Bixa orellana L. Derived Phytochemicals against Alcohol Dehydrogenase of Escherichia coli

Phytochemicals from Bixa orellana L.plant extract are traditionally used to cure Jaundice. It is caused by Escherichia coli. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that benzoic acid can effectively deactivate the alcohol dehydrogenase enzyme thereby interrupting the life cycle of the organism

Extracción de compuestos fenólicos y actividad antioxidante de hojas de Bixa orellana L. (achiote) / Extraction of phenolic compounds and antioxidant activity from leaves of Bixa orellana L. (achiote)

Introducción: Bixa orellana L. también conocida como achiote, es una planta con un rico depósito de compuestos bioactivos, tales como, saponinas, alcaloides y flavonoides, a los cuales se les atribuyen propiedades fisiológicas entre las que se encuentran: actividad antimicrobiana, antiinflamatoria, antialérgica, antiviral, anticancerígena, antioxidante, entre otras. Objetivos: evaluar el efecto del tiempo de extracción y la relación solvente/ sobre el contenido de fenoles totales, así como el efecto del contenido de sólidos y el pH de la solución, sobre la actividad antioxidante del extracto de hojas de B. orellana. Métodos: el contenido total de fenoles fue evaluado por el método de Folin-Ciocalteu. La actividad antioxidante se determinó por los métodos espectrofotométricos de reacción con el radical 2,2´-azino-bis (3-etilbenzotiazolin-6- ácido sulfónico) y la medida de la capacidad reductora sobre el Fe+3, los resultados se expresaron como micromoles de equivalentes Trolox por gramo de extracto (µmol ET.g-1). Resultados: las condiciones del proceso que más favorecen la extracción de compuestos fenólicos desde las hojas de Bixa orellana L. son: tiempo de extracción de 60 h y relación solvente/ hojas (v/p) de 4/1. El contenido máximo de fenoles totales fue de 144,77 ± 9,66 mgAT.g-1, que al someterlo a una solución de pH de 8 y 11,7 °Brix, presenta una actividad antioxidante de 4406,83 ± 43,30 µmol ET.g-1 por el método de reacción con el radical 2,2´-azino-bis (3-etilbenzotiazolin-6- ácido sulfónico) y 4547,22 ± 53,19 µmolET.g-1 por el método de medida de la capacidad reductora sobre el Fe+3. Conclusiones: se demostró que la cantidad de fenoles totales extraídos de hojas de B. orellana dependen de la relación solvente/material vegetal y del tiempo de extracción; asimismo, se encontró que el pH tiene efecto sobre la actividad antioxidante determinada por la reacción con el radical 2,2´-azino-bis (3-etilbenzotiazolin-6- ácido sulfónico) y la medida de la capacidad reductora sobre el Fe+3(AU)

Introduction: Bixa orellana L. also known as annatto, is a plant with a high content of bioactive compounds, such as saponins, alkaloids and flavonoids that have physiological properties as antimicrobial, anti-inflammatory, anti-allergenic, antiviral, anticarcinogenic and antioxidant properties, among others. Objectives: To evaluate the effect of extraction time and the solvent/ leaf ratio over the total phenolic content and the effect of solids and pH of the solution on the antioxidant activity in extracts from Bixa orellana leaves. Methods : The ethanol extract of leaves of Bixa orellana was obtained by percolation and filtering. Total phenol content was evaluated by the Folin-Ciocalteu. The antioxidant activity was determined by spectrophotometric methods as reaction with 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) and ferric reducing antioxidant power, the results were expressed as micromoles of Trolox equivalents per gram of extract (µmolTE.g -1). Results: Process conditions that improve the extraction of phenolic compounds from Bixa. orellana extract are: extraction time of 60 h and solvent/ leaves ratio (v/w) of 4/1. The maximum total phenol content was 144.77 ± 9.66mgAT.g-1, which when subject to a solution of pH 8 and 11.7 °Brix, has an antioxidant activity of 4406.83 ± 43.30 µmolTE.g-1 by the 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) reaction and 4547.22 ± 53.19 µmolET.g-1 by ferric reducing antioxidant power. Conclusion: It showed that the amount of TF extracted from leaves of Bixa orellana depend on the solvent/ leaves ratio and the extraction time. In addition, it found that the pH has an effect on the antioxidant activity determined by the methods 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) reaction and ferric reducing antioxidant power(AU)

