Compuestos fenólicos, actividad antioxidante : bases farmacológicas y su aplicación en medicina general y oral

Los flavonoides, compuestos fenólicos, son los pigmentos responsables de la coloración de las flores, frutos y hojas, presentes en las uvas y en otros frutos como las moras, frambuesas, arándanos entre otros. Tienen actividad antioxidante comprobada. El daño tisular oxidativo y la apoptosis, pueden ser atenuados por las antocianinas, un subgrupo de los flavonoides, en células aisladas cultivadas de tejidos orales humanos. En este trabajo se describen las bases farmacológicas para la aplicación de las mismas en el tratamiento de las patologías orales donde está comprobado el stress oxidativo como vía patogénica de las mismas.

Optimización de parámetros fermentativos de calidad en vinos tintos de zonas cálidas

El aumento progresivo de la temperatura media anual y el déficit hídrico están provocando importantes cambios en la composición y la maduración de la uva, que repercuten directamente sobre el proceso fermentativo y, por ende, sobre la calidad del vino elaborado. En este trabajo se evalúan diferentes estrategias para la reducción del grado alcohólico, la mejora del color del vino y su estabilidad, y el incremento y la persistencia aromática. Mediante el empleo de levaduras con ineficiencia glicolítica se lograron reducciones medias en el grado alcohólico de entre 0.3 y 1.7 % v/v, mientras que con las fermentaciones secuenciales la máxima reducción lograda fue de 3.3 y 3.4 % v/v al combinar las cepas 938 (Schizosaccharomyces pombe) y 7013 (Torulaspora delbrueckii) con la 7VA (Saccharomyces cerevisiae). Al aplicar un tratamiento térmico sobre el inóculo, la TP2A(16) mostró una reducción media significativa en el grado alcohólico de 1 % v/v. El principal inconveniente en todas las técnicas empleadas para reducir el grado alcohólico fue la falta de repetibilidad en los resultados obtenidos. Por otra parte, la aplicación de altas presiones sobre uva despalillada resultó efectiva como tratamiento de pasteurización y como potenciador de la extracción de polifenoles, logrando un incremento en el contenido medio de antocianos totales del 12.4-18.5 %. La adición de flavonoides al mosto estimuló la formación de pigmentos estables como resultado de su condensación con antocianos mediada por acetaldehído. Con el empleo de Torulaspora delbrueckii en fermentación secuencial fue posible incrementar la producción de diacetilo y acetato de 2-feniletilo, además de la síntesis de un nuevo compuesto, el 3-etoxi-1-propanol. Sin embargo, su aportación sobre el color fue nula, así que debería combinarse con una cepa de Saccharomyces cerevisiae con buena formación de pigmentos piranoantociánicos. El empleo de Schizosaccharomyces pombe (938, V1) y Torulaspora delbrueckii (1880) en fermentaciones secuenciales y mixtas con Saccharomyces cerevisiae permitió mejorar el perfil sensorial del vino tinto mediante la mayor síntesis de polioles y la potenciación de aromas frutales, florales y herbáceos, e incrementar la estabilidad de la materia colorante al favorecer la formación de vitisinas y piranoantocianos vinilfenólicos. La crianza sobre lías en barrica a partir de levaduras seleccionadas, puede mejorar la complejidad y persistencia aromática del vino tinto, aunque sin grandes cambios en el color. ABSTRACT The progressive increase in annual average temperature, along with water deficit, is causing significant changes in grape composition and in its maturation, which directly affects the fermentative process and hence alters wine quality. In this work, different strategies for reducing the alcoholic strength, improve wine color and its stability, and increase aromatic complexity and its persistence, are evaluated. By using yeasts with glycolytic inefficiency, it was possible to achieve mean reductions between 0.3 and 1.7 % v/v in the alcoholic strength, while sequential fermentations allowed a maximum reduction of 3.3 and 3.4 % v/v by combining strains 938 (Schizosaccharomyces pombe) and 7013 (Torulaspora delbrueckii) with 7VA (Saccharomyces cerevisiae). When applying a heat shock treatment on the inoculum, only TP2A(16) strain showed a significant mean reduction of 1 % v/v in the alcohol content, compared with the control. The main drawback in all the techniques used to reduce the alcohol content was the lack of repeatability in the results. Moreover, the application of high pressures on destemmed grapes was effective as pasteurization treatment and also as enhancer of polyphenol extraction, achieving an increase of 12.4-18.5% in the average content of total anthocyanins. As expected, addition of flavonoids to the must, stimulated the formation of stable pigments, mainly as a result of condensation reactions between anthocyanins and flavanols mediated by acetaldehyde. With the use of Torulaspora delbrueckii strains in sequential fermentation with Saccharomyces cerevisiae, it was possible to increase the production of diacetyl and 2-phenylethyl acetate, besides the synthesis of a new compound: 3-ethoxy-1-propanol. The use of Schizosaccharomyces pombe (938, V1) and Torulaspora delbrueckii (1880) strains in sequential and mixed fermentations with Saccharomyces cerevisiae improved the sensory profile of red wine by increasing polyols synthesis and enhancing fruity, floral and herbaceous aromas, and it also increased the stability of the coloring matter by favouring vitisins and vinylphenolic pyranoanthocyanins formation. Ageing on lees in barrels from selected yeasts can improve the complexity and aromatic persistence of red wine, without major changes in the color.

