Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp / In vitro antimicrobial activity of Bixa orellana L. leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp.

Objetivos . Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. Materiales y métodos . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. Resultados . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). Conclusiones . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

Bixinoids Derived from Bixa orellana as a Potential Zika Virus Inhibitor Using Molecular Simulations. Antiviral Effect on the Zika Virus of Bixinoids

Abstract Zika fever is a viral infection of great relevance in public health, especially in tropic regions, in which there is a predominance of mosquitoes of the genus Aedes, vectors of the disease. Microcephaly in neonatal children and Guillain-Barré syndrome in adults can be caused by the action of the Zika virus (ZIKV). Non-structural proteins, such as NS2B, NS3 and NS5, are important pharmacological targets, due to their action in the life cycle. The absence of anti-Zika drugs raises new research, including prospecting for natural products. This work investigated the in silico antiviral activity of bixin and six other derived molecules against the Zika viral proteins NS2B-NS3 and NS5. The optimized structure was subjected to molecular docking to characterize the interaction between bixinoids and ZIKV non-structural proteins, where significant interactions were observed with amino acid residues in the catalytic site in each enzyme. These results suggest that bixin and ethyl bixin has the potential to interfere with the enzymatic activity of NS2B, NS3 and NS5, thus being an indication of being a promising anti-Zika agent.

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

Bixa orellana Derived Phytochemicals against Entamoeba histolytica Causing Dysentery

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

Nutritional value and inclusion levels of residual annatto seed meal in diets for Japanese quails / Valor nutricional y niveles de inclusión de harina residual de semilla de achiote en dietas para codorniz Japonesa / Valor nutritivo e níveis de inclusão do farelo residual da semente de urucum em dietas para codornas Japonesas

Abstract Background: Residual annatto seed meal (RASM) is a byproduct after extraction of bixin in the cosmetic industry and may be used in animal diets. Objective: Two experiments were carried out to determine the nutritive value and effect of RASM inclusion levels in the diets of Japanese quails. Methods: For the digestibility experiment, 160 Japanese quails were distributed in a randomized study with two treatments and 10 replicates with eight birds each. The treatments consisted of a reference diet and a test diet containing 75% of the reference diet and 25% of RASM. For the performance assay, 192 Japanese quails were used in a randomized study with six treatments and four replicates with eight birds each. Treatments consisted of a control corn-based diet (Contr1) and another diet based on sorghum + canthaxanthin (Contr2), and four diets based on sorghum with increasing inclusion levels of RASM (0, 3, 6, and 9%). Results: Digestibility of dry matter and crude protein, metabolization coefficient of gross energy, and retention of calcium and phosphorus were 41.05, 43.12, 47.97, 25.49, and 30.35%, respectively. RASM inclusion did not influence bird productivity; however, feed conversion worsened in treatments with 0-6% RASM inclusion relative to that in Contr1. Yolk color increased due to RASM treatment. Conclusion: RASM may be included in sorghum-based diets for quails at 9%, with improvement in yolk color and no negative effects on productivity and egg quality.

Resumen Antecedentes: La harina de la semilla de achiote (RASM) es un subproducto de la extracción de bixina por la industria cosmética y puede ser usada en alimentación animal. Objetivo: Se realizaron dos experimentos para determinar el valor nutritivo y los efectos de la inclusión de RASM en dietas para codorniz. Métodos: Para el experimento de digestibilidad, 160 codornices japonesas fueron distribuidas en un estudio al azar con dos tratamientos y 10 repeticiones con ocho aves cada uno. Los tratamientos consistieron en una dieta referencia y una dieta de prueba, la cual estuvo compuesta por 75% de la dieta referencia y 25% de RASM. Para el ensayo de desempeño, 192 codornices japonesas fueron usadas en un estudio al azar con seis tratamientos y cuatro repeticiones con ocho aves cada uno. Los tratamientos consistieron en una dieta control a base de maíz (Contr1) y otra a base de sorgo + cantaxantina (Contr2) y cuatro dietas a base de sorgo con niveles de inclusión de RASM (0, 3, 6 y 9%). Resultados: Los coeficientes de digestibilidad de materia seca y proteína bruta, el coeficiente de metabolización de la energía bruta y la retención de calcio y fósforo fueron 41,05; 43,12; 47,97; 25,49 y 30,35%, respectivamente. La inclusión de RASM no influyó la productividad de las aves; entretanto la conversión alimenticia empeoró debido a los tratamientos con 0-6% RASM comparado con el tratamiento Contr1. El color de la yema aumentó debido a la RASM. Conclusión: RASM puede ser incluido en dietas a base de sorgo para codornices en un 9%, debido a la mejora en el color de la yema, además de no tener ningún efecto negativo en el desempeño productivo y calidad del huevo.

