Acute Effects of Naturally Occurring Guayusa Tea and Nordic Lion's Mane Extracts on Cognitive Performance.

The aim of this study was to assess the effects of guayusa extract and Nordic Lion's Mane (LM) on cognition. Using a randomized, double-blind, placebo-controlled, crossover design, we examined the effects of a single dose of 650 mg guayusa extract (AMT: AmaTea® Max) vs. 1 g Nordic-grown Lion's Mane (LM) vs. placebo (PL). Participants attended three testing visits consisting of neuropsychological tests (Go/No-go, N-Back, and Serial 7 s tasks) assessing performance, subjective assessments of cognitive perception, and vital signs. Each assessment was measured at baseline (pre-ingestion) and 1 and 2 h post ingestion. AMT significantly (p ≤ 0.05) improved the number of attempts during Serial 7s, total score, number of correct responses, total number of responses, and reaction time during N-Back and improved Go stimulus reaction time, but it reduced the percentage of correct responses in the No-go stimulus response during Go/No-go. LM significantly (p ≤ 0.05) improved the number of attempts during Serial 7s and reaction time during N-Back and improved Go stimulus reaction time in Go/No-go. AMT improved mental clarity, focus, concentration, mood, and productivity at 1 and 2 h (p < 0.05); the ability to tolerate stress at 1 h; and had greater ratings than LM and PL for mental clarity, focus, concentration, and productivity. PL improved focus and concentration at 1 h from baseline (p ≤ 0.05). AMT and LM improved subjective ratings of "happiness compared to peers" and "getting the most out of everything" (p < 0.05); however, this occurred earlier in LM (i.e., 1 h post ingestion). AMT uniquely elevated blood pressure from baseline. AMT significantly improved cognitive performance and self-perceived cognitive indices of affect over a 2 h period and perceptions of happiness 2 h post ingestion. In comparison, LM helped improve working memory, complex attention, and reaction time 2 h post ingestion and perceptions of happiness over a 2 h period.

High polarity extracts obtained from Ilex guayusa Loes. leaves, exhibit antioxidant capacity / Extractos de alta polaridad obtenidos a partir de las hojas de Ilex guayusa Loes. exhiben capacidad antioxidante

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

Antecedentes:Ilex guayusa Loes. pertenece a la familia Aquifoliaceae. Es un árbol perenne, nativo de la región amazónica. De acuerdo con los usos tradicionales es empleada como diaforético, narcótico, purgativo, entre otros usos. Objetivo: Evaluar la capacidad antioxidante, cuantificar fenoles y flavonoides totales de extractos obtenidos a partir de las hojas de Ilex guayusa. Métodos: El extracto etanólico total se obtuvo mediante un equipo tipo "Soxhlet", con posterior fraccionamiento líquido/líquido con solventes de diferentes polaridades (éter de petróleo, diclorometano y acetato de etilo); se hicieron reaccionar con 2,2-difenil-1-picrilhidracilo (DPPH•) y ácido 2,2'-azino-bis-3-etilbenzotiazolin-6-sulfónico (ABTS•+), determinando la concentración inhibitoria 50 (IC50) y la actividad antioxidante relativa (%AAR), con posterior análisis estadístico (ANOVA y post-hoc de Tukey). Los fenoles y flavonoides totales se cuantificaron con el método de Folin-Ciocalteu y el de formación de complejos con AlCl3, respectivamente. Resultados: El extracto etanólico de las hojas fue el que presentó mayor actividad antioxidante tanto en el método DPPH• como en el ABTS•+ con valores de IC50 de 4,58 y 3,82 ppm, respectivamente. Conclusiones: De acuerdo con los resultados obtenidos en este estudio, se pudo corroborar que el extracto etanólico obtenido de las hojas de I. guayusamostró la mayor capacidad antioxidante, por los métodos DPPH• y ABTS•+. Adicionalmente, se pudo relacionar esta capacidad con el alto contenido de flavonoides presentes en este extracto. Con base en lo anterior y acompañado de futuros estudios, la especie I. guayusa podría proponerse como una importante fuente de compuestos antioxidantes con posible aplicación en los campos de la medicina y la industria alimentaria

Effect of solvent-solvent partition on antioxidant activity and GC-MS profile of Ilex guayusa Loes. leaves extract and fractions.

The use of guayusa (Ilex guayusa Loes.) leaves as functional food has increase recently. This work discusses the antioxidant activity and volatile compounds of guayusa leaves extract and fractions. The methanol crude extract was obtained by maceration, subsequently hexane, chloroform, ethyl acetate, and aqueous fractions were collected by solvent-solvent partition. Total phenolic content (TPC), total flavonol/flavone content (TFC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and ferric reducing antioxidant power (FRAP) were measured by ultraviolet-visible (UV-Vis) spectrophotometry. The results revealed that ethyl acetate fraction showed highest inhibition against DPPH radical (93.86 ± 0.95%) at 500 µg/mL, and reduce the ferric-tripyridyltriazine complex (Fe3+-TPTZ) at 1619.81 mg trolox equivalent (TE)/g, followed by aqueous fraction. This bioactivity could be related to phenolic acids, flavones and flavonols content, as well as the caffeine, dodecanoic acid isopropyl ester, caffeic acid, and malic acid identified by gas chromatography-mass spectrometry (GC-MS). These findings support the antioxidant properties of this plant material.

[Triterpenoids from leaves of Ilex guayusa].

The chemical constituents from the leaves of Ilex guayusa were investigated. Sixteen triterpenoids were isolated from the 95% ethanol extract of dried leaves of I. guayusa by silica gel, Sephadex LH-20, and ODS column chromatographies and semi-prepa-rative HPLC. Those triterpenoids were identified by NMR, HR-MS, and literature analysis: 3ß-hydroxy-11α,12α-epoxy-24-nor-urs-4(23)-ene-28,13ß-olide(1), 3ß-hydroxy-24-nor-4(23),12-oleanadien-28-methyl ester(2), oleanolic acid(3), 3ß,28-dihydroxy-12-oleanene(4), 2α,3ß-dihydroxy-11α,12α-epoxy-24-'nor-olean-4(23)-ene-28,13ß-olide(5), ursolic acid(6), 3ß,23-dihydroxy ursolic acid(7), 3ß,28-dihydroxy-12-ursene(8), 3ß-28-nor-urs-12-ene-3,17-diol(9), 3ß-hydroxyurs-11-ene-28,13ß-olide(10), 13ß,28-epoxy-3ß-hydroxy-11-ursene(11), 3ß-hydroxy-28,28-dimethoxy-12-ursene(12), 3ß-hydroxy-24-nor-urs-4(23),12-dien-28-oic acid(13), 3ß-hydroxy-24-nor-urs-4(23),12-dien-28-methyl ester(14), 2α,3ß-dihydroxy-11α,12α-epoxy-24-nor-urs-4(23)-ene-28,13ß-olide(15) and 2α,3ß-dihydroxy-11α,12α-epoxy-24-nor-urs-4(23),20(30)-dien-28,13ß-olide(16). Compounds 1-2 were new compounds, and compounds 4-5, 7 and 9-16 were isolated from I. guayusa for the first time.

Triterpenoids from leaves of Ilex guayusa / 中国中药杂志

The chemical constituents from the leaves of Ilex guayusa were investigated. Sixteen triterpenoids were isolated from the 95% ethanol extract of dried leaves of I. guayusa by silica gel, Sephadex LH-20, and ODS column chromatographies and semi-prepa-rative HPLC. Those triterpenoids were identified by NMR, HR-MS, and literature analysis: 3β-hydroxy-11α,12α-epoxy-24-nor-urs-4(23)-ene-28,13β-olide(1), 3β-hydroxy-24-nor-4(23),12-oleanadien-28-methyl ester(2), oleanolic acid(3), 3β,28-dihydroxy-12-oleanene(4), 2α,3β-dihydroxy-11α,12α-epoxy-24-'nor-olean-4(23)-ene-28,13β-olide(5), ursolic acid(6), 3β,23-dihydroxy ursolic acid(7), 3β,28-dihydroxy-12-ursene(8), 3β-28-nor-urs-12-ene-3,17-diol(9), 3β-hydroxyurs-11-ene-28,13β-olide(10), 13β,28-epoxy-3β-hydroxy-11-ursene(11), 3β-hydroxy-28,28-dimethoxy-12-ursene(12), 3β-hydroxy-24-nor-urs-4(23),12-dien-28-oic acid(13), 3β-hydroxy-24-nor-urs-4(23),12-dien-28-methyl ester(14), 2α,3β-dihydroxy-11α,12α-epoxy-24-nor-urs-4(23)-ene-28,13β-olide(15) and 2α,3β-dihydroxy-11α,12α-epoxy-24-nor-urs-4(23),20(30)-dien-28,13β-olide(16). Compounds 1-2 were new compounds, and compounds 4-5, 7 and 9-16 were isolated from I. guayusa for the first time.

A critical review of the composition and history of safe use of guayusa: a stimulant and antioxidant novel food.

Due to rapid international market development, there is a strong and urgent need to assess the safety of the novel food, Ilex guayusa. Guayusa has been consumed for centuries in the western Amazon as an herbal tea, and novel food regulation provide a detailed framework for safety assessment of novel foods with such a history of use. This study reviews guayusa's taxonomy, chemical composition, toxicology, ethnobotany, and history of safe use as key elements of a robust novel food safety assessment. Guayusa is a product of traditional agricultural systems with a continuous history of consumption in Ecuador. Its known chemical composition appears to present no greater risk to human health than existing teas such as green tea or yerba mate, although our understanding of guayusa's chemical profile is still nascent, requiring further investigation. Broad consumption of guayusa is not associated with a history of adverse effects or product safety notifications. Biochemical and phytochemical studies have profiled its nutritional content, metabolite composition, and bioactivity, validating guayusa's antioxidant and stimulant properties. In conclusion, guayusa leaves have a well-documented chemical composition and history of safe use, which are key considerations for authorization as a novel food in the EU.

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 ß-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.

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

BACKGROUND: 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. RESULTS: 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%). CONCLUSION: Young guayusa leaves have potential applications as a functional ingredient in food and pharmaceutical industries. © 2017 Society of Chemical Industry.

Antioxidant and anti-inflammatory properties of Ilex guayusa tea preparations: a comparison to Camellia sinensis teas.

Ilex guayusa tea preparations are now commercially available as Runa tea. Little is known regarding the antioxidant and anti-inflammatory bioactivities of this tea. The I. guayusa teas had a total polyphenolic content between 54.39 and 67.23 mg GAE per g dry mass and peroxyl radical scavenging capacities between 1773.41 and 2019 µmol TE per g dry mass, nearly half of that for the Camellia sinensis teas. The I. guayusa teas afforded 60-80% protection from oxidative stress in the Caco-2 cellular antioxidant assay, comparable to the C. sinensis teas. The anti-inflammatory activity in lipopolysaccharide-stimulated RAW 264.7 cells of I. guayusa teas was similarly comparable to the C. sinensis teas with nitric oxide production reduced by 10-30%. Major compounds identified by mass spectrometry were the phenolic mono- and dicaffeoylquinic acid derivatives. I. guayusa teas are a good source of dietary phenolic compounds with cellular antioxidant and anti-inflammatory properties.

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

BACKGROUND: 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. RESULTS: 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. CONCLUSION: Therefore, blanched guayusa has potential for product development as a functional ingredient in the food industry. © 2017 Society of Chemical Industry.

Etnobotánica y Sistemas Tradicionales de Salud en Ecuador. Enfoque en la Guayusa (Ilex Guayusa Loes) / Ethnobotany and Traditional Health Systems in Ecuador. Focus on the Guayusa (Ilex Guayusa Loes)

El objetivo del presente manuscrito fue exponer la importancia que la etnomedicina posee como fundamento para la identidad cultural y el descubrimiento de nuevos fármacos, enfocado en la Amazonía ecuatoriana y el uso de guayusa (Ilex guayusa Loes.). Desde el inicio de la historia, los seres humanos han hecho uso de los elementos naturales para beneficio propio, principalmente en el área de salud y alimentación. Las plantas son un recurso que ha jugado un papel sustancial en el desarrollo de las civilizaciones por la importancia que tienen dentro de la identidad cultural de los distintos asentamientos humanos, siendo la etnobotánica y la etnomedicina dos disciplinas que a nivel de Latinoamérica se encuentran muy fundamentadas por la tradición que caracteriza a los pueblos aborígenes. En Ecuador, el uso de plantas con fines médicos y tradicionales se ha documentado ampliamente, determinándose que su principal aplicación es como tratamiento para enfermedades "sobrenaturales". En la Amazonía ecuatoriana, la infusión de guayusa (Ilex guayusa Loes.) es consumida en diversas etnias, pues se afirma que tiene efectos estimulantes y provee diferentes beneficios para la salud. Sin embargo, su existencia y por ende las prácticas culturales que se desarrollan en torno al consumo de guayusa, se ven amenazados por el cambio climático y los efectos devastadores que se evidencian en los bosques ecuatorianos. En conclusión, la guayusa tiene una importancia vital en la medicina y cultura en las culturas indígenas amazónicas de Ecuador; así mismo, el estudio científico de esta planta no solo cimentará las bases para el desarrollo de fármacos y compuestos similares, sino que permitirá entender la razón de su uso tan prolongado en el tiempo, y con esto promover su uso y conservación.

The use of medicinal plants by rural populations of the Pastaza province in the Ecuadorian Amazon / Uso de plantas medicinais por populações rurais da província de Pastaza, na Amazônia equatoriana

Mera, Santa Clara and Pastaza municipalities are located in the Ecuadorian Amazon region. The objective of the study was to identify plant species used in traditional medicine by small farmers of these localities, and to classify these plants according to locality, farmer ethnicity and purposes of use. It was also investigated whether the use of medicinal plants differs between the ethnic groups. Data were collected by applying a questionnaire and personal interview with 213 farmers belonging to two ethnicities (Kichwa and mestizo), and to different municipalities (Mera, Santa Clara and Pastaza). Generated data were analyzed using contingency tables and frequency and the most representative species were determined by proportion analysis comparison. A total of 34 families and 52 species of medicinal plants were identified. The most used species was Ilex guayusa which was cited 48 times. Santa Clara municipality and Kichwa farmers used the highest number of species. These species belonged to the Lamiaceae and Solanacease family, and the plants were used for treating stomach pain, cold and inflammations. There were significant differences (Chi square test p < 0.05) between localities and ethnicities (Kichwa and mestizo). There were differences in the use of medicinal plant species among members of the Kichwa ethnicity and mestizo farmers, depending on locality, being Ilex guayusa the most used species.

A pesquisa foi desenvolvida em três municípios da Província de Pastaza, (Mera, Santa Clara e Pastaza), na Amazônia equatoriana. O objetivo do estudo foi identificar espécies vegetais utilizadas na medicina tradicional pelos agricultores nestas localidades e classificar as espécies segundo a localidade, etnia do produtor e as aflições nas que eram utilizadas. Alem disso, na pesquisa analisaram-se as diferença de uso das plantas entre as etnias Kichwa e Mestiça. A metodologia do trabalho consistiu na aplicação de questionários e entrevistas pessoais com 213 agricultores das diferentes etnias. Utilizaram-se as tabelas de contingência por freqüência de uso com os dados gerados, para determinar as espécies mais representativas e em cada grupo realizou-se comparação por análise de proporções. Os principais resultados mostraram a existência de 52 espécies de plantas medicinais pertencente a 34 famílias. A espécie mais utilizada foi Ilex guayusa com 48 registros. Os produtores Kichwa do município Santa Clara registraram o maior número de espécies pertencente às famílias Lamiaceae e Solanacease e as plantas foram utilizadas para tratamento de dor de estômago, gripe e inflamações. A prova de "chi quadrado" mostrou diferenças (p < 0,05) entre os municípios e as etnias. Conclui-se que existe diferença no uso de plantas medicinais entre as localidades e os grupos étnicos estudados. A espécie Ilex guayusa foi a planta medicinal mais usada pelos agricultores independentemente da localidade e a etnia.

General and Genetic Toxicology of Guayusa Concentrate (Ilex guayusa).

Tea from the leaves of guayusa (Ilex guayusa) has a long history of consumption by Ecuadorian natives in regions where the plant is indigenous. The tea contains the methylxanthines caffeine and theobromine as well as chlorogenic acids, flavonoids, and sugars. Various studies were performed to evaluate the general and genetic toxicology of a standardized liquid concentrate of guayusa (GC). Guayusa concentrate was found to be negative in in vitro genotoxicity tests including the Ames test and a chromosome aberration study in human lymphocytes. The oral median lethal dose (LD50) of GC was >5,000 mg/kg for female rats. Guayusa concentrate was administered to male and female rats in a 90-day subchronic study at 1,200, 2,500, and 5,000 mg/kg/d of GC and a caffeine-positive control at 150 mg/kg/d corresponding to the amount of caffeine in the high-dose GC group. Effects observed in the GC-treated groups were comparable to those in the caffeine control group and included reductions in body weights, food efficiency, triglycerides values, and fat pad weights and increases in blood chemistry values for serum aspartate aminotransferase, serum alanine aminotransferase, and cholesterol and adaptive salivary gland hypertrophy. No signs of incremental toxicity due to any other components of guayusa were observed. The studies indicate no harmful effects of GC in these test systems.