Publicado

2020-11-06

Extraction, separation, and evaluation of antioxidant effect of the different fractions of polyphenols from cocoa beans

Extracción, separación y evaluación del efecto antioxidante de las diferentes fracciones de polifenoles de los granos de cacao

Extração, Separação e Avaliação do Efeito Antioxidante das Diferentes Frações de Polifenóis de Grãos de Cacau

Palabras clave:

Flavonoids, antioxidant capacity, monomers, dimers, oligomers (en)
Flavonóides, capacidade antioxidante, monômeros, dímeros, oligômeros (pt)
Flavonoides, capacidad antioxidante, monómeros, dímeros, oligómeros (es)

Descargas

Autores/as

Polyphenol Extracts (PE) hold antioxidant properties, which might be related to positive effects on human health. It has been stated that PE, obtained from cocoa beans, contain fractions of flavan-3-ols with different degrees of polymerization (DP). However, it is unknown which of the fractions or their mixture drives the best antioxidant activity. This paper reports the study conducted to elucidate the role of each fraction (with different DP) on Antioxidant Capacity (AC). First, the process of extraction and separation of polyphenols’ fractions in cocoa beans was executed; afterward, AC was determined for each fraction individually and their combinations (monomers, dimers and oligomers). Solid-liquid extraction was made by using a 50% (v/v) ethanol solution and a mass:solvent ratio of 1:120 in an ultrasound bath. PE were separated into monomers, dimers, and oligomers by HPLC using a semipreparative column. The results obtained show that PE contain 95.35, 7.45, and 21.75 mg EE (epicatechin equivalents) / g ds of monomers, dimers, and oligomers, respectively. Finally, the AC of each fraction  was evaluated using a complete cubic model mix design. According to the results, the best AC was obtained for dimers. However, when monomers, dimers and oligomers were mixed, an antagonistic effect on AC was observed.

Los extractos polifenólicos (PE) tienen propiedades antioxidantes positivas para la salud humana. Se conoce que los PE obtenidos del cacao contienen fracciones de flavan-3-ols con diferentes grados de polimerización (DP). Sin embargo, se desconoce qué fracción o mezcla de ellos posee mayor capacidad antioxidante. Este trabajo se realizó con el fin de evaluar la capacidad antioxidante (AC) de cada fracción polifenólica (con diferentes DP) extraída del cacao. Primero, se realizó un proceso de extracción y separación de las fracciones de polifenoles en los granos de cacao. Luego, se determinó la AC para cada fracción individual y combinada (monómeros, dímeros y oligómeros). La extracción sólido-líquido se realizó utilizando una solución de etanol en agua al 50 % (v/v) y una relación masa:solvente de 1:120 en un baño de ultrasonido. El extracto PE se separó en monómeros, dímeros y oligómeros por HPLC usando una columna semipreparativa. Los resultados obtenidos muestran que el extracto PE contiene 95,35, 7,45 y 21,75 mg EE (equivalentes de epicatequina) / g ds de monómeros, dímeros y oligómeros, respectivamente. Finalmente, se evaluó la CA de cada fracción utilizando un diseño de mezcla de modelo cúbico completo. Según los resultados, se obtuvo una mayor AC para dímeros; sin embargo, se observó un efecto antagonista de la AC cuando se mezclan monómeros, dímeros y oligómeros.

Os extratos de polifenóis (PE) possuem propriedades antioxidantes, que podem estar relacionadas a efeitos positivos para a saúde humana. Foi afirmado que o PE, obtido a partir de grãos de cacau, contém frações de flavan-3-ols com diferentes graus de polimerização (DP). No entanto, não se sabe qual fração ou mistura deles impulsiona a melhor atividade antioxidante. Este artigo relata os achados de um estudo realizado com o objetivo de elucidar o papel de cada fração (com DP diferente) na capacidade antioxidante (AC). Inicialmente, o processo de extração e separação de frações de polifenóis 'em grãos de cacau é descrito. Posteriormente, é determinada a AC para cada fração isolada e combinada (monômeros, dímeros e oligômeros). A extração sólido-líquido foi realizada utilizando uma solução de etanol a 50% (v/v) e uma razão massa: solvente de 1: 120 em um banho de ultrassom. O PE foi separado em monômeros, dímeros e oligômeros por HPLC usando uma coluna semi-preparativa. Os resultados obtidos mostram que o PE contém 95,35, 7,45 e 21,75 mgEE/g ds de monômeros, dímeros e oligômeros, respectivamente. Finalmente, o efeito de cada fração na AC foi avaliado usando um modelo de mistura cúbica completo. De acordo com os resultados, a melhor AC foi obtida para os dímeros; no entanto, um efeito antagônico na AC foi observado quando monômeros, dímeros e oligômeros foram misturados.