Antibacterial activity of Bixa orellana L. (achiote) against Streptococcus mutans and Streptococcus sanguinis

Objective: To evaluate the cytotoxic and antibacterial effect of Bixa orellana L. (B. orellana) (achiote) methanol extract against Streptococcus mutans (ATCC 25175) (S. mutans) and Streptococcus sanguinis (ATCC 10556) (S. sanguinis). Methods: Two methanol extracts of B. orellana were prepared in vitro, from the seeds and leaves. The antibacterial activity of extracts against S. mutans and S. sanguinis was evaluated using the cup-plate agar diffusion method. The minimum inhibitory concentration (MIC) was determined using the microdilution method and the cytotoxic activity was determinated by using the cell line MDCK. Results: A stronger antibacterial effect was observed with the leaves methanolic extract with an inhibition zone of (19.97 ± 1.31) mm against S. mutans and (19.97 ± 1.26) mm against S. sanguinis. The methanolic extract of the seeds had an activity of (15.11 ± 1.03) mm and (16.15 ± 2.15) mm against S. mutans and S. sanguinis, respectively. The MIC of the leaf and the seed extracts against S. sanguinis was 62.5 and 125 μg/mL, respectively, and the MIC of the leaf extract against S. mutans was 62.5 μg/mL, and for the seed extract it was 31.25 μg/mL. The 50% cytotoxic concentration was 366.45 and 325.05 μg/mL for the leaves and seeds extracts, respectively. Conclusions: The experimental findings demonstrated the antibacterial effect of the methanolic extract of B. orellana (achiote) on S. mutans and S. sanguinis. The extract of this plant is cytotoxic at high concentrations.

Análise preliminar da citotoxicidade dos aditivos alimentares urucum e cúrcuma / Cytotoxicity of the food additive annatto and turmeric

Este trabalho teve por objetivo avaliar a ação dos aditivos alimentares urucum e cúrcuma em células meristemáticas de pontas de raízes de Allium cepa nas doses de 5 e 10 g, nos tempos de exposição de 24 e 48 horas. Utilizou-se para cada dose um grupo de cinco bulbos de cebolas, que primeiramente foram enraizados em água destilada, e em seguida transferidos para as suas respectivas doses. Analisaram-se células em todo ciclo celular, totalizando 5.000 para cada controle e tempo de exposição. Os dados obtidos foram submetidos à análise estatística do Qui-quadrado (p<0,05). A partir dos resultados observou-se que as doses do urucum e do cúrcuma tiveram efeito antiproliferativo significativo sobre o ciclo celular deste sistema-teste. Também foi verificado número significativo de aberrações celulares nos dois tempos de exposição avaliados de todas as doses estudadas. Portanto, nas condições analisadas, o urucum e o cúrcuma mostraram-se citotóxicos e genotóxicos.

This study aimed to evaluate the effect of food additives annatto and turmeric tip cells of Allium cepa roots in doses of 5 or 10 g in exposure times of 24 and 48 hours. A group of five onion bulbs was used for each dose. Each dose was first embedded in distilled water and then transferred to their respective doses. Cells were analyzed throughout the cell cycle, totaling 5000 for each control and exposure time. The obtained data were subjected to statistical analysis Chi-square (p <.05). From the results it was observed that the doses of turmeric and annatto had significant antiproliferative effect on the cell cycle of this test system. They also found a significant number of cellular aberrations in the two exposure times evaluated all doses studied. Therefore, under the conditions studied, annatto and turmeric proved cytotoxic and genotoxic.