Análise de flavanóides por cromatografia líquida de alta eficiência e eletroforese capilar - otimização de separação e aplicações tecnológicas

No presente trabalho foram estudadas as separações de 18 flavonóides (9 agliconas e 9 glicosídeos) pelas técnicas de Cromatografia Líquida de Alta Eficiência em fase reversa (RP-HPLC) e Cromatografia Micelar Eletrocinética em fluxo reverso (RF-Meck). Em ambas as técnicas foram avaliados solventes puros (metanol, acetonitrila e tetrahidrofurano) e suas misturas como formas de promover a variação de seletividade, através da modificação da fase móvel em HPLC, e da natureza do aditivo orgânico em RF-Meck. Nos estudos efetuados em HPLC utilizando-se gradiente, pode-se comprovar a possibilidade da modelagem do fator de retenção em funçã da proporção de solvente utilizados (MeOH, ACN, THF e suas misturas). Pode-se ainda, com base nos dados de retenção e na análise hierárquica de c1usters, diferenciar quatro diferentes grupos de sistemas cromatográficos com diferentes seletividades para flavonóides agliconas, e outros quatro com diferentes seletividades para glicosídeos. Os sistemas cromatográficos mais ortogonais (cada um pertencente a um grupo de seletividade) foram aplicados na separação de uma planta modelo (Azadirachta indica), de onde pode-se escolher a fase móvel mais seletiva para se otimizar a separação dos flavonóides glicosilados presentes nas folhas desta planta. No método final otimizado pode-se identificar e quantificar cinco dos flavonóides majoritários presentes, sendo três glicosídeos de quercetina (rutina, isoquercitrina e quercitrina) e dois glicosídeos de kaempferol (astragalin e nicotiflorin), em amostras de duas diferentes procedências (Piracicaba-SP e Silvânia-GO). Nos estudos envolvendo a separação dos dezoito flavonóides por RFMEKC pode-se comprovar diferenças significativas de seletividade quando se varia a natureza do solvente orgânico utilizado como aditivo, além de se observar tendências na migração em função das propriedades do solvente adicionado e da estrutura molecular do flavonóide. O solvente de menor eficiência para separação dos flavonóides foi o MeOH. Através da análise dos eletroferogramas obtidos através de um planejamento experimental de misturas, e das trocas de pares críticos observadas nos vários eletrólitos utilizados, obteve-se um método de separação com apenas um par crítico em menos de 12 minutos de corrida. O coeficiente de variação obtido para o fator de retenção foi de 1,5% e para área de 3%, considerando-se cinco injeções. O método desenvolvido foi aplicado com sucesso na identificação dos flavonóides majoritários presentes na planta modelo (Neem), obtendo-se o mesmo resultado do estudo anterior. Como forma de avaliar a concentração de flavonóides totais presentes em espécies vegetais é comum a análise de extratos após hidrólise ácida (conversão de todos glicosídeos em agliconas). Desta forma otimizou-se uma metodologia de separação em RP-HPLC de 8 flavonóides agliconas comumente presentes em alimentos e extratos vegetais de uso cosmético. A otimização foi efetuada mediante um planejamento experimental de misturas, para escolha da fase móvel mais seletiva, e de um planejamento fatorial composto central, para otimização das condições de gradiente. O método obtido foi o mais rápido já visto dentro da literatura consultada. A separação em linha de base foi efetuada em menos de 15 minutos, com coeficientes de variação de área entre 0,1 e 1,8%, coeficiente de correlação de 0,9993 a 0,9994 na faixa de 5 a 100 µg/mL, e limites de quantificação estimados na faixa de 0,1 a 0,21µg/mL. O método desenvolvido foi aplicado na otimização das condições de hidrólise de um extrato de Neem. A otimização foi efetuada através de metodologia de superfície de resposta, levando-se em consideração a concentração de ácido adicionada, o tempo de reação, a temperatura, e a concentração de um antioxidante (ácido ascórbico) adicionado. O resultado da otimização foi uma metodologia de hidrólise com tempo de reação igual a 5 minutos, utilizando-se 1,4 mol/L de HCI, 119°C e 500 µg/mL de ácido ascórbico. Através das metodologias de análise e de hidrólise desenvolvidas pode-se constatar a presença e quantificar no extrato de Neem os flavonóides agliconas quercetina, kaempferol e miricetina. Com o objetivo de se avaliar quais os componentes presentes em extratos vegetais são os responsáveis pelo poder antioxidante atribuído a determinadas plantas, foi montado um sistema de avaliação de poder antioxidante \"on-line\" com reação pós-coluna em HPLC (baseado na literatura) utilizando-se como \"radical livre modelo\" o ABTS. A análise da planta modelo (Neem) neste sistema mostrou que os flavonóides glicosilados identificados nas partes anteriores deste trabalho são os responsáveis pelo poder antioxidante atribuído a esta planta. De posse desta informação, e visando a obtenção de extratos para aplicações cosméticas com poder antioxidante, modelou-se a extração dos flavonóide do Neem em função da composição do solvente extrator (água, etanol , propilenoglicol e suas misturas), de acordo com um planejamento simplex centróide ampliado. Além da previsão da concentração dos princípios ativos pode-se ainda prever outras propriedades dos extratos obtidos, tais como, índice de refração e densidade, muitas vezes constituintes de especificações técnicas de acordo com as aplicações a que se destinam (cremes, xampús, etc).