Descargas

Los datos de descargas todavía no están disponibles.

Citas

G. A. R. Wood and R. A. Lass, “Cocoa”, 4th Ed. New York: The International Bank for Reconstruction, 1985.

J. A. Gil Quintero, “Estabilidad y actividad antioxidante de catequinas presentes en cacaos colombianos durante los procesos de pre e industrialización”, Masters Thesis in Pharmaceutical Sciences, Universidad de Antioquia, Medellín, Colombia, 2012.

J. Wollgast and E. Anklam, “Review on polyphenols in Theobroma cacao: Changes in composition during the manufacture of chocolate and methodology for identication® and quantication®”, Food Research International, vol. 33, n.o 6, pp. 423-447, 2000. DOI: 10.1016/S0963-9969(00)00068-5

J. Wollgast and E. Anklam, “Polyphenols in chocolate: is there a contribution to human health?”, Food Research International, vol. 33, n.o 6, pp. 449-459, 2000. DOI: 10.1016/S0963-9969(00)00069-7

H. Kim and P. G. Keeney, “ (-)-Epicatechin Content in Fermented and Unfermented Cocoa Beans”, Journal of Food Science, vol. 49, n.o 4, pp. 1090-1092, 1984. DOI: 10.1111/j.1365-2621.1984.tb10400.x

J. A. Vinson et al., “Chocolate Is a Powerful ex Vivo and in Vivo Antioxidant, an Antiatherosclerotic Agent in an Animal Model, and a Significant Contributor to Antioxidants in the European and American Diets”, J. Agric. Food. Chem., vol. 54, n.o 21, pp. 8071-8076, 2006. DOI: 10.1021/jf062175j

J. Pérez-Jiménez, V. Neveu, F. Vos, and A. Scalbert, “Identification of the 100 richest dietary sources of polyphenols: an application of the Phenol-Explorer database”, Eur. J. Clin. Nutr., vol. 64, n.o 3, pp. 112-120, 2010. DOI: 10.1038/ejcn.2010.221

J. Wollgast, “The contents and effects of polyphenols in chocolate: Qualitative and quantitative analyses of polyphenols in chocolate and chocolate raw products as well as evaluation of potential implications of chocolate consumption in human health”, Ph. D. dissertation, University of Gießen, Gießen, Germany, 2004.

A. Dube, K. Ng, J. A. Nicolazzo, and I. Larson, “Effective use of reducing agents and nanoparticle encapsulation in stabilizing catechins in alkaline solution”, Food Chemistry, vol. 122, n.o 3, pp. 662-667, 2010. DOI: 10.1016/j.foodchem.2010.03.027

M. Navarro, E. Arnaez, I. Moreira, A. Hurtado, D. Monge, and M. Monagas, “Polyphenolic Composition and Antioxidant Activity of Uncaria tomentosa Commercial Bark Products”, Antioxidants, vol. 8, n.o 9, p. 339, 2019. DOI: 10.3390/antiox8090339

K. S. Kealey, R. M. Snyder, and L. J. Romanczyk, “COCOA EXTRACTS PREPARED FROM COCOA SOLIDS HAVING HIGH COCOA POLYPHENOL CONTENTS”, US006737088B1, May 18, 2004.

R. M. Alonso-Salces, E. Korta, A. Barranco, L. A. Berrueta, B. Gallo, and F. Vicente, “Pressurized liquid extraction for the determination of polyphenols in apple”, J. Chromatogr. A, vol. 933, n.o 1-2, pp. 37-43, 2001. DOI: 10.1016/s0021-9673(01)01212-2. PMID: 11758745.

A. E. M. Abdalla, S. M. Darwish, E. H. E. Ayad, and R. M. El-Hamahmy, “Egyptian mango by-product 1. Compositional quality of mango seed kernel”, Food Chemistry, vol. 103, n.o 4, pp. 1134-1140, 2007. DOI: 10.1016/j.foodchem.2006.10.017

S. Mazor Jolić, I. Radojčić Redovniković, K. Marković, Đ. Ivanec Šipušić, and K. Delonga, “Changes of phenolic compounds and antioxidant capacity in cocoa beans processing”, Int. J. Food Sci. Tech., vol. 46, n.o 9, pp. 1793-1800, 2011. DOI: 10.1111/j.1365-2621.2011.02670.x

M. Tabernero, J. Serrano, and F. Saura-Calixto, “The antioxidant capacity of cocoa products: contribution to the Spanish diet”, Int. J. Food Sci. Tech., vol. 41, n.o s1, pp. 28-32, 2006. DOI: 10.1111/j.1365-2621.2006.01239.x

E. Dorta, M. González, M. G. Lobo, C. Sánchez-Moreno, and B. de Ancos, “Screening of phenolic compounds in by-product extracts from mangoes (Mangifera indica L.) by HPLC-ESI-QTOF-MS and multivariate analysis for use as a food ingredient”, Food Research International, vol. 57, pp. 51-60, 2014. DOI: 10.1016/j.foodres.2014.01.012

H. Kim et al., “Antioxidant and antiproliferative activities of mango (Mangifera indica L.) flesh and peel”, Food Chemistry, vol. 121, n.o 2, pp. 429-436, 2010. DOI: 10.1016/j.foodchem.2009.12.060

M. Navarro-Hoyos, D. Alvarado-Corella, I. Moreira-Gonzalez, E. Arnaez-Serrano, and M. Monagas-Juan, “Polyphenolic Composition and Antioxidant Activity of Aqueous and Ethanolic Extracts from Uncaria tomentosa Bark and Leaves”, Antioxidants, vol. 7, n.o 5, p. 65, 2018. DOI: 10.3390/antiox7050065

S. Toro-Uribe, L. J. López-Giraldo, and E. A. Decker, “Relationship between the Physiochemical Properties of Cocoa Procyanidins and Their Ability to Inhibit Lipid Oxidation in Liposomes”, J. Agric. Food Chem., vol. 66, n.o 17, pp. 4490-4502, 2018. DOI: 10.1021/acs.jafc.8b01074

S. Deprez, I. Mila, J.-F. Huneau, D. Tome, and A. Scalbert, “Transport of Proanthocyanidin Dimer, Trimer, and Polymer Across Monolayers of Human Intestinal Epithelial Caco-2 Cells”, Antioxidants & Redox Signaling, vol. 3, n.o 6, pp. 957-967, 2001. DOI: 10.1089/152308601317203503.

M. M. Rahman, T. Ichiyanagi, T. Komiyama, Y. Hatano, and T. Konishi, “Superoxide radical- and peroxynitrite-scavenging activity of anthocyanins; structure-activity relationship and their synergism”, Free Radic. Res., vol. 40, n.o 9, pp. 993-1002, 2006. DOI: 10.1080/10715760600815322.

D. Huang, B. Ou, M. Hampsch-Woodill, J. A. Flanagan, and R. L. Prior, “High-Throughput Assay of Oxygen Radical Absorbance Capacity (ORAC) Using a Multichannel Liquid Handling System Coupled with a Microplate Fluorescence Reader in 96-Well Format”, J. Agric. Food Chem., vol. 50, n.o 16, pp. 4437-4444, 2002. DOI: 10.1021/jf0201529

N. Martínez-Guerrero, “Aportes de la investigación de FEDECACAO – Fondo Nacional del Cacao al incremento de la productividad y reconocimiento de la calidad del cacao de Colombia”, FEDECACAO, 2016.

C. Álvarez, E. Pérez, and M. C. Lares, “ Physical-chemical characterization of fermented, dried and roasted cocoa beans cultivated in the region of Cuyagua, Aragua state”, Agronomía Tropical, vol. 57, n.o 4, pp. 249-256, 2007.

A. Vázquez-Ovando, F. Molina-Freaner, J. Nuñez-Farfán, D. Betancur-Ancona, and M. Salvador-Figueroa, “Classification of cacao beans (Theobroma cacao L.) of southern Mexico based on chemometric analysis with multivariate approach”, Eur. Food Res. Technol., vol. 240, pp. 1117-1128, 2015. DOI: 10.1007/s00217-015-2415-0

J. D. Peñaranda Herrera y L. J. Sierra Prada, “Evaluación de la actividad antioxidante, contenido de polifenoles y metilxantinas en materiales de cacao colombianos”, Thesis for undergraduate in Chemistry, Universidad Industrial de Santander, Bucaramanga, Colombia, 2013.

J. Alean, F. Chejne, and B. Rojano, “Degradation of polyphenols during the cocoa drying process”, Journal of Food Engineering, vol. 189, pp. 99-105, 2016. DOI: 10.1016/j.jfoodeng.2016.05.026

A. G. Yanzapanta Llambo, “Estudio de la variación de los contenidos de polifenoles totales, alcaloides y grasa en almendras de cacao fino de aroma en tres diferentes zonas de producción de la amazonía ecuatoriana”, Thesis for undergraduate in Food Engineering, Universidad Técnica de Ambato, Ambato, Ecuador, 2014.

S. Toro-Uribe, L. J. López-Giraldo, G. Alvarez-Rivera, E. Ibáñez, and M. Herrero, “Insight of Stability of Procyanidins in Free and Liposomal Form under an in Vitro Digestion Model: Study of Bioaccessibility, Kinetic Release Profile, Degradation, and Antioxidant Activity”, J. Agric. Food Chem., vol. 67, n.o 7, pp. 1990-2003, 2019. DOI: 10.1021/acs.jafc.9b00351.

V. Pedan, N. Fischer, and S. Rohn, “An online NP-HPLC-DPPH method for the determination of the antioxidant activity of condensed polyphenols in cocoa”, Food Research International, vol. 89, pp. 890-900, 2016. DOI: 10.1016/j.foodres.2015.10.030

S. Zapata Bustamante, A. Tamayo Tenorio, and B. Alberto Rojano, “ Effect of Roasting on the Secondary Metabolites and Antioxidant Activity of Colombian Cocoa Clones”, Revista Facultad Nacional de Agronomía Medellín, vol. 68, n.o 1, pp. 7497-7507, 2015. DOI: 10.15446/rfnam.v68n1.47836

D. B. Haytowitz y S. Bhagwat, “USDA Database for the Oxygen Radical Absorbance Capacity (ORAC) of Selected Foods, Release 2”, U.S. Department of Agriculture, Beltsville, Maryland, Database, may 2010.

N. Niemenak, C. Rohsius, S. Elwers, D. Omokolo Ndoumou, and R. Lieberei, “Comparative study of different cocoa (Theobroma cacao L.) clones in terms of their phenolics and anthocyanins contents”, Journal of Food Composition and Analysis, vol. 19, n.o 6-7, pp. 612-619, 2006. DOI: 10.1016/j.jfca.2005.02.006

D. A. Jacobo-Velázquez and L. Cisneros-Zevallos, “Correlations of Antioxidant Activity against Phenolic Content Revisited: A New Approach in Data Analysis for Food and Medicinal Plants”, Journal of Food Science, vol. 74, n.o 9, pp. R107-R113, 2009. DOI: 10.1111/j.1750-3841.2009.01352.x

S. A. B. E. van Acker et al., “A Quantum Chemical Explanation of the Antioxidant Activity of Flavonoids”, Chem Res Toxicol, vol. 9, n.o 8, pp. 1305-1312, 1996. DOI: 10.1021/tx9600964

M. Sato, H. Toyazaki, Y. Yoshioka, N. Yokoi, and T. Yamasaki, “Structural Characteristics for Superoxide Anion Radical Scavenging and Productive Activities of Green Tea Polyphenols Including Proanthocyanidin Dimers”, Chem. Pharm. Bull., vol. 58, n.o 1, pp. 98-102, 2010. DOI: 10.1248/cpb.58.98

S. M. Henning, C. Fajardo-Lira, H. W. Lee, A. A. Youssefian, V. L. W. Go, and D. Heber, “Catechin Content of 18 Teas and a Green Tea Extract Supplement Correlates With the Antioxidant Capacity”, Nutrition and Cancer, vol. 45, n.o 2, pp. 226-235, 2003.

L. Gu, S. E. House, X. Wu, B. Ou, and R. L. Prior, “Procyanidin and Catechin Contents and Antioxidant Capacity of Cocoa and Chocolate Products”, J. Agric. Food Chem., vol. 54, n.o 11, pp. 4057-4061, 2006. DOI: 10.1021/jf060360r

B. L. White, L. R. Howard, and R. L. Prior, “Polyphenolic Composition and Antioxidant Capacity of Extruded Cranberry Pomace”, J. Agric. Food Chem., vol. 58, n.o 7, pp. 4037-4042, 2010. DOI: 10.1021/jf902838b.

C. A. Rice-Evans, N. J. Miller, and G. Paganga, “Structure-antioxidant activity relationships of flavonoids and phenolic acids”, Free Radic. Biol. Med., vol. 20, n.o 7, pp. 933-956, 1996. DOI: 10.1016/0891-5849(95)02227-9.

C. A. Rice-Evans, N. Miller, and G. Paganga, “Antioxidant properties of phenolic compounds”, Trends in Plant Science, vol. 2, n.o 4, pp. 152-159,1997. DOI: 10.1016/S1360-1385(97)01018-2

M. S. Fernández-Pachón, D. Villano, A. M. Troncoso, and M. C. García-Parrilla, “Determination of the phenolic composition of sherry and table white wines by liquid chromatography and their relation with antioxidant activity”, Analytica Chimica Acta, vol. 563, n.o 1-2, pp. 101-108, 2006.

L. Stahl et al., “Preservation of Cocoa Antioxidant Activity, Total Polyphenols, Flavan-3-ols, and Procyanidin Content in Foods Prepared with Cocoa Powder”, J. Food Sci., vol. 74, n.o 6, pp. C456-C461, 2009. DOI: 10.1111/j.1750-3841.2009.01226.x

M. Gültekin-Özgüven, I. Berktaş, and B. Özçelik, “Change in stability of procyanidins, antioxidant capacity and in-vitro bioaccessibility during processing of cocoa powder from cocoa beans”, LWT - Food Science and Technology, vol. 72, pp. 559-565, 2016. DOI: 10.1016/j.lwt.2016.04.065.

L. C. Carrillo, J. Londoño-Londoño, and A. Gil, “Comparison of polyphenol, methylxanthines and antioxidant activity in Theobroma cacao beans from different cocoa-growing areas in Colombia”, Food Research International, vol. 60, pp. 273-280, 2014. DOI: 10.1016/j.foodres.2013.06.019

Licencia

Derechos de autor 2020 Revista Colombiana de Química

Creative Commons License

Esta obra está bajo una licencia internacional Creative Commons Atribución 4.0.

Los autores/as conservarán sus derechos de autor y garantizarán a la revista el derecho de primera publicación de su obra, el cuál estará simultáneamente sujeto a la Licencia de reconocimiento de Creative Commons (CC. Atribución 4.0) que permite a terceros compartir la obra siempre que se indique su autor y su primera publicación en esta revista.

Los autores/as podrán adoptar otros acuerdos de licencia no exclusiva de distribución de la versión de la obra publicada (p. ej.: depositarla en un archivo telemático institucional o publicarla en un volumen monográfico) siempre que se indique la publicación inicial en esta revista.

Se permite y recomienda a los autores/as difundir su obra a través de Internet (p. ej.: en archivos telemáticos institucionales o en su página web) antes y durante el proceso de envío, lo cual puede producir intercambios interesantes y aumentar las citas de la obra publicada. (Véase El efecto del acceso abierto).