Resumo Antecedentes: O farelo residual da semente de urucum (RASM) é um subproduto da extração da bixina pela indústria cosmética e pode ser usado na alimentação animal. Objetivo: Dois experimentos foram conduzidos para determinar o valor nutritivo e os efeitos de inclusão do FRSU em dietas para codornas Japonesas. Métodos: Para o experimento de digestibilidade, 160 codornas Japonesas foram distribuídas em um estudo casualizado com dois tratamentos e 10 repetições com oito aves cada. Os tratamentos consistiram em uma dieta referência e uma dieta teste, a qual era composta por 75% da dieta referência e 25% de RASM. Para o ensaio de desempenho, 192 codornas Japonesas foram usadas em estudo ao acaso com seis tratamentos e quatro repetições com oito aves cada. Os tratamentos consistiram em uma dieta controle a base de milho (Contr1) e outra à base de sorgo com cantaxantina (Contr2) e quatro dietas a base de sorgo com níveis de inclusão de RASM (0, 3, 6 e 9%). Resultados: Os coeficientes de digestiblidade de matéria seca e proteína bruta, coeficiente de metabolização da energia bruta e retenção de cálcio e fósforo foram 41,05; 43,12; 47,97; 25,49 e 30,35%, respectivamente. A inclusão do FRSU não influenciou a produtividade das aves; entretanto, a conversão alimentar piorou devido aos tratamentos com 0-6% FRSU comparado com o tratamento Contr1. A cor da gema aumentou devido ao FRSU. Conclusão: RASM pode ser incluido em dietas baseadas em sorgo para codornas em 9%, devido a mehoria na cor da gema e nenhum efeito negativo no desempenho produtivo e qualidade de ovo.

Assessment of analgesic and neuropharmacological activity of leaves of Bixa orellana (Family: Bixaceae)

Background:Bixa orellanaLinn, (Family: Bixaceae) commonly known as “Lipstick tree” have been extensively used traditional medicine in India and others part of the world to cure laxative, cardiotonic, hypotensive,expectorant, antibiotic, antipyretic, aphrodisaic etc. It has been found that B. orellanacontains different phytochemical groups such as phenolic compounds, glycosides, tannins, steroids, alkaloids, saponins.The project work has been designed to investigate the phytochemical nature (group determination of plant constituents), analgesic and neuropharmacological activity of B.orellanaleaves extract. Methods:Analgesic activity of B. orellanawas determined by acetic acid induced writhing, hot plate, tail immersion and paw tickling test/formalin induced nociceptortest. Open field, hole cross, tail suspension and light/dark box test were employed for the assessment of neuropharmacological activity of B. orellana.Results:Phytochemical screening revealed the presence of reducing sugar, alkaloids, glycosides, gum,terpenoids, tannins, steroids and flavonoids. In analgesic activity test, the sample showed significant analgesic activity. The dose-dependent neuropharmacological activity is shown in neuropharmacological activity test.Conclusions:Our exploration suggests that B. orellanacontains bioactive compounds and it should be studied further for isolation and purification of such novel compounds

The annatto carotenoids and the norbixin absorption coefficient / Carotenoides de urucum e o coeficiente de absorção danorbixina