Potencial dos biflavonóides de Araucaria angustifolia (Bert.) O. Kuntze como antioxidantes e fotoprotetores

A Araucaria angustifolia é uma conífera endêmica das regiões sul e sudeste do Brasil sendo considerada uma espécie em extinção devido ao extenso extrativismo madeireiro. Atualmente, existem inúmeros projetos visando o reflorestamento e o uso sustentável deste pinheiro. Em vista destes pontos, o estudo das propriedades dos componentes das folhas com o intuito da utilização destes com fins comerciais tornou-se de extrema importância. As suas folhas foram submetidas à extração com solventes e foram identificados seis biflavonóides majoritários, dentre estes a amentoflavona e a ginkgetina, que são apontados como agentes contra inflamações e artrites. A fração rica de biflavonóides (BFF) extraída da araucaria foi testada frente a sua atividade em proteger contra danos em biomoléculas provocadas por espécies reativas de oxigênio, capacidade em quelar metais e proteção contra raios UV. A capacidade do BFF em proteger contra danos provocados por espécies reativas de oxigênio foi comparado com compostos conhecidamente antioxidantes, como o α-tocoferol, Trolox®, quercetina, rutina e com padrões de biflavonóides, a amentoflavona e ginkgetina. O BFF demonstrou que possui uma constante de supressão do 1O2 (50 x 106 M-1s-1), superior ao da quercetina (9 x 106 M-1s-1) e foi o mais eficiente na proteção contra quebras de simples fita em DNA plasmidial, provocado por esta espécie reativa. Ainda em relação à proteção de DNA plasmidial o BFF foi capaz de proteger também contra estes danos provocados através da reação de Fenton, apesar de não demonstrar a mesma eficiência da quercetina que mostrou ser um potente protetor destes danos. O BFF protegeu contra lipoperoxidação em lipossomos de fosfatidilcolina induzida por raios UV e reação de Fenton. Em análises realizadas com espectrometria de massas foi observada a formação de complexos destes biflavonóides com íons metálicos como ferro, cobre e alumínio que possuem um papel importante na formação de radicais livres. Em relação à capacidade fotoprotetora do BFF, este inibiu a formação de dímeros de pirimidina que são apontados como causadores de câncer de pele induzidos, principalmente por radiação UV-B. Esta ação protetora foi superior àquela conferida ao p-metoxicinamato de octila, um conhecido fotoprotetor. Com o intuito de permitir a solubilização do BFF em soluções aquosas e assim, avaliar a ação do BFF em células, incorporou-se o BFF em ciclodextrina. Essa inclusão favoreceu a incorporação de BFF em células CV1-P na concentração aproximada de 0,4 µg/ml após 24 horas de incubação. Essa concentração incorporada não demonstrou ser tóxica para as células no teste com MTT. Assim, o BFF tem despertado grande interesse em relação ao seu potencial na utilização nas mais variadas áreas como cosmética, alimentos e fitoterápicos.

