1021/jf00114a020

    Article  CAS  Google Scholar 

  20. Boulekbache-Makhlouf L, Medouni L, Medouni-Adrar S, Arkoub L, Madani K (2013) Effect of solvents extraction on phenolic content and antioxidant activity of the byproduct of eggplant. Ind Crop Prod 49:668–674. https://doi.org/10.1016/j.indcrop.2013.06.009

    Article  CAS  Google Scholar 

  21. Złotek U, Mikulska S, Nagajek M, Świeca M (2016) The effect of different solvents and number of extraction steps on the polyphenol content and antioxidant capacity of basil leaves (Ocimum basilicum L.) extracts. Saudi J Biol Sci 23:628–633. https://doi.org/10.1016/j.sjbs.2015.08.002

    Article  CAS  PubMed  Google Scholar 

  22. Bosso A, Guaita M, Petrozziello M (2016) Influence of solvents on the composition of condensed tannins in grape pomace seed extracts. Food Chem 207:162–169. https://doi.org/10.1016/j.foodchem.2016.03.084

    Article  CAS  PubMed  Google Scholar 

  23. Cassano A, Conidi C, Drioli E (2011) Comparison of the performance of UF membranes in olive mill wastewaters treatment. Water Res 45:3197–3204. https://doi.org/10.1016/j.watres.2011.03.041

    Article  CAS  PubMed  Google Scholar 

  24. Pinto PCR, Mota IF, Loureiro JM, Rodrigues AE (2014) Membrane performance and application of ultrafiltration and nanofiltration to ethanol/water extract of Eucalyptus bark. Sep Purif Technol 132:234–243. https://doi.org/10.1016/j.seppur.2014.04.042

    Article  CAS  Google Scholar 

  25. Hübsch Z, Van Zyl RL, Cock IE, Van Vuuren SF (2014) Interactive antimicrobial and toxicity profiles of conventional antimicrobials with southern african medicinal plants. S Afr J Bot 93:185–197. https://doi.org/10.1016/j.sajb.2014.04.005

    Article  Google Scholar 

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Acknowledgements

This work was supported by the project AGL2015-67995-C3-2-R (Spanish Ministry of Science and Innovation), as well as P11-CTS-7625 (Andalusian Regional Government Council of Innovation and Science). The European Social Fund (FSE) for the contract PTQ-13-06429.

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Authors and Affiliations

  1. Department of Analytical Chemistry, University of Granada, c/Fuentenueva s/n, 18071, Granada, Spain

    María de la Luz Cádiz-Gurrea & Antonio Segura-Carretero

  2. Research and Development of Functional Food Centre (CIDAF), PTS Granada, Avda. del Conocimiento 37, Edificio BioRegion, 18016, Granada, Spain

    María de la Luz Cádiz-Gurrea & Antonio Segura-Carretero

  3. Advanced NMR Methods and Metal-based Catalysts, University of Almería, Ctra. Sacramento s/n, La Cañada de San Urbano, 04120, Almería, Spain

    Ignacio Fernández de las Nieves & Luis Manuel Aguilera Saez

  4. Biomedical Research Unit. Medicine and Surgery Department, Rovira i Virgili University, 43201, Reus, Tarragona, Spain

    Salvador Fernández-Arroyo

  5. Institut National de la Santé et de la Recherche Médicale Unité 781, Université Paris Descartes-Sorbonne Paris Cité, Institut Imagine, Hôpital Necker-Enfants Malades, 75015, Paris, France

    Laurence Legeai-Mallet

  6. Laboratoire d’Electrochimie et Environnement, Ecole Nationale d’Ingénieurs de Sfax BP «1173» 3038, Université de Sfax, Sfax, Tunisie

    Mohamed Bouaziz

  7. Institut Supérieur de Biotechnologie de Sfax, Université de Sfax, BP «1175», 3038, Sfax, Tunisie

    Mohamed Bouaziz

Authors
  1. María de la Luz Cádiz-Gurrea
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  2. Ignacio Fernández de las Nieves
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  3. Luis Manuel Aguilera Saez
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  4. Salvador Fernández-Arroyo
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  7. Antonio Segura-Carretero
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Correspondence to María de la Luz Cádiz-Gurrea.

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de la Luz Cádiz-Gurrea, M., Fernández de las Nieves, I., Aguilera Saez, L.M. et al. Bioactive Compounds from Theobroma cacao: Effect of Isolation and Safety Evaluation. Plant Foods Hum Nutr 74, 40–46 (2019). https://doi.org/10.1007/s11130-018-0694-x

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  • DOI: https://doi.org/10.1007/s11130-018-0694-x

Keywords

  • Theobroma cacao
  • Food analysis
  • Polyphenols
  • Isolation
  • HPLC-ESI-TOF-MS