Annatto seeds present cis-bixin as the major carotenoids, but the norbixin salt is the main pigment present in dyes obtained from the alkaline extraction process. For analyzing the norbixin, the absorptions are obtained in the two spectral peaks with higher intensity, but discrepancies in the published extinction values have led to serious doubts. Taking into account the use of absorption coefficient for evaluating the norbixin concentration in annatto seeds and extracts, the present study evaluated the absorption coefficient in various solvents and the total uncertainty associated with this value was determined. The norbixin standard was prepared from annatto seeds, purified by column chromatography and the purity of norbixin was evaluated by HPLC-DAD. The absorption coefficients were determined for norbixin in different solvents and the uncertainty was evaluated. The 0.5% potassium hydroxide, the main solvent used for marketing the annatto seeds and extracts, showed the absorption coefficients of 2887 at 454nm and of 2546 at 483nm, and the estimation of expanded uncertainty (K=2) was ±86 and 85 g 100 mL-1, respectively.

O principal carotenoide em sementes de urucum é a cis-bixina, mas o sal de norbixina torna-se o principal pigmento presente em corantes obtidos pelo processo de extração alcalino.Para a análise de norbixina as absorbâncias são obtidas nos dois picos espectrais de maior intensidade, mas discrepâncias no valor de absortividade levam a sérias dúvidas. Levando-se em conta o uso do coeficiente de absorção na avaliação da concentração de norbixina em sementes de urucum e extratos, este trabalho teve como objetivo avaliar o coeficiente de absorção em vários solventes e determinar a incerteza expandida associada a esse valor. Para a execução do estudo, um padrão de norbixina foi preparado a partir de sementes de urucum, purificado em coluna aberta e a pureza da norbixina foi avaliada por HPLC-DAD. O coeficiente de absorção da norbixina foi determinado pela construção de curvas analíticas em diferentes solventes e a incerteza expandida foi avaliada. O hidróxido de potássio a 0,5%, principal solvente utilizado na comercialização de sementes de urucum e extratos, apresentou o coeficiente de absorção determinado em 2887 a 454nm e 2546 a 483nm e a estimação da incerteza expandida (K=2) foi86 e 85 g 100 mL-1, respectivamente.

The annatto carotenoids and the norbixin absorption coefficient / Carotenoides de urucum e o coeficiente de absorção da norbixina

Annatto seeds present cis-bixin as the major carotenoids, but the norbixin salt is the mainpigment present in dyes obtained from the alkaline extraction process. For analyzing the norbixin,the absorptions are obtained in the two spectral peaks with higher intensity, but discrepanciesin the published extinction values have led to serious doubts. Taking into account the use ofabsorption coefficient for evaluating the norbixin concentration in annatto seeds and extracts, thepresent study evaluated the absorption coefficient in various solvents and the total uncertaintyassociated with this value was determined. The norbixin standard was prepared from annattoseeds, purified by column chromatography and the purity of norbixin was evaluated by HPLC-DAD.The absorption coefficients were determined for norbixin in different solvents and the uncertaintywas evaluated. The 0.5% potassium hydroxide, the main solvent used for marketing the annattoseeds and extracts, showed the absorption coefficients of 2887 at 454nm and of 2546 at 483nm,and the estimation of expanded uncertainty (K=2) was ±86 and 85 g 100 mL-1, respectively.

O principal carotenoide em sementes de urucum é a cis-bixina, mas o sal de norbixina torna-seo principal pigmento presente em corantes obtidos pelo processo de extração alcalino.Para a análise de norbixina as absorbâncias são obtidas nos dois picos espectrais de maiorintensidade, mas discrepâncias no valor de absortividade levam a sérias dúvidas. Levando-se emconta o uso do coeficiente de absorção na avaliação da concentração de norbixina em sementesde urucum e extratos, este trabalho teve como objetivo avaliar o coeficiente de absorção emvários solventes e determinar a incerteza expandida associada a esse valor. Para a execução doestudo, um padrão de norbixina foi preparado a partir de sementes de urucum, purificado emcoluna aberta e a pureza da norbixina foi avaliada por HPLC-DAD. O coeficiente de absorçãoda norbixina foi determinado pela construção de curvas analíticas em diferentes solventes e aincerteza expandida foi avaliada. O hidróxido de potássio a 0,5%, principal solvente utilizadona comercialização de sementes de urucum e extratos, apresentou o coeficiente de absorçãodeterminado em 2887 a 454nm e 2546 a 483nm e a estimação da incerteza expandida (K=2) foi86 e 85 g 100 mL-1, respectivamente.

Actividad antioxidante del complejo de inclusión del extracto de semilla de Bixa orellana en ß-ciclodextrina obtenido por CO2 supercrítico / Antioxidant activity of the inclusion complex of Bixa orellana seeds extract in ß-cyclodextrin obtained by supercritical CO2

Antecedentes: los extractos de Bixa orellana son utilizados como colorante de alimentos, y presentan una actividad antioxidante de importancia farmacéutica. Su uso puede limitarse por su inestabilidad, donde el proceso de inclusión en ß-ciclodextrina (ß-CD) por fluido supercrítico con CO2 (FSC-CO2) es una alternativa para mediar esta desventaja. Objetivos: comparar diferentes condiciones de temperatura y presión de extracción por FSC-CO2, para determinar cuál de estas permite obtener un extracto de Bixa orellana con mayor actividad antioxidante, y evaluar el efecto de la inclusión en ß-CD, en dicha capacidad antioxidante. Métodos: se obtuvieron extractos por FSC-CO2 variando condiciones de presión y temperatura: (I) 3583 psi, 35°C, (II) 1413 psi, 35°C, (III) 2184 psi, 45°C, (IV) 5076 psi, 45°C, y (V) 2300 psi, 40°C. Se evaluó la actividad antioxidante de los extractos obtenidos mediante el método DPPH, determinando su IC50. Se realizó el complejo de inclusión en ß-CD del extracto que presentó la mayor actividad antioxidante, por el método FSC-CO2, y fue caracterizado por IR, DSC y RMN. Mediante análisis comparativo de los espectros de la ß-CD, extracto libre y el complejo extracto/ß-CD, se verificó el acomplejamiento, y se evaluó la capacidad antioxidante del complejo de inclusión. Resultados: el extracto con mayor actividad antioxidante se obtuvo bajo la condición de extracción IV, con un IC50 de 23,55 µg/ mL, seguido del extracto II (28,76 µg/mL), del extracto III (37,23 µg/mL), del extracto V (81,09 µg/mL) y del extracto I (193,82 µg/mL), los cuales presentaron diferencias significativas (P<0,01). Los espectros obtenidos por IR, DSC y RMN presentaron desplazamientos propios del proceso de encapsulamiento. El valor IC50 del complejo extracto/ß-CD fue de 104,84 µg/mL, siendo significativamente mayor al valor obtenido para el extracto puro (23,55 µg/mL). Conclusión: el extracto de Bixa orellana con una actividad antioxidante mayor se obtuvo por fluido supercrítico a 5076 psi de presión y 45°C de temperatura. Las variaciones de los espectros IR, DSC y RMN demuestran la inclusión del extracto en la ß-CD, y los valores de IC50 indican el efecto protector de la ß-CD ante la reacción con el radical DPPH.

Background: Bixa orellana extracts are used to food coloring and it is important in pharmaceutical industry as a potential source of antioxidant activity. The application may be limited because it is unstable, but the process of inclusion in ß-cyclodextrin (ß-CD) by supercritical fluid CO2 (FSC-CO2) is an alternative to mediate this disadvantage. Objectives: we compared different conditions of temperature and pressure of FSC-CO2 extraction, to obtain a Bixa orellana extract with excellent antioxidant activity, and we evaluated the effect of inclusion in ß-CD in the antioxidant capacity. Methods: extracts were obtained by FSC-CO2 with different conditions of pressure and temperature: (I) 3583 psi, 35°C, (II) 1413 psi, 35°C, (III) 2184 psi, 45°C, (IV) 5076 psi, 45°C, and (V) 2300 psi, 40°C. The antioxidant activity by DPPH assay was determined and the IC50 was evaluated. We performed the inclusion complex in ß-CD with the highest antioxidant activity extract. The extract/ß-CD complex was characterized by IR, DSC and NMR. This complexation was verified by comparative analysis of the spectra of the ß-CD, free extract and ß-CD/extract complex. The antioxidant capacity of this inclusion complex was evaluated. Results: the extract with highest antioxidant activity was obtained under the extraction condition IV, with an IC50 of 23.55 µg/mL, followed extract II (28.76 µg/mL), extract III (37.23 µg/mL), extract V (81.09 µg/mL) and extract I (193.82 µg/mL). Analysis showed significant differences (P<0.01) between these extracts. The spectra obtained by IR, DSC and NMR evidence the encapsulation process. The IC50 value of the extract/ß-CD complex (104.84 µg/mL) was significantly higher than the value obtained for the pure extract (23.55 µg/mL). Conclusions: the highest antioxidant activity of Bixa orellana extracts was obtained by supercritical fluid pressure at 5076 psi and temperature of 45°C. Variations in IR, DSC and NMR spectra showed the inclusion of the ß-CD/extract, and the IC50 values indicated the protective effect of ß-CD to the reaction with DPPH radical.

Ação cicatrizante da nanopartícula de prata com norbixina em queimaduras / Healing action of silver nanoparticle with norbixin on burns

Introdução: A queimadura é uma injúria que altera a integridade da pele, decorrente de traumas térmicos. Dentre os compostos não convencionais utilizados na cicatrização de feridas estão a norbixina e as preparações com prata. Objetivo: avaliar a ação cicatrizante da nanopartícula de prata associada à norbixina em queimaduras de Mus musculus. Métodos: Inicialmente, preparou-se uma solução nanoprata/norbixina e a partir desta obteve-se o gel teste. Posteriormente dividiu-se os animais em 3 grupos, sendo negativo (sem tratamento), positivo (sulfadiazina de prata) e teste (gel teste). Nestes animais houve a indução das queimaduras. No 7º e 14º dia realizou-se a eutanásia e os tecidos lesionados foram coletados para análise microscópica. Resultado: Verificou-se potencial vantagem do grupo teste em relação ao negativo e igualdade comparada ao positivo. Conclusão: A nanopartícula de prata associada à norbixina representa uma fonte alternativa para a cicatrização de queimaduras. (AU)

Introduction: Burning is an injury that alters the integrity of the skin, due to thermal trauma. Among the non-conventional compounds used in wound healing are norbixin and silver preparations. Objective: to evaluate the cicatrizing action of silver nanoparticle associated with norbixina in Mus musculus burns. Methods: Initially, a nanoprata / norbixin solution was prepared and from this solution the test gel was obtained. Afterwards the animals were divided into 3 groups, being negative (without treatment), positive (silver sulfadiazine) and test (gel test). In these animals, there was the induction of burns. On the 7th and 14th day, euthanasia was performed and the injured tissues were collected for microscopic analysis. Result: There was a potential advantage of the test group in relation to negative and equality compared to positive. Conclusion: The silver nanoparticle associated with norbixin represents an alternative source for the healing of burns. (AU)

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.

Effects of bixin in high-fat diet-fed-induced fatty liver in C57BL/6J mice

Objective To evaluate the anti-obesity activity of bixin (BIX) on C57BL/6J mice which were fed a high-fat diet (HFD) and to determine the mechanism of this effect. Methods C57BL/6J mice were separately fed a high-calorie diet or a normal diet for 8 weeks, then they were treated with BIX for another 13 weeks. After administration for 13 weeks, the animals were sacrificed. Body adiposity, serum lipid level, and insulin resistance were evaluated. In addition, a histological assay of pancreas and liver, an evaluation of the inhibitory properties on pancreatic lipase, and α-amylase were conducted. Results Administration of BIX significantly decreased the body weight gain, adipocyte size, fat pad weights, hepatic lipid levels in HFD-induced obese mice. In addition, reduced liver weight exhibited decreased serum leptin levels, malic enzyme, glucose-6-phosphate dehydrogenase, hepatic fatty acid synthase, aspartate aminotransferase, alanine aminotransferase and hepatic phosphatidate phosphohydrolase activity. However, superoxide dismutase, catalase, glutathione peroxidase, and glutathione levels were increased in hepatic tissue. BIX also decreased lipid and carbohydrates absorption due to inhibition of pancreatic lipase and α-amylase. Long term supplementation of BIX significantly decreased hyperlipidemia, insulin resistance and glucose level. Decreased levels of hepatic steatosis and the islets of Langerhans appeared less shrunken in HFD-fed mice. Conclusions The antiobesity effect of BIX appears to be associated at least in part, to its inhibitory effect on lipids and carbohydrate digestion enzymes such as pancreatic lipase, α-glucosidase, and α-amylase. The results suggested that BIX also act as an antioxidant and may treat visceral obesity normalizing glucose levels, improving insulin resistance and increasing energy expenditure. Therefore, achiote which has a main component, the carotenoid BIX, could be a viable food for the treatment of obesity and diabetes.

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 concen-tration (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 mg/mL, respectively, and the MIC of the leaf extract against S. mutans was 62.5 mg/mL, and for the seed extract it was 31.25 mg/mL. The 50%cytotoxic concentration was 366.45 and 325.05 mg/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.

Evaluación de propiedades fisicoquímicas de aceite de girasol ( Helianthus annuus ) adicionado con extracto de achiote ( Bixa orellana ) durante calentamiento en horno microondas / Evaluation of physicochemical properties of sunflower oil ( Helianthus annuus ) blended with achiote ( Bixa orellana ) extract during microwave heating

RESUMEN Antecedentes la cocción de alimentos en horno microondas es una tendencia en aumento en la población en general, sin que se tenga en cuenta el efecto sobre su calidad. Objetivo evaluar el efecto del extracto de Bixa orellana sobre aceite de girasol durante el calentamiento en microondas. Materiales y métodos aceite de girasol, con y sin la adición de extracto de Bixa orellana 100 ppm, se sometió a calentamiento durante 12 minutos en horno microondas evaluándose sus propiedades fisicoquímicas a distintos intervalos de tiempo. Resultados la adición de Bixa orellana contribuye a conservar la calidad del aceite de girasol sometido al microondas durante los tres primeros minutos de calentamiento. A tiempos mayores, dadas las temperaturas alcanzadas, no se observa efecto protector. Conclusión la adición de 100 ppm de extracto de Bixa orellana en aceite de girasol permite contrarrestar la oxidación durante tres minutos de calentamiento en microondas.

ABSTRACT Background Microwave cooking is a growing trend in the general population, which does not take into account the effect on food quality. Objective: To evaluate the effect of the extract of Bixa orellana on sunflower oil during microwave heating. Materials and methods Sunflower oil was heated during 12 minutes in a microwave, with and without the addition of 100 mg extract of Bixa orellana, and physicochemical properties were evaluated at different time intervals. Results The addition of Bixa orellana extract contributes to preserve the quality of the sunflower oil during the first three minutes of heating. For longer periods, no protective effect is observed. Conclusion The addition of 100 mg of extract of Bixa orellana on sunflower oil was capable to avoid the oxidation during microwave heating for three minutes.

Desempenho e qualidade dos ovos de poedeiras alimentadas com semente de urucum / Performance and quality of eggs from laying hens fed with annatto seed / Rendimiento y calidad de los huevos de gallinas ponedoras alimentadas con semillas de achiote

Objetivou-se avaliar o efeito da adição de níveis de semente de urucum moída nas rações sobre o desempenho e a qualidade dos ovos de poedeiras comerciais de ovos vermelhos. Os tratamentos foram constituídos por cinco dietas, sendo quatro experimentais contendo níveis de inclusão de semente de urucum moída (0,5; 1,0; 1,5 e 2,0%) e um grupo controle. Adotou-se um delineamento inteiramente casualizado, com quatro repetições e oito aves por unidade experimental. As variáveis analisadas foram: consumo de ração, porcentagem de postura, massa de ovos, conversão alimentar (kg/kg e kg/dz), peso médio do ovo, unidade Haugh, índice de gema, cor, gravidade específica, pH de gema e de albúmen, porcentagem e espessura da casca. As variáveis de desempenho e qualidade dos ovos não foram influenciadas (P>0,05) pelos níveis de semente de urucum estudados, exceto a coloração da gema que apresentou efeito quadrático (Y=5,93+5,86X-1,51X2), demonstrando que a coloração da gema crua foi intensificada com a inclusão de 1,94% de semente de urucum às rações. Conclui-se que adição de semente de urucum moída em níveis de até 2,0% na ração de poedeiras comerciais de ovos vermelhos não influencia o desempenho e a inclusão de 1,94% intensifica a coloração da gema.(AU)

This paper had the aim of evaluating the effect of adding annatto seeds ground in the feeds on the performance and quality of the eggs from red-egg laying hens. The treatments consisted of five diets, four experimental ones containing the inclusion of ground annatto seeds (0.5, 1.0, 1.5 and 2.0%) and a control group. A completely randomized design was adopted, with four replicates and eight birds per experimental unit. The variables analyzed were feed intake, egg production, egg mass, feed conversion (kg/kg and kg/dozen), average egg weight, Haugh unit, yolk index, color, specific gravity, pH yolk and albumen, shell percentage and thickness. The performance variables and egg quality were not affected (P>0.05) by the levels of annatto seed studied, except the color of the yolk, which presented a quadratic effect (Y=5.93+5,86x-1,51x2) demonstrating that the staining of the raw egg yolk was enhanced with the addition of 1.94% annatto seed to the feeds. It can be concluded that the addition of ground annatto seed at levels up to 2.0% in the feed of red-egg laying hens does not influence the performance, and the inclusion of 1.94% intensifies the color of the yolk.(AU)

Este estudio tuvo como objetivo evaluar el efecto de la adición de niveles de semillas de achiote molidas en piensos, sobre el rendimiento y la calidad de los huevos de gallinas ponedoras de huevos rojos. Los tratamientos se han constituido por cinco dietas, siendo cuatro experimentales conteniendo niveles de inclusión de semillas de achiote molidas (0,5, 1,0, 1,5 y 2,0%) y un grupo control. Se adoptó un diseño completamente al azar, con cuatro repeticiones y ocho aves por unidad experimental. Las variables analizadas fueron: consumo de alimento, producción de huevos, masa de huevos, conversión alimenticia (kg/kg y kg/dz), peso promedio del huevo, la unidad Haugh, índice de la yema, color, densidad, pH de yema y de albumen y grosor de la cáscara. Las variables de rendimiento y calidad de los huevos no se vieron afectados (P>0,05) por los niveles de semillas de achiote estudiados, excepto el color de la yema que mostró un efecto cuadrático (Y=5,93+5,86x-1,51x2), lo que demuestra que la tinción de la yema cruda fue intensificada con la adición de 1,94% de semillas de achiote a los piensos. Se concluye que la adición de semillas de achiote molidas a niveles de hasta 2,0% en la dieta de las gallinas ponedoras de huevos rojos no influye en el rendimiento y la inclusión de 1,94% intensifica el color de la yema.(AU)

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).

In vitro antioxidant activity of Bixa orellana (Annatto) Seed Extract.

The seeds of Bixa orellana (Annatto, family Bixaceae), have been used in food coloring for over 50 years. With the aim of introducing its extracts as pharmaceutical colorant, there is the need to investigate the biological and pharmacological activities of the extract. This study was designed to develop extraction protocols for annatto coloring fraction with potential for pharmaceutical application and evaluate the antioxidant activity of the extracts in vitro. Powdered seed material was extracted using acid-base protocols and the crystals obtained were washed with deionized water, oven-dried for about 12 hours at 45 °C and stored in air-tight containers. The in vitro antioxidant activity was tested via 2,2-diphenyl-1- picrylhydrazyl (DPPH) radical scavenging activity and iron (III) oxide reducing power using ascorbic acid (vitamin C) as a reference standard. The free radical scavenging activity of annatto extract ranged from 5.5 % to 48.9 % relative to ascorbic acid (2.9 % to 41.5 %) at respective concentrations between 0.25 and 2.5 μg/ml. Similarly, iron (III) oxide reducing power shows good linear concentration-dependent relation (R2 = 0.9986) comparable with ascorbic acid (R2 = 0.9934). Results generally indicated that Bixa orellana seed extract is a potential source of antioxidants of natural origin.

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.