Obtenção de extratos da Opuntia fícus-indica L. Mill e suas aplicações em formulações cosméticas: avaliação in vivo do sensorial e da eficácia hidratante

Opuntia fícus - indica (L.) Mill is a cactacea presents in the Caatinga ecosystem and shows in its chemical c omposition flavonoids, galacturonic acid and sugars. Different hydroglicolic (EHG001 and EHG002) and hydroethanolic subsequently lyophilized (EHE001 and EHE002) extracts were developed. The EHE002 had his preliminary phytochemical composition investigated by thin layer chromatography (TLC) and we observed the predominance of flavonoids. Different formulations were prepared as emulsions with Sodium Polyacrylate (and) Hydrogenated Polydecene (and) Trideceth - 6 (Rapithix® A60), and Polyacrylamide (and) C13 - 14 I soparaffin (and) Laureth - 7 (Sepigel® 305), and gel with Sodium Polyacrylate (Rapithix® A100). The sensorial evaluation was conducted by check - all - that - apply method. There were no significant differences between the scores assigned to the formulations, howe ver, we noted a preference for those formulated with 1,5% of Rapithix® A100 and 3,0% of Sepigel® 305. These and the formulation with 3% Rapithix® A60 were tested for preliminary and accelerated stability. In accelerated stability study, samples were stored at different temperatures for 90 days. Organoleptic characteristics, the pH values and rheological behavior were assessed. T he emulsions formulated with 3,0% of Sepigel® 305 and 1,5% of Rapithix® A60 w ere stable with pseudoplastic and thixotropic behavior . The moisturizing clinical efficacy of the emulsions containing 3,0% of Sepigel® 305 containing 1 and 3% of EHG001 was performed using the capacitance method (Corneometer®) and transepidermal water lost – TEWL evaluation ( Tewameter®). The results showed t hat the formulation with 3% of EHG001 increased the skin moisturizing against the vehicle and the extractor solvent formulation after five hours. The formulations containing 1 and 3% of EHG001 increased skin barrier effect by reducing transepidermal water loss up to four hours after application.

Extração do corante do fruto de castanhola (Terminalia catappa Linn) e estudos dos seus compostos fenólicos, antocianinas e atividade antioxidante

The Terminalia catappa Linn belonging to Combretaceae family, popularly known as castanets, has fruits consists of a fleshy pulp, rounded seed and a very hard shell. The natural pigmentation existing in the fruit of castanet indicates the presence of anthocyanins, phenolic nature components belonging to the group of flavonoids, which have antioxidant activity. This research was conducted with the castanets and aimed to the study of factors influencing the extraction of dyes from its pulp. The extracts were obtained using a reactor enjaquetado by solid-liquid extraction. The factors were evaluated as temperature, time, solvent ratio and pH extraction. Adopting a factorial design of 24 , with 4 repetitions at the central point, the effects of these factors on the extraction process were analyzed using Statistica 7.0 software. The antioxidant activity (AA), the content of phenolic compounds (CFT) and the total monomeric anthocyanin content (AMT) were evaluated as response variables planning. Statistical analysis of the results, the effects that influenced the extraction were different for each response (CFT, AMT and AA). However, the pH was significant for the extraction of all compounds. The kinetic behavior of the dye extraction was also studied for phenolic compounds, monomeric anthocyanins and antioxidant activity, in which the equilibrium was reached after 90 minutes of extraction. To study the stability of anthocyanins temperature was the factor that most influenced the stability, however the concentration and pH also played a part.