Secagem solar de grãos residuais e farinha de grãos residuais de urucum / Solar drying of annatto grains and waste grains flour of annatto

O subproduto constituído pelos grãos residuais de urucum é resultante do processo de extração da bixina, principalmente na indústria alimentícia, utilizando-se como meio extrator óleo vegetal. Atualmente, a maioria desses grãos vem sendo descartada, no entanto vê-se a possibilidade de incorporá-los na dieta humana, uma vez que, como todos os grãos, são ricos em carboidratos, proteínas e fibras alimentares. Neste trabalho objetivou-se secar, em um secador solar, grãos residuais e farinha de grãos residuais de urucum, com e sem a camada de óleo proveniente do processo de extração industrial da bixina, ajustar diferentes modelos matemáticos as curvas de secagem e determinar a difusividade efetiva das amostras. Durante as secagens solares foi verificado, no interior do secador, temperaturas médias superiores a 50 °C. Observou-se que as amostras de grãos residuais de urucum com e sem a camada de óleo demoram um maior intervalo de tempo para atingirem o teor de água de aproximadamente 5% b.u., enquanto que as amostras de farinhas de grãos residuais de urucum com e sem óleo tiveram as maiores taxas de secagens. Todos os modelos matemáticos avaliados representaram de forma satisfatória a cinética de secagem das amostras testadas, com valores de R2 superiores a 0,96 e valores de DQM inferiores a 1,0, sendo que o modelo de Midilli foi o que apresentou os melhores parâmetros de ajustes. A difusividade efetiva apresentou-se na ordem de 10-11 m2 s-1, para todas as amostras estudadas, sendo que o maior valor foi encontrado para a amostra de grão sem óleo (6,47 × 10-11 m2 s-1) e o menor para a farinha sem óleo (0,69 × 10-11 m2 s-1).

The byproduct consisting of annatto grains is waste resulting from the extraction of bixin, especially in the food industry, using as extractant vegetable oil. Currently, most of these grains has been ruled out, however sees the possibility of incorporating them in the human diet, since, like all grains are rich in carbohydrates, protein and dietary fiber. The objective of this study was to dry annatto grains and waste grains flour of annatto, with and without the oil layer from the industrial process of extraction of bixin, in a solar dryer and fit different mathematical models of the drying and determine the diffusivity effective samples. During the solar drying was observed inside the dryer, average temperatures exceeding 50 °C. It was observed that samples of grain annatto with and without oil layer takes a longer time to achieve moisture content of approximately 5% wb, whereas the samples of waste grains flour of annatto with and without oil had the highest rates of drying. All mathematical models evaluated represent satisfactorily the drying kinetics of the samples tested, with R2 values greater than 0.96 and DQM values below 1.0, and the Midilli model presented the best parameters adjustments. The effective diffusivity presented in the order of 10-11 m2 s-1 for all samples, while the highest value was found for the sample of grain without oil (6.47 × 10-11 m2 s-1) and lowest for flour without oil (0.69 × 10-11 m2 s-1).

Analysis of water soluble polysaccharides as a potential chemotaxonomic marker for landraces in Bixa orellana.

Annatto tree (Bixa orellana L.) is native to Brazil and is now under cultivation in many parts of world for its reddish orange ‘annatto’ dye. There are three types of landraces in annatto and they are distinguished based on fruit shape i.e., ovate, conical and hemispherical, whose pigment yield differs. Since annatto pigment yield varies with landrace, it is necessary to characterize markers towards the identification of landraces. In this study, we characterized water soluble polysaccharides (WSP) of twigs from three landraces using size-exclusion chromatography (SEC), fourier-transform Infrared spectroscopy (FT-IR), nuclear magnetic resonance spectroscopy (NMR) and gas liquid chromatography (GLC) for their potential use as chemotaxonomic markers to distinguish the landraces. GLC analysis on WSP showed hemispherical type contained 38% rhamnose, while conical and ovate types contained 17% and 34% glucose, respectively. Thus, glucose and rhamnose content of WSP could be used to distinguish the three landraces. Further, differences in calculated molecular weight as revealed by SEC (281.8, 151.3 and 79.4 kDa for conical, hemispherical and ovate types, respectively) could also be used to distinguish the three landraces.

Bacterial growth inhibition potential of annatto plant parts

Objective: To study the antibacterial efficacy of Bixa orellana leaves and deseeded fruit capsule extracts against both Gram positive and Gram negative bacteria. Methods: The antibacterial activity of the ethanolic, methanolic, acetone and dimethyl sulphoxide extracts of B. orellana were tested against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis,Bacillus cereus and Staphylococus aureus by disc diffusion method. Results: The antibacterial activity of leaf was more pronounced even at low concentrations and fruit extracts exhibited the same at relatively higher concentrations. Only DMSO extract of seeds showed growth inhibition of S. aureus, B. subtilis, B. cereus, and P. aeruginosa. Conclusions: The present study suggested that the leaves and deseeded capsule extracts of B. orellana possess significant antibacterial activity thereby providing substantial support for the ethanobotanical applications of this plant.

Leaf and Seed extracts of Bixa orellana L. exert anti-microbial activity against bacterial pathogens.

Methanol extracts of the leaf & seed of Bixa orellana L. were studied for in vitro antimicrobial activity against MTCC strains of Staphylococcus aureus, Salmonella typhi, Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterococcus fecalis, Vibrio cholera, Moraxella catarrhalis, Acinetobacter sp., Brucella sp. along with fungal pathogens Candida albicans, Aspergillus niger and the dermatophytes Trichophyton mentagrophytes & Trichophyton rubrum. Leaf extract of B. orellana at 1000 μg/ml concentration showed significant inhibition against all the tested bacteria and fungus, with highest inhibition zone (18±0.3 mm) against S. typhi, Acinetobacter sp., T. mentagrophytes and T. rubrum. Seed extract of B. orellana was comparitively less efficacious in most of the tested pathogens, except Brucella sp. which was appreciably inhibited (15±0.1 mm). Minimum Inhibitory Concentration (MIC) of leaf extract was determined as 15.62 μg/ml against S. aureus and 31.25 μg/ml for K. pneumoniae, P. aeruginosa, E. fecalis & S. typhi, on average. Among the dermatophytes, 78.2% inhibition was seen in T. mentagrophytes & T. rubrum. Scanning Electron Microscopic (SEM) studies of the treated P. aeruginosa cells revealed disintegration & aggregation of cells after treatment with the leaf extract. Phytochemical analysis of leaf & seed extracts suggested the presence of flavonoids, tannins, saponins, steroids. Alkaloids were detected only in the leaf & anthroquinones, in the seeds.

La Bixa orellana L en el tratamiento de afecciones estomatológicas, un tema aún por estudiar / The Bixa orellana L in treatment of stomatology affections: a subject that hasn't studied yet

De las semillas de la planta tropical Bixa orellana L (bija) se extrae un pigmento (el annatto) que està compuesto bßsicamente por carotenoides (bixina/nor-bixina). Por su resistencia a la acción de los agentes químicos, pero no a los efectos del sol, se emplea con preferencia para colorear alimentos y bebidas. La dosis diaria permitida recomendada por la OMS es de 0-0,065 mg/kg de peso corporal. El annato es un colorante con muy baja toxicidad para el humano, reportàndose escasos incidentes de eventos alérgicos. A los extractos de bija le han sido demostrados diversas propiedades farmacológicas, entre las cuales se encuentran: antitumoral, antiinflamatoria, astringente, emoliente, antiséptica, antibacteriana, antioxidante, cicatrizante entre otras y se ha indicado en el tratamiento de infecciones bacterianas de forma general, estomatitis, y en la curación de heridas y quemaduras. Teniendo en cuenta que en el arsenal terapéutico al alcance del estomatólogo son escasos los productos de acción local cicatrizante, antioxidante, antiinflamatoria y antimicrobiana y mucho menos un producto que reúna todas estas propiedades juntas, es que se procedió a realizar una actualización bibliogràfica del tema Bixa orellana L, referido a su composición química, toxicología y efectos farmacológicos de interés para el tratamiento potencial de desórdenes en el campo de la estomatología. Existen evidencias de la utilidad de estos extractos para el tratamiento de desórdenes estomatológicos, pero estas deben aún ser validadas desde el punto de vista clínico con ensayos realizados de acuerdo con las buenas pràcticas clínicas, antes de cualquier aplicación en los servicios estomatológicos cubanos.

From the Bixa orellana L (bija) tropical plant seeds a pigment (annatto) was extracted which is basically composed by carotenoids (bixine/nor-bixine). Due to its resistance to chemical agent's action but not to sun effects, it is used preferably to food and drinks coloring. Daily dose allowance recommended by HWO is of 0-0,065 mg/kg/bw. Annate is a coloring agent with a very low toxicity for humans, even when some low rates of allergic events have been reported. Annate extracts have different pharmacological properties including the antitumor, anti-inflammatory, astringent, emollient, antiseptic, antibacterial, antioxidant, cicatrizant among others and has been prescribed in wounds and burns cure. Considering that in the therapeutical array available for stomatologist the cicatrizant antioxidant, anti-inflammatory and antimicrobial local actions are limited and by no means a product fulfilling all these properties together, we made a bibliographic updating of Bixa orellana L subject related to its chemical, toxicology composition and interesting pharmacological effects for potential treatment of stomatology disorders. There are evidences on usefulness of these extracts for stomatology disorders, but these must to be still validated from the clinical point of view with trials performed according to the good clinical practices before any application in Cuban stomatology services.

La Bixa orellana L como posible sustancia reveladora de placa dentobacteriana / Bixa orellana L: a potential substance for detection of dentobacterial plaque

En la actualidad las plantas naturales se han convertido en una importante fuente de elaboración de medicamentos, lo que nos ha motivado a realizar una revisión bibliográfica sobre la Bixa orellana L. Este es un producto natural fácilmente asequible en nuestro medio por encontrarse ampliamente distribuido en nuestro territorio, y apoyándonos en sus ya conocidas propiedades de tinción en alimentos, surge la idea de su posible uso estomatológico como sustancia reveladora de placa dentobacteriana. Es objetivo del trabajo ampliar conocimientos sobre las propiedades y uso de Bixa orellana L en los seres humanos. Se revisaron artículos originales y de revisión localizados mediante Pub-Med, Google, revistas internacionales y nacionales reconocidas. Se ha demostrado que la Bixa no provoca alteraciones patológicas en el ser humano y tiene importantes propiedades: antifúngicas, antipiréticas, antibacterianas, antiinflamatorias y antigonorreicas, entre otras, y se utiliza en la tinción de alimentos como: mantequilla, quesos y aceites. Con esta propuesta pretendemos incrementar las actividades de promoción y prevención de la caries dental y las periodontopatías, elevar el nivel de educación para la salud dental en nuestra población, disminuir los tratamientos curativos, prolongando así la salud de los dientes remanentes(AU)

At the present time natural plants become a significant drug manufacture source, thus, we made a bibliographic review on Bixa orellana L. This is a natural product easily accessible since it is fully distributed in our country, and due to its known dyeing properties in foods, arise the idea of its potential Stomatology use as a revealing substance of dentobacterial plaque. To expand our knowledges on the properties and use of Bixa orellana L in human being. We reviewed the original papers retrieved by Pub-Med, Google, and recognized international and national journals. It has been demonstrated that Bixa does not provokes pathologic alterations in human being and also has significant antifungal, antipyretic, antibacterial, anti-inflammatory, and anti-gonorrhea properties among others, and it is used in food dyeing: butter, cheeses, and oils. With this proposal, we try to increase the promotion and prevention activities of dental caries and the periodontal diseases, to raise the educational level for dental hygiene in our population, to decrease the curative treatments, thus extending the remnant teeth health(AU)

Germinação de sementes de urucu em diferentes temperaturas e substratos / Germination of annatto seeds under different temperatures and substrates

Objetivou-se, neste trabalho, analisar o comportamento germinativo das sementes de urucu cultivar Casca Verde, com e sem escarificação, sob regime de diferentes temperaturas e substratos. O trabalho foi realizado no Laboratório de Tecnologia e Análise de Sementes do Centro de Ciências Agrárias da Universidade Federal do Espírito Santo (CCA-UFES). O delineamento experimental utilizado foi o inteiramente casualizado, num esquema fatorial 2x6x4 (dois tratamentos físicos nas sementes, seis substratos e quatro temperaturas), totalizando 48 tratamentos, com quatro repetições de 50 sementes. Os tratamentos físicos foram: as sementes intactas e as sementes escarificadas; os substratos foram: a areia, a vermiculita, a fibra de coco, o pó de serra, o Plantmax e o rolo de papel Germitest ; e, as temperaturas testadas foram constantes de 20, 25 e 30°C e alternada de 20-30°C. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey. Os resultados obtidos evidenciaram maior porcentagem de germinação das sementes de urucu, semeadas nos substratos areia, vermiculita e rolo de papel; as temperaturas de 25, 30 e 20-30°C foram mais adequadas para testes de germinação dessas sementes.

This work was carried out with the objective to verify the effect of temperature and substrate on germinative capacity of annatto seeds. This study was developed in the Laboratories of Seed Analysis of Agrarian Science Center that belongs to the Universidade Federal do Espirito Santo (CCA-UFES), located in Alegre ES, Brazil. The experimental design was 2x6x4 factorial involving: (i) two treatments in the seeds, (ii) six substrates, and (iii) four temperatures. Four replications were realized using 50 seeds at each experimental unit. Treatments refer to intact and scarified seeds. Substrates utilized were sand, vermiculite, coconut fiber, wood fiber, Plantmax and paper roll. Temperatures employed were 20, 25, 30 and 20-30°C. Average contrasts were verified using Tukey test at 5 percent of significance. It can be concluded: (i) that better percentages of germination occurred by using sand substrate, vermiculite and paper roll; and (ii) temperatures of 25, 30 and 20-30°C can be utilized in germination tests of annatto seeds.

Actividad analgésica del extracto acuoso liofilizado de las hojas de Bixa orellana L. en ratones albinos / Analgesic activity of the lyophilized aqueous extract of the leaves of Bixa orellana L. in albino mice

Se realizó un estudio experimental con el objetivo de evaluar el efecto analgésico del extracto acuoso liofilizado de las hojas de Bixa orellana L, a dosis de 100 y 150 mgkg-l administrado por vía oraL Para el desarrollo del mismo se utilizaron los Métodos de "Hot-plate" y "Contorsiones abdominales". Los animales seleccionados fueron ratones albinos machos de la cepa Balb/C53, con un peso corporal comprendido entre 23.67+ l.03g. Se utilizaron como controles positivos paracetamol e indometacina a dosis de 400 mgkg-l y 10 mgKg-l, respectivamente. Bixa orellana L. presenta actividad analgésica a nivel periférico, en el modelo de contorsiones abdominales, directamente proporcional a la dosis, bajo las condiciones experimentales.

Indução de brotações em explantes de segmentos de folhas de plântulas de urucueiro em diferentes citocininas / Induction of shoot buds in leaf explants of annatto seedlings in differents cytokinins

Segmentos de folhas de plântulas de urucum (Bixa orellana L), foram cultivados in vitro, em um meio básico idealizado por Murashige & Skoog (1962), suplementado com 0,5mg/l dos reguladores do crescimento, benzilaminopurina (BAP), cinetina (KN) e isopentenil-adenina (2iP), isolados ou combinados entre si, com o objetivo de avaliar qual a melhor citocinina ou combinação destas para indução de brotações de boa qualidade visando à micropropagação. Após 60 dias de cultivo, as gemas obtidas foram transferidas para um meio MS modificado, contendo a metade da formulação de sais minerais, sem a suplementação de reguladores do crescimento. Gemas de melhor qualidade foram obtidas nos tratamentos onde a KN e o BAP estavam em combinação. A KN sozinha não foi eficiente na indução de brotações. O maior número de gemas por explante foi obtido com KN, BAP e 2iP e no tratamento com apenas BAP.

Excised leaf explants of annatto (Bixa orellana L.) seedlings were cultivated on a Murashige & Skoog basal medium containing benzylaminopurine (BAP), kinetin (KN) and isopentenyl-adenine (2iP) individually or in combination, each at concentration of 0.5mg/l, with the purpose of evaluated the best cytokinins or cytokinins combination for shoot bud formation. After 60 days, regenerated shoots were transferred to half strenght MS medium without growth regulators. Best shoots were observed where KN and BAP were added in combination. Isolated KN was ineffective for shoot bud formation. The maximum average number of shoot buds per leaf explant was obtained on medium with BAP, KN and 2iP in combination and on medium supplied of isolated BAP.