Journal of Food and Nutrition Research
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Journal of Food and Nutrition Research. 2018, 6(10), 638-644
DOI: 10.12691/jfnr-6-10-4
Open AccessArticle

Comparative Composition Analysis of the Dried Leaves of Ilex guayusa (Loes.)

Graham Wise1, 2, and Demetrio E. Santander3

1Office of the Vice President of Innovation, Universidad Regional Amazónica Ikiam, Tena, Ecuador, 150150

2Center for Global Health Equity, University of Wisconsin-Milwaukee, Milwaukee, USA, 53201

3Waykana Guayusa, Quito, Ecuador, 170150

Pub. Date: November 09, 2018

Cite this paper:
Graham Wise and Demetrio E. Santander. Comparative Composition Analysis of the Dried Leaves of Ilex guayusa (Loes.). Journal of Food and Nutrition Research. 2018; 6(10):638-644. doi: 10.12691/jfnr-6-10-4


Guayusa (Ilex guayusa Loes.) is a traditional herbal tea of western Amazon regions and an international commodity of increasing importance. Its consumption is rapidly growing in the USA and Canada, and the authorization of guayusa extract as a novel food in Europe signals further international market growth of this antioxidant and stimulant tea. However, little is known about the chemical composition of guayusa, despite much research on related Ilex species. There is an urgent need for a deeper understanding of the chemical composition of guayusa, to support assessments of its safety and its claimed nutritional value. This study follows the novel food assessment framework of the European Union Food Safety Authority, characterizing the proximate composition of guayusa and elucidating caffeine, amino acid and elemental components. It also evaluates potential microbial, mycotoxin and pesticide residue contaminants. The chemical composition of guayusa is analyzed in context with the related and well-characterized foods, yerba mate (Ilex paraguariensis A. St.-Hil.) and tea (Camellia sinensis L.). Guayusa’s moisture content, caffeine concentration, amino acid compliment and elemental profile, including heavy metals, present no greater risk to human health than the consumption of tea or yerba mate. It is also established that there is a low risk of mycotoxin, bacterial or pesticide residue contamination of guayusa. The chemical composition of guayusa presents no barrier to the authorization of guayusa as a novel food in accordance with European Union novel food guidelines.

guayusa yerba mate traditional food novel food antioxidant stimulant herbal tea

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[1]  Galle, F.C., Hollies. The Genus Ilex, Timber Press, Portland, 1997, 573.
[2]  Krause, T. and Ness, B., “Energizing agroforestry: Ilex guayusa as an additional commodity to diversify amazonian agroforestry systems”, International Journal of Biodiversity Science, Ecosystem Services and Management, 13 (1). 191-203. 2017.
[3]  Innerhofer, S. and Bernhardt, K.G., “Ethnobotanic garden design in the Ecuadorian Amazon”, Biodiversity and Conservation, 20 (2). 429-439. 2011.
[4]  Dueñas, J.F., Jarrett, C., Cummins, I. and Logan–Hines, E., “Amazonian Guayusa (Ilex guayusa Loes.): A Historical and Ethnobotanical Overview”, Economic Botany, 70 (1). 85-91. 2016.
[5]  Schultes, R.E., “Discovery of an ancient guayusa plantation in Colombia”, Botanical Museum Leaflets, 27 (5-6). 143-153. 1979.
[6]  Bennett, B.C., “Hallucinogenic plants of the Shuar and related indigenous groups in Amazonian Ecuador and Peru”, Brittonia, 44 (4). 483-493. 1992.
[7]  Bussmann, R.W., Malca-García, G., Glenn, A., Sharon, D., Chait, G., Díaz, D., Pourmand, K., Jonat, B., Somogy, S., Guardado, G., Aguirre, C., Chan, R., Meyer, K., Kuhlman, A., Townesmith, A., Effio-Carbajal, J., Frías-Fernandez, F. and Benito, M., “Minimum inhibitory concentrations of medicinal plants used in Northern Peru as antibacterial remedies”, Journal of Ethnopharmacology, 132 (1), 101-108. 2010.
[8]  Contero, F., Abdo, S., Vinueza, D., Moreno, J., Tuquinga, M., Paca, N., “Estrogenic activity of ethanolic extracts from leaves of Ilex guayusa Loes. and Medicago sativa in Rattus norvegicus”, Pharmacologyonline, 2. 95-99. 2015.
[9]  Sidali, K.L., Morocho, P.Y. and Garrido-Pérez, E.I., “Food tourism in indigenous settings as a strategy of sustainable development: The case of Ilex guayusa Loes. in the Ecuadorian Amazon”, Sustainability (Switzerland), 8 (10). 967. 2016.
[10]  Wise, G. and Santander, D.E., “Assessing the history of safe use of guayusa”, Journal of Food and Nutrition Research, 6 (7). 471-475. 2018.
[11]  Commission Implementing Regulation (EU) 2017/2470 of 20 December 2017 establishing the Union list of novel foods in accordance with Regulation (EU) 2015/2283 of the European Parliament and of the Council on novel foods, (EU) 2017/2470 C.F.R. § L 351. 2017.
[12]  Advisory Committee on Novel Foods, Record of views formed in response to inquiries, FSANZ, Canberra, 2018.
[13]  Knudsen, I., Søborg, I., Eriksen, F.D., Pilegaard, K. and Pedersen, J.W., Risk Assessment and Risk Management of Novel Plant Foods: Concepts and Principles. Nordic Council of Ministers, Copenhagen, 2005, 108.
[14]  Turck, D., Bresson, J-L., Burlingame, B., Dean, T., Fairweather-Tait, S., Heinonen, M., Hirsch-Ernst, K. I., Mangelsdorf, I., McArdle, H., Naska, A., Neuhauser-Berthold, M., Nowicka, G., Pentieva, K., Sanz, Y., Siani, A., Sjodin, A., Stern, M., Tome, D., Vinceti, M., Willatts, P., Engel, K-H., Marchelli, R., Poting, A., Poulsen, M., Schlatter, J., Gelbmann, W., de Sesmaisons-Lecarre, A., Verhagen, H. and van Loveren, H., “Guidance on the preparation and presentation of thenotification and application for authorisation of traditional foods from third countries in the context of Regulation (EU) 2015/2283”, EFSA Journal, 14 (11). 16. 2016.
[15]  Howlett, J., Edwards, D.G., Cockburn, A., Hepburn, P., Kleiner, J., Knorr, D., Kozianowski, G., Müller, D., Peijnenburg, A., Perrin, I., Poulsen, M. and Walker, R., The safety assessment of Novel Foods and concepts to determine their safety in use. Journal of Food Science and Nutrition, 54 (Supp1), S1-32. 2003.
[16]  Yi, F., Zhao, X-L., Peng, Y. and Xiao, P-G, “Genus Ilex L.: phytochemistry, ethnopharmacology, and pharmacology”, Chinese Herbal Medicines, 8 (3). 209-230. 2016.
[17]  Linck, V., de Sá, I.M. and Elisabetsky, E., “Yerba mate or Paraguay tea”, Chinese Herbal Medicines, 6. 253-255. 2014.
[18]  Graham, H.N., “Tea: the plant and its manufacture; chemistry and consumption of the beverage”, Progress in Clinical and Biological Research, 158. 29-74. 1984.
[19]  Jara, A., Rodriguez, Y., Cornejo, J., Cazar, ME., Gutierrez, M. and Astudillo, L. “Antioxidant activity and total phenolics of plants used in traditional medicine in Ecuador”, in The 17th International Electronic Conference on Synthetic Organic Chemistry, Sciforum Electronic Conference Series, b001. 2013.
[20]  García-Ruiz, A., Baenas, N., Benítez-González, A.M., Stinco, C.M., Meléndez-Martínez, A.J., Moreno, D.A., and Ruales, J., “Guayusa (Ilex guayusa L.) new tea: phenolic and carotenoid composition and antioxidant capacity”, Journal of the Science of Food and Agriculture, 97 (12). 3929-3936. 2017.
[21]  García-Ruiz, A., Baenas, N., Benítez-González, A.M., Stinco, C.M., Meléndez-Martínez, A.J., Moreno, D.A. and Ruales, J., “Bioactive compounds and antioxidant capacity of green and processed leaves of guayusa (Ilex paraguariensis Loes.)”, in Proceedings of the II International Conference on Food Chemistry & Technology (FCT-2016), Journal of Food Chemistry & Nanotechnology, 2 (Suppl. 4). S8-S9. 2016.
[22]  Villacís-Chiriboga, J., García-Ruiz, A., Baenas, N., Moreno, D.A., Meléndez-Martínez, A.J., Stinco, C. M., Jerves-Andrade, L., León-Tamariz, F., Ortiz-Ulloa, J. and Ruales, J., “Changes in phytochemical composition, bioactivity and in vitro digestibility of guayusa leaves (Ilex guayusa Loes.) in different ripening stages”. Journal of the Science of Food and Agriculture, 98 (5). 1927-1934. 2018.
[23]  Pardau, M.D., Pereira, A.S.P., Apostolides, Z., Serema, J.C. and Bester, M.J., “Antioxidant and anti-inflammatory properties of Ilex guayusa tea preparations: a comparison to Camellia sinensis teas”, Food and Function, 8 (12). 4601-4610. 2017.
[24]  Moldoveanu, S.C. and Scott, W.A., “Analysis of four pentacyclic triterpenoid acids in several bioactive botanicals with gas and liquid chromatography and mass spectrometry detection”, Journal of Separation Science, 39 (2). 324-332. 2016.
[25]  Moldoveanu, S., Scott, W. and Zhu, J. “Analysis of small carbohydrates in several bioactive botanicals by gas chromatography with mass spectrometry and liquid chromatography with tandem mass spectrometry”, Journal of Separation Science, 38 (21). 3677-3686. 2015.
[26]  Moldoveanu, S.C., Zhu, J. and Qian, N., “Free amino acids analysis by liquid chromatography with tandem mass spectrometry in several botanicals with antioxidant character”, Journal of Separation Science, 38 (13). 2208-2221. 2015.
[27]  Lewis, W.H., Elvin-Lewis, M.P.F., and Gnerre, J.C., Introduction to the ethnobotanical pharmacopeia of the Amazonian Jivaro of Peru, in Medicinal and Poisonous Plants of the Tropics, Pudoc, Wageningen, 1987, 102.
[28]  Lewis, W.H., Kennelly, E.J., Bass, G.N., Wedner, H.J., Elvin-Lewis, M.P. and D. Fast, W. “Ritualistic use of the holly Ilex guayusa by Amazonian Jivaro Indians”, Journal of Ethnopharmacology, 33 (1-2). 25-30. 1991.
[29]  Radice, M. and Vidari, G., “Caracterización fitoquímica de la especie Ilex guayusa Loes. y elaboración de un prototipo de fitofármaco de interés comercial”, La Granja, 6. 3-10. 2000.
[30]  Barriga-Coronel, G., Determinacion del Contenido de Cafeína en Infusiones de hoya de guayusa (Ilex guayusa) bajo dos condiciones de secado, Universidad de los Andes, Quito, 2017.
[31]  Cobos Morales, L.A., Determinación del Contenido de Cafeína en un Cultivo Comercial de Guayusa (Ilex guayusa), Universidad Central del Ecuador, Quito, 2017.
[32]  Melo Gallegos, V.A., Composición y Análisis Químico de la Especie Ilex guayusa Loes., Universidad San Francisco de Quito, Quito, 2014.
[33]  Holmstedt, B. and Lindgren, J.-E., “Alkaloid analyses of botanical material more than a thousand years old”, in A Medicine Man's Implements and Plants in a Tiahuanacoid Tomb in Highland Bolivia, S.H. Wassen, ed., Goteborgs Ethnografiska Museum, Goteborg. 1972. 139-144.
[34]  McNeely, J.A. and Schroth, G., “Agroforestry and biodiversity bonservation – traditional practices, present dynamics, and lessons for the future”, Biodiversity & Conservation, 15 (2). 549-554. 2006.
[35]  Jose, S., “Agroforestry for conserving and enhancing biodiversity”, Agroforestry Systems, 85 (1). 1-8. 2012.
[36]  Jose, S., “Agroforestry for ecosystem services and environmental benefits: an overview”, Agroforestry Systems, 76 (1). 1-10. 2009.
[37]  AOAC International, Official methods of analysis of AOAC International, 20th Ed., AOAC International, Rockville, 2016.
[38]  American Oil Chemists' Society, Official Methods and Recommended Practices of the AOCS. 7th Ed., AOCS, Urbana, 2017.
[39]  Food and Agriculture Organization of the United Nations, Guidelines on the Performance Criteria for Methods of Analysis for the Determination of Pesticide Residues in Food and Feed, FAO, Rome, 2017.
[40]  Greenfield, H. and Southgate, D.A.T., Food composition data: Production, management and use. 2nd Ed., Food and Agriculture Organization of the United Nations, Rome, 2003.
[41]  Mohammed, M.I. and Sulaiman, M.A., “Proximate, caffeine and tannin analyses in some brands of tea consumed in Kano metropolis, Nigeria”, Bayero Journal of Pure and Applied Sciences, 2 (2). 19-21. 2009.
[42]  Berté, K.A.S., Beux, M.R., Spada, P.K.W.D.S., Salvador, M. and Hoffmann-Ribani, R., “Chemical composition and antioxidant activity of Yerba-Mate (Ilex paraguariensis A.St.-Hil., Aquifoliaceae) extract as obtained by spray drying”, Journal of Agricultural and Food Chemistry, 59. 5523-5527. 2011.
[43]  Scientific Committee for Food, “Opinion on the potential micro-biological risk arising from the presence of moisture in tea (expressed on 19th September 1997)”, in Reports of the Scientific Committee for Food (44th series), Directorate-Gencral for Consumer Policy and Consumer Health Protection, Belgium, 1998, 78.
[44]  Werkhoven, J., “Tea processing”, in FAO Agricultural Services Bulletin, Food and Agricultural Organization of the United Nations: Rome, 1974.
[45]  Ozdemir, F., Gokalp, H.Y. and Nas, S., “Effects of shooting period, times within shooting periods and processing systems on the extract, caffeine and crude fiber contents of black tea”, Zeitschrift fur Lebensmittel Untersuchung und Forschung, 197 358-362. 1993.
[46]  Chen, L., Zhou, Z.-X. and Yang, Y.-J., “Genetic improvement and breeding of tea plant (Camellia sinensis) in China: from individual selection to hybridization and molecular breeding”, Euphytica, 154 (1-2). 239-248. 2007.
[47]  Villacís-Chiriboga, J., “Etnobotánica y systemas tradicionalies de salud en Ecuador, Enfoque en la guayusa (Ilex guayusa)”, Revista Etnobiologia, 15 (3). 79-88. 2017.
[48]  Heck, C.I. and Mejida, E.G., “Yerba mate tea (Ilex paraguariensis): A comprehensive review on chemistry, health implications, and technological considerations”, Journal of Food Science, 72 (9). R138-R162. 2007.
[49]  Perva-Uzunalic, A., Skerget, M., Knez, Z., Weinreich, B., Otto, F. and Gruner, S., “Extraction of active ingredients from green tea (Camellia sinensis): Extraction efficiency of major catechins and caffeine”, Food Chemistry, 96. 597-606. 2006.
[50]  Vázquez, M. and Vélez, D., “Other foods of plant origin”, in Handbook of Mineral Elements in Food, M. de la Guardia and S. Garrigues, Eds, Wiley Blackwell, West Sussex, 2015, 699-708.
[51]  Giulian, R., et al., Iochims dos Santos, C.E., de Moraes Shubeita, S., da Silva, L.M., Dias, J.F. and Yoneama, M.L., “Elemental characterization of commercial mate tea leaves (Ilex paraguariensis A. St.-Hil.) before and after hot water infusion using ion beam techniques”, Journal of Agricultural and Food Chemistry, 55. 741-746. 2007.
[52]  Marcelo, M.C.A., Martins, C.A., Pozebon, D., Dressler, V.L. and Ferrão, M.F., “Classification of yerba mate (Ilex paraguariensis) according to the country of origin based on element concentrations”, Microchemical Journal, 117. 164-171. 2014.
[53]  Froes, R.E.S., et al., Neto, V.B., Beinner, M.A., Nascentes, C.C. and da Silva, J.B.B., “Determination of inorganic elements in teas using inductively coupled plasma optical emission spectrometry and classification with exploratory analysis”, Food Analytical Methods, 7. 540-546. 2014.
[54]  da Costa, A.M.G., et al., Nogami, E.M., Visentainer, J. V., de Souza, N.E. and Garcia, E.E., “Fractionation of aluminum in commercial green and roasted yerba mate samples (Ilex paraguariensis St. Hil.) and in their infusions”, Journal of Agricultural and Food Chemistry, 57 (1). 196-200. 2009.
[55]  Flaten, T.P., “Aluminium in tea-concentrations, speciation and bioavailability”, Coordination Chemistry Reviews, 228 (2). 385-395. 2002.
[56]  Shen, F.M. and Chen, H.W., “Element composition of tea leaves and tea infusions and its impact on health”, Bulletin of Environmental Contamination and Toxicology, 80. 300-304. 2008.
[57]  Szymczycha-Madeja, A., Welna, M. and Pohl, P., “Elemental analysis of teas and their infusions by spectrometric methods”. Trends in Analytical Chemistry, 35. 165-181. 2012.
[58]  Wang, J., Cheung, W. and Leung, D., “Determination of Pesticide Residue Transfer Rates (Percent) from Dried Tea Leaves to Brewed Tea”, Journal of Agricultural and Food Chemistry, 62 (4). 966-983. 2014.
[59]  Hossain, M.M., Karim, R., Begum, S., Islam, M.G.R. and Hoque, M.M., “Assessment of microbial load in made tea and antimicrobial property of made tea infusion”, International Journal of Public Health Research, 3 (2). 276-281. 2013.
[60]  Institute of Medicine and National Research Council, Scientific Criteria to Ensure Safe Food. National Academies Press, Washington, D.C., 2003.
[61]  Tea and Herbal Infusions Europe, Compendium of Guidelines for Tea (Camellia Sinensis), Tea and Herbal Infusions Europe, Hamberg, 2014, 43.
[62]  Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs, (EU) 1881/2006 C.F.R. § L 364 (2006).