Avaliação anti-inflamatória e antipeçonhenta das espécies Hancornia speciosa e Mimosa tenuiflora em modelos experimentais de inflamação e envenenamento induzidos por Bothrops jararaca e Tityus serrulatus

Accidents caused by venomous animals represents a significant and serious public health problem in certain regions of Brazil, as well as in other parts of the world by the frequency with which they occur and the mortality they cause. The use of plant extracts as an antidote for poisoning cases is an ancient practice used in many communities that have no access to antivenom. Medicinal plants represent an important source of obtaining bioactive compounds able to assist directly in the treatment of poisoning or indirectly supplementing serum therapy currently used. The aim of this study was to evaluate the effect of extracts, fractions and isolated compounds from M. tenuiflora and H. speciosa in the inflammatory process induced by carrageenan and the venom of B. jararaca and T. serrulatus. The results showed that both M. tenuiflora and H. speciosa were capable of inhibiting cell migration and cytokines levels in peritonitis induced by carrageenin and venom of T. serrulatus. In poisoning by B. jararaca model, mice treated with the plants in studies decreased the leukocyte influx into the peritoneal cavity. Finally the M. tenuiflora and H. speciosa had antiphlogistic activity, reducing edema formation and exerted inhibitory action of leukocyte migration in local inflammation induced by the venom of B. jararaca. Through of Thin Layer Chromatography (TLC) analysis was possible identified the presence of flavonoids ,saponins and/or terpenes in aqueous extract of M. tenuiflora. By High Performance Liquid Chromatography analysis, it was possible to identify the presence of rutin and chlorogenic acid in aqueous extract of H. speciosa. We conclude that the administration of extracts, fractions and isolated compounds of H. speciosa and M. tenuiflora resulted in inhibition of the inflammatory process in different experimental models. This study demonstrates for the first time the effect of M. tenuiflora and H. speciosa in inhibition of the inflammation caused by B. jararaca and T. serrulatus venom.

Spondias tuberosa Arr. (UMBU): estudo fitoquímico e avaliação do potencial anti-inflamatório

Spondias tuberosa Arruda (Anacardiaceae) is a fruitful tree popularly known as umbuzeiro, tapereba or umbu. It is a native and endemic species from Brazil, widespread in Brazilian Northeast. The species is important in folk medicine of the semi-arid Northeast, where it is mainly used to treat various inflammatory conditions, digestive problems as well as viral and bacterial infections. However, despite the common use in folk medicine, there are scarce pharmacological and phytochemicals studies that afford scientific evidence to its popular use. Therefore, this study aimed to characterize the chemical markers in S. tuberosa leaves extract, obtained by maceration ethanol:water (70:30, [v/v]), and evaluate its anti-inflammatory potential in vivo. The phytochemical profile in TLC analysis suggested the occurence of the flavonoids rutin and isoquercitrin. HPLC analysis enabled us to confirm the presence of flavonoids and also, were detected the phenolic acids, chlorogenic acid and caffeic acid. In addition was developed and validated a HPLC method to evaluate the content of the identified compounds in S. tuberosa leaves extract according to RDC 899/2003 of ANVISA and ICH Guidelines 2005. In order to evaluate the anti-inflammatory potential of S. tuberosa leaves extract, the peritonitis and paw edema models induced by carrageenan were used, administration i.p. in mice. The results highlighted the anti-inflammatory property in vivo at 125, 250 and 500 mg/kg since a decrease in leukocyte influx to the site of inflammation, diameter of the edema and the level of myeloperoxidase were observed when compared to the drug control dexamethasone (2 mg/kg, i.p. route). Taken together, the results pointed out S. tuberosa as a potential species for developing phytotherapic derivatives in according to its popular use. With regard to the characterization markers, chlorogenic acid, caffeic acid, rutin and isoquercitrin were identified and quantified in Spondias tuberosa leaves extract so they could be used in quality control analyses of the raw material and extracts of this species.

Avaliação da toxicidade subcrônica e reprodutiva do extrato seco de pericarpo de passiflora edulis variedade flavicarpa degener em ratos

The pericarp of Passiflora edulis var. flavicarpa Degener is now being investigated for medicine purposes. There are no reports about it toxicity. The aim of the present study was investigate the sub chronic toxicity in male rats and reproductive toxicity in pregnant rats and exposed fetuses of an extract obtained by infusion of the pericarp in water (1:3 m/v;100o C, 10 min). The extract composition was evaluated by tube reactions and thin lawyer chromatography (TLC). Adult male rats (n=8) were treated with 300 mg/kg of the extract, by gavage, during 30 days and pregnant rats (n=7) from gestation day 0 to day 20. Control received tap water (1 mL). Water and food intakes and body weight gain were recorded. At day 29 of treatment the sexual behavior of the males was analyzed and then half of males from each group received cyclophosphamide (50 mg/kg, i.p.) to (anti)genotoxic assessment in bone marrow. At day 30, males were anesthesized for parameters collection. At day 20 of gestation, the dams were anesthesized for reproductive performance evaluation. The fetal analysis was conducted by visceral and skeletal. Phytochemical analysis revealed the presence of flavonoids, unspecific alkaloids, phenols and triterpenic compounds. Statistical analysis revealed absence of significant differences between experimental and control. This study suggest that the aqueous extract obtained from pericarp of P. edulis var. flavicarpa Degener was not able to promote toxic effects in rats. Cytotoxicity was evaluated with the PCE/NCE ratio (NCE=normochromatic erythrocytes). Statistical analysis (mean ± SEM) revealed absence of changes in the frequency of MNPCE (negative control: 3.26±0.42; positive control: 11.72±1.02; negative experimental: 4.02±0.13; positive experimental: 10.47±0.87) or cytotoxicity (negative control: 0.37±0.08; positive control: 0.23±0.05; negative experimental: 0.37±0.07; positive experimental: 0.23±0.02). This study suggests that the extracts showed no (anti)genotoxic and no cytotoxic activities under the experimental conditions.

Estudo fitoquímico de Waltheria ferruginea

The Waltheria genus belonging to the Sterculiaceae family, it is reported as a prolific source of flavonoids and quinolone alkaloids, substances of great interest due to several associated biological activities. This work describes a novel phytochemical study from Waltheria ferruginea, aiming to contribute to the chemical knowledge of this specie and the isolation of substances with biological potential. For the phytochemical study were used chromatography techniques on silica gel and molecular exclusion in Sephadex LH-20.The structural elucidation of the isolated compounds was performed through spectrometric techniques 1H and 13C NMR, including uni and bidimensional pulse sequences, and comparison with data from literature. Five substances were isolated, namely: the flavonids kaempferol-3-O-β-(6''-cumaroil)-glucopyranoside (F1) and kaempferol -3 -O- β - glucopyranoside (F2), both analyzes with pharmacological properties, the flavonol quercetin-3-O-β-glucopyranoside (F3 ) pure and in the epimeric mixture α (F3') and (F3), the terpenegeranyl - geranyl (G1) and the 12-hydroxi-octadecanoic acid, all no previous reported in the literature.

Inibição dos efeitos locais induzidos pelas peçonhas das serpentes Bothrops erythromelas e Bothrops jararaca pelo extrato aquoso das folhas de Jatropha mollissima (Pohl) Bail

Snakebites are a serious public health problem in tropical and subtropical countries and Bothrops genus is responsible for the accidents in Brazil and throughout Latin America (90% of cases). The local effects (pain, edema, hemorrhage and myonecrosis) and systemic (cardiovascular alterations, shock and blood clotting disorders) caused by the venom of Bothrops are due to the numerous protein and non-protein components, which are part of the constitution of the poison. The only form of therapy is scientifically validated antivenom serum therapy which, however, is not effective with respect to local effects produced, risk of immunological reactions, high cost and difficult access in some regions. Thus, the search for new alternatives to serum therapy becomes important, and in this context, many medicinal plants have been highlighted by the popular use as antiophidic. Among these plants, we can mention the species Jatropha mollissima (Euphorbiaceae) which has popular use in traditional medicine as antiophidic, anti-inflammatory, antimicrobial and antipyretic. Therefore, this study aims to evaluate the neutralizing potential of local effects induced by the venom of Bothrops erythromelas and Bothrops jararaca with the aqