Ilex Guayusa Loes Research Articles

Kinetic study of solid-liquid extraction of caffeine in Ilex guayusa Loes

Kinetic study of solid-liquid extraction of caffeine in Ilex guayusa Loes

Exploring the Impact of Infusion Parameters and In Vitro Digestion on the Phenolic Profile and Antioxidant Capacity of Guayusa (Ilex guayusa Loes.) Tea Using Liquid Chromatography, Diode Array Detection, and Electrospray Ionization Tandem Mass Spectrometry.

Guayusa tea is derived from the leaves of the Ilex guayusa Loes. plant, which is native to the Amazon rainforest. Beyond its pleasant sensory properties, Guayusa tea is rich in antioxidants, phenolics, and minerals. In this study, the effects of infusion time, temperature, and solvent conditions on the color, antioxidant capacity, total phenolic content, phenolic profile, and antimicrobial activity of Guayusa (Ilex guayusa Loes.) tea were investigated. Guayusa tea samples were prepared using two different solvents, ethanol and water, with 4, 6, and 8-h infusions at 60 and 70 °C. Liquid chromatography, diode array detection, and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) were used to determine a comprehensive profile of phenolic compounds and to detect differences due to infusion conditions. Moreover, after the Guayusa tea infusion with the highest bioactive properties was determined, the effects of in vitro gastrointestinal digestion on the total phenolic content, antioxidant capacity, and phenolic compounds of the Guayusa tea infusion were measured. Phenolic profile analysis identified 29 compounds, among which chlorogenic acid and its derivatives were predominant. The increase in infusion time was correlated with an elevation in total phenolic content. Significant differences were observed between water and ethanol infusions of Guayusa in terms of phenolics and antioxidants. The total amount of phenolic compounds in the samples prepared with both solvents was found to increase after oral intake, depending on the digestion stage; meanwhile, the amounts of flavonoid compounds and di-O-caffeoylquinic acid derivatives decreased during digestion.

Antibacterial and antifungal activity of guayusa (Ilex guayusa Loes.) Extracts obtained by high pressure techniques using polar solvents.

La resistencia a los antimicrobianos (RAM) representa unos de los problemas más importantes de salud mundial. En países en desarrollo, como América Latina, la RAM provoca miles de muertes cada año debido al bajo o limitado acceso a antibióticos y/o cuidado básico de salud. En esta región, la Amazonía alberga una gran biodiversidad que podría ser fuente para el descubrimiento de nuevas substancias antimicrobianas que podrían ayudar a enfrentar la RAM. La guayusa (Ilex guayusa Loes.) ha sido usada durante milenos por los nativos de esta región, y se ha reportado como fuente de compuestos bioactivos. Sin embargo, los compuestos bioactivos obtenidos, han sido mediante la aplicación de técnicas convencionales, pocas investigaciones han aplicado técnicas no convencionales de extracción como fluido supercrítico (ESC) o líquido presurizado (ELP) usando solventes o cosolventes polares. En el presente estudio evaluó la actividad antibacteriana y antifúngica de extractos de guayusa obtenidos mediante técnicas no convencionales ESC y PLE aplicando solventes/cosolventes polares y comparada con la técnica convencional de extracción por maceración. Los solventes/cosolventes usados en las diferentes extracciones fueron: CO2/agua, CO2/etanol/agua, etanol/agua o agua. A partir de la evaluación de la concentración inhibitoria mínima (CIM) de los extractos, se evidenció que presentaron actividad antifúngica contra diferentes dermatofitos: Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes and Trichophyton rubrum en el rango 1000 µg mL-1 a 62.5 µg mL-1. El presente estudio demostró el promisor potencial de los extractos de hojas de guayusa obtenidos mediante baja y alta presión para combatir problemas asociados a dermatofitos.

English

Ilex guayusa Loes. is a medicinal plant used by people of the Ecuadorian Amazon for treating various diseases and for its stimulating properties. In recent years, several studies have shown this plant to possess a high concentration of phenolic compounds and a high antioxidant potential, allowing it to be used as an ingredient for cosmetic formulations. This research confirms the presence of phenolic compounds between 20.18 and 22.44 mg gallic acid equivalents/mL of total phenols and between 7.50 and 3.80 mg hyperoxide equivalents/Ml of total flavonoids, depending on the solvent used (hydroalcoholic or hydroglyceric) to prepare the extract. The evaluation by GC/MS after silylation reveals the presence of 3-hydroxyflavone, techtochrysin, kaempferol and quercetin. The spectrophotometric method of DPPH (2.2-diphenyl-1-picrylhydrazyl) verifies high antioxidant activity in the extracts evaluated and in two cosmetic formulations: Hydrophilic (gel) and lipophilic (cream), with activities similar to ascorbic acid and the natural extract of reference from Camellia sinensis. Clinical evaluation of the lipophilic form shows positive results in increasing skin elasticity and firmness in all volunteers. This study verifies the cosmetic potential of I. guayusa, as well as the abundant concentration of phenolic compounds and its influence on antioxidant activity, with an industrial use in cosmetics, which could become a source of income for the communities where this plant is grown. Key words: Ilex guayusa, natural cosmetics, polyphenols, antioxidants, skin elasticity, skin firmness.

A Novel Technological Breakthrough in Effective Body Recomposition and Weight Management: A Therapeutic Strategy

Obesity and overweight are now defined as an epidemic. An array of evidence substantiates that existing weight loss strategies and programs have not worked. Fat is the lightest of macromolecules, the highest energy reserve of the body, and the last reservoir of survival insurance to be expended. Conventional weight loss concepts and therapies, for the most part, are based on two primary factors; people eat too much and don’t exercise enough. However, with the increase in gym memberships there has also been a significant increase in the purchase of more wholesome foods. The trajectory of obesity therefore should be decreasing; but it’s not. Furthermore, even the culturally ingrained term “weight loss” itself establishes “heaviness” as the primary criteria for evaluating body composition and size, i.e., scale weight, an inaccurate and erroneous measuring criterion owing to additional variables. An additional 10 factors that contribute to a reduction in metabolic rate and an increase in fat storage have been identified. Individually or in combination, these factors are almost totally ignored or omitted from considerations regarding the etiology of obesity. We developed TrimRox, a stimulant‐ and sugar‐free Prodosomed® TRCAP21 nutraceutical technology, utilizing six standardized extracts comprising of D‐ribose nicotinamide complex, Gynostemma pentaphyllum leaf extract, Myrciaria Dubia whole‐berry extract, Ilex guayusa Loes. (Guayusa) leaf extract, Purple Tea Camellia sinensis extract (PurpleForce™), and alpha Glyceryl phosphoryl choline that effectively addresses all the contributing factors. A 21‐day concept validation physician‐supervised pilot study on the TRCAP21 was conducted in 9 subjects to assess changes in various body parts, including chest, upper arms, waist/belly, hips, and thighs, as well as body weight. The results demonstrated that all subjects experienced a significant reduction in size of one or more of these body parts and significant reduction in body weight. A randomized double‐blind placebo‐controlled study is in progress.

High polarity extracts obtained from Ilex guayusa Loes. leaves, exhibit antioxidant capacity

Background: Ilex guayusa Loes. belongs to the family Aquifoliaceae. It is an evergreen tree native to the Amazon region. According to traditional uses, it is used as a diaphoretic, narcotic, purgative, among other uses. Objective: To evaluate the antioxidant capacity, quantify phenols and total flavonoids of extracts obtained from Ilex guayusa leaves. Methods: Total ethanolic extract was obtained using Soxhlet-type equipment, with subsequent liquid/liquid fractionation with solvents of different polarities (petroleum ether, dichloromethane, and ethyl acetate); were reacted with 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+), determining the inhibitory concentration 50 (IC50) and relative antioxidant activity (%AAR), with subsequent statistical analysis (ANOVA and Tukey's post-hoc). Total phenols and flavonoids were quantified using the Folin-Ciocalteu method and complex formation with AlCl3, respectively. Results: The ethanolic extract of leaves showed the highest antioxidant activity in both the DPPH• and ABTS•+ methods with IC50 values of 4.58 and 3.82 ppm, respectively. Conclusions: According to the results obtained in this study, it was possible to corroborate that the ethanolic extract (EE) obtained from the leaves of I. guayusa showed the highest antioxidant capacity, by the DPPH• and ABTS•+ methods. Additionally, it was possible to relate this capacity to the high flavonoids content present in this extract. Based on the above and accompanied by future studies, the Ilex guayusa species could be proposed as an important source of antioxidant compounds with possible application in medicine and the food industry.

Assessing the Genetic Diversity of Ilex guayusa Loes., a Medicinal Plant from the Ecuadorian Amazon

Ilex guayusa Loes. is a shrub native to the Neotropics, traditionally consumed as an infusion. Despite its cultural value and extensive use, genetic research remains scarce. This study examined the genetic and clonal diversity of guayusa in three different Ecuadorian Amazon regions using 17 species-specific SSR markers. The results obtained suggest a moderately low degree of genetic diversity (He = 0.396). Among the 88 samples studied, 71 unique multilocus genotypes (MLGs) were identified, demonstrating a high genotypic diversity. A Discriminant Analysis of Principal Components (DAPC) revealed the existence of two genetic clusters. We propose that a model of isolation-by-environment (IBE) could explain the genetic differentiation between these clusters, with the main variables shaping the population’s genetic structure being temperature seasonality (SD × 100) (Bio 4) and isothermality ×100 (Bio 3). Nonetheless, we cannot dismiss the possibility that human activities could also impact the genetic diversity and distribution of this species. This study gives a first glance at the genetic diversity of I. guayusa in the Ecuadorian Amazon. It could assist in developing successful conservation and breeding programs, which could promote the economic growth of local communities and reinforce the value of ancestral knowledge.

Optimisation of ultrasound-assisted extraction of phenolic antioxidants from Ilex guayusa Loes. leaves using response surface methodology

The present study carried out the optimisation of the total polyphenol content (TPC) extraction assisted by ultrasound in Ilex guayusa leaves applying response surface methodology (RSM). Also, the evaluation of the antioxidant activity of the extract obtained under the optimal extraction conditions was performed. The effect of the variables like, time of sonication, temperature, ethanol/water ratio and solid/liquid relationship and the interactions between them were analysed through the use of a factorial design 2ˆ4. The significant factors were considered for the optimisation, employing a Box-Behnken Design, and the TPC as response variables. It was found that a quadratic model was adequate, with an adjusted R2 value of 0.9367. The optimal conditions proposed, by the response surface model were: an extraction temperature of 60 °C, sonication time of 29.9 min and ethanol/water ratio of 76.8/23.2. The optimised leaves extract of I. guayusa show a TPC of 3.46 (±0.17) g gallic acid equivalents/100 g d.w. Radical scavenger activity of the obtained extract at optimum conditions, was performed through the FRAP and ABTS methods, given as result: 0.080 mmol TROLOX equivalents/100 g d.w. and 40.71 μmol TROLOX equivalents/g d.w., respectively. Due to the present findings, I. guayusa extracts can be proposed as a promising component for functional beverages, cosmetic and pharmaceutical formulation.

Biological activity of extracts from guayusa leaves (Ilex guayusa Loes.) obtained by supercritical CO2 and ethanol as cosolvent

To the best of our knowledge, this is the first study to investigate the biological activities and main constituents of extracts from guayusa leaves (Ilex guayusa Loes.) by using supercritical CO2 as solvent and ethanol as cosolvent at different conditions of pressure and temperature and comparing them with Soxhlet technique. The global yield (X0); total phenolic content (TPC), assessed by Folin–Ciocalteu reagent; in vitro antioxidant capacity, by both ABTS•+ and DPPH methods; antimicrobial activity; and the main phytochemicals, based on gas chromatography-mass spectrometry, of the extracts were tested. X0, TPC, and antioxidant capacity were dependent on technique or solvent used and increased when ethanol was used as cosolvent. Antifungal activity was demonstrated against T. rubrum, T. mentagrophytes, M. gypseum and M. canis. The main phytochemicals were caffeine, squalene and α-amyrin. Therefore, known for its caffeine content, guayusa extracts obtained by supercritical fluid extraction showed important nutraceutical ingredients.

Comparative Composition Analysis of the Dried Leaves of <i>Ilex guayusa</i> (Loes.)

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.

Health Benefits of Bioactive Compounds from the Genus Ilex, a Source of Traditional Caffeinated Beverages.

Tea and coffee are caffeinated beverages commonly consumed around the world in daily life. Tea from Camellia sinensis is widely available and is a good source of caffeine and other bioactive compounds (e.g., polyphenols and carotenoids). Other tea-like beverages, such as those from the genus Ilex, the large-leaved Kudingcha (Ilex latifolia Thunb and Ilex kudingcha C.J. Tseng), Yerba Mate (Ilex paraguariensis A. St.-Hil), Yaupon Holly (Ilex vomitoria), and Guayusa (Ilex guayusa Loes) are also traditional drinks, with lesser overall usage, but have attracted much recent attention and have been subjected to further study. This review summarizes the distribution, composition, and health benefits of caffeinated beverages from the genus Ilex. Plants of this genus mainly contain polyphenols and alkaloids, and show diverse health benefits, which, as well as supporting their further popularization as beverages, may also lead to potential applications in the pharmaceutical or nutraceutical industries.

Bioactive triterpenoids from the caffeine-rich plants guayusa and maté

Unlike all other caffeinated plants, guayusa (Ilex guayusa Loes.) and maté (Ilex paraguariensis A. St. Hill) contain high amounts of pentacyclic triterpenoid acids and alcohols. A phytochemical investigation on these plants revealed a similar triterpenoid profile and a content of ursolic acid (0.7–1%) and amyrin esters (up to 0.5%), quite unusual for dietary plants. The major constituent of the amyrin complex from both plants is α-amyrin palmitate (2a), accompanied by lower amounts of its corresponding palmitoleate (2b) and by the corresponding constitutional isomers from the β-series (3a and 3b, respectively). Ursolic acid (1) was identified as the responsible for the activity of maté and guayusa extracts in the activation of TGR5, a nuclear receptor of relevance for the prevention and management of diabetes and metabolic syndrome because of its involvement in the regulation of energy expenditure and insulin sensitivity.

Antimicrobial Activity of Piper marginatum Jacq and Ilex guayusa Loes on Microorganisms Associated with Periodontal Disease.

Background Chronic periodontitis is a multifactorial infectious disease, where multiple bacteria, such as Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum are implicated. The main purpose of researching natural products is to find substances or compounds with antimicrobial activity. Aim The objective of this work was to determine antimicrobial activity from extracts and obtained fractions from Piper marginatum Jacq and Ilex guayusa Loes on P. gingivalis ATCC 33277, F. nucleatum ATCC 25586, and P. intermedia ATCC 25611. Methods Total ethanol extracts were obtained from both plants. Fractions were obtained from total ethanol extracts with amberlite as a stationary phase employing hexane, acetone, and ethanol-water as solvents. Qualitative and quantitative phytochemical characterization was performed on total ethanol extracts from both plants. Antimicrobial activity from total ethanol extracts and fractions from both plants were evaluated on P. gingivalis ATCC 33277, F. nucleatum ATCC 25586, and P. intermedia ATCC by the well diffusion method with Wilkins–Chalgren agar. Results Piper marginatum Jacq total ethanol extract presented antimicrobial activity against all three bacteria, whereas Ilex guayusa Loes was only efficient against P. gingivalis ATCC 33277 and P. intermedia ATCC 25611, with inhibition halos from 9.3 to 30 mm. Ilex guayusa Loes obtained fractions presented antimicrobial activity against all three microorganisms evaluated, with inhibition halos ranging from 9.7 to 18.7 mm. In regards to Piper marginatum Jacq fractions, inhibition halos were between 8.3 and 19 mm, against all three microorganisms evaluated; only hexane fraction did not present antimicrobial activity against F. nucleatum ATCC 25586. Conclusion Piper marginatum Jacq and Ilex guayusa Loes total ethanol extracts and fractions presented outstanding antimicrobial activity against P. gingivalis ATCC 33277, P. intermedia ATCC 25611, and F. nucleatum ATCC 25586.

Assessing the History of Safe Use of Guayusa

Guayusa (Ilex guayusa Loes.) is a herbal tea that has been consumed for centuries as a traditional food in western Amazon regions where it is now valued as an antioxidant and a stimulant agent. Currently there is intense commercial activity to expand the consumption of guayusa as a healthy energy drink in Europe, where it is classified by law as a novel food. European Union novel food legislation permits traditional foods to be placed on the market if a history of safe use in a non-EU country can be established. However, a scientific assessment of the ‘safe’ use of guayusa is lacking. This study investigates the safety of guayusa consumption, analysing provincial hospital admissions data; national disease register data; national toxicology agency call centre data; and national food safety authority data. In doing so we present a crucial analysis for the novel food premarket risk assessment of dried guayusa leaves, based on the well-established use of this traditional food in Ecuador. Within a three-year period there was one minor adverse effect reported nationally, related to the stimulant properties of guayusa. However, there were no hospital presentations, no product safety notifications and no disease register records of guayusa-related illness. Comprehensive records of unrelated food safety risks demonstrate that Ecuador’s surveillance and reporting system has sufficient rigour to identify risks should they exist. We conclude that there is a history of safe use of guayusa in Ecuador. Establishing safe guayusa consumption is an important milestone for the authorisation of dried guayusa leaves as a novel food with claimed health benefits in Europe. Consequently, this study has significance for international product development of guayusa as an antioxidant energy drink.

Changes in phytochemical composition, bioactivity and in vitro digestibility of guayusa leaves (Ilex guayusa Loes.) in different ripening stages.

Guayusa (Ilex guayusa Loes.) leaves, native of the Ecuadorian Amazon, are popularly used for preparing teas. This study aimed to assess the influence of leaf age on the phenolic compounds and carotenoids and the bioactivity and digestibility (in vitro) of aqueous and hydroalcoholic leaf extracts. In total, 14 phenolic compounds were identified and quantified. Chlorogenic acid and quercetin-3-O-hexose were the main representatives of the hydroxycinnamic acids and flavonols respectively. Seven carotenoids were quantified, lutein being the main compound. Ripening affected phenolic content significantly, but there was no significant difference in carotenoid content. Antioxidant capacity, measured by the DPPH• method, was also significantly affected by leaf age. The measurement of in vitro digestibility showed a decrease in phenolic content (59%) as well as antioxidant capacity, measured by the ABTS•+ method, in comparison with initial conditions of the guayusa infusion. Antibacterial and anti-inflammatory activities were assayed with young leaves owing to their higher phenolic contents. Guayusa did not show any antibacterial activity against Escherichia coli ATCC 25922 or Staphylococcus aureus ATCC 25923. Finally, the hydroalcoholic and aqueous extracts exhibited high in vitro anti-inflammatory activity (>65%). Young guayusa leaves have potential applications as a functional ingredient in food and pharmaceutical industries. © 2017 Society of Chemical Industry.

Guayusa (Ilex guayusa L.) new tea: phenolic and carotenoid composition and antioxidant capacity.

Guayusa (Ilex guayusa Loes) is an evergreen tree native of South America that grows particularly in the upper Amazon region of Ecuador. For its health benefits, it has been cultivated and consumed since ancient times by Amazon indigenous tribes. A total of 14 phenolic compounds were identified and quantified. Chlorogenic acid and quercetin-3-O-hexose were the main representatives of the hydroxycinnamic acids and flavonols, respectively. Five carotenoids were identified, showing lutein the highest concentration. Guayusa leaves revealed high antioxidant capacity determined by two analytical methods, DPPH and ORAC. The industrial processing applied to the leaves modified the composition of bioactive compounds and antioxidant capacity of guayusa. In general, blanched guayusa retained the concentration of phenolic compounds and some carotenoids and similar antioxidant capacity as untreated green leaves. In contrast, fermentation reduced the content of bioactive compounds and showed the lowest antioxidant capacity. Therefore, blanched guayusa has potential for product development as a functional ingredient in the food industry. © 2017 Society of Chemical Industry.

Caracterización de una infusión de cascarilla de cacao (Theobroma cacao L., var. Arriba) con hierbas aromáticas

In this study an assessment through sensory analysis and physico-chemical characterization of prepared infusions made of the cocoa husks, discarded during the roasting process to obtain cocoa paste, mixed with aromatic herbs was performed. Infusion of the husks from Cocoa Arriba (Theobroma cacao L.) was prepared mixed with 3 different varieties of dried herbs, including guayusa (Ilex guayusa Loes), lemongrass (Cymbopogon citratus (DC.) Stapf) and peppermint (Mentha piperita L.) in varying proportions, that sought appropriate considerations of quality and safety specifications demanded by consumers. An experimental design with 2 factors at 3 levels was applied, with varying percentages of husks (a0 : 80%, a1 :70%, a2 : 60%) and dried herbs (b0 : lemongrass, b1 : guayusa, b2 : peppermint), which accounted for a total of 9 treatments plus a control (100% husks). The assays were performed in triplicates. The cocoa husks had 9.78 ± 0.12% moisture, 5.37% lipids, a content of total polyphenolics of 2135 ± 10; 468 ± 10; 662 ± 10 and 977 ± 10 mg GAE (gallic acid equivalent) 100 g-1 sample in the cacao husk, lemongrass, guayusa and peppermint, respectively. A balanced incomplete block design was used for the sensory analysis and treatments were assessed by 18 semi-trained judges, implying five samples per judge, resulting into nine replicates for each treatment performed in duplicate. Sensory analysis included the evaluation of color, odor, flavor and acceptability, and the formulation best valued by the tasters was that containing 80% cocoa husks and 20% guayusa. This formulation had a moisture content of 9.42 ± 0.18%, a pH value of 6.64; soluble solids 0.4 °brix and acidity of 0.014% reported as citric acid, being microbiologically stable at 18 °C with a shelf-life of 148 days.

Actividad antimicrobiana de cuatro plantas del nor-oriente peruano

Se investigó la actividad antimicrobiana in vitro de los extractos metanólicos, etanólicos e hidroalcohólicos de cuatro plantas del nor-oriente peruano: Cassia reticulata (planta entera), Ilex guayusa Loes (hojas), Piper lineatum (hojas), y Terminalia catappa (hojas). Las especies fueron recolectadas en el departamento de Cajamarca, excepto Terminalia catappa (Amazonas). La actividad antimicrobiana se evaluó mediante el método de difusión en agar. Los microorganismos utilizados fueron las bacterias Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Bacillus subtilis y Escherichia coli; y los hongos Candida albicans, Aspergillus niger y Microsporum canis. De doce extractos investigados, ocho (67%) presentaron actividad antimicrobiana significativa frente a Staphylococcus aureus y Staphylococcus epidermidis y uno (8%) frente a Escherichia coli. De doce extractos investigados, diez (83%) presentaron actividad significativa frente a Candida albicans, y seis (50%) contra Microsporum canis. Los extractos con la mejor actividad actimicrobiana fueron los tres extractos del Piper lineatum; el extracto hidroalcohólico de Cassia reticulata y el hidroalcohólico de Terminalia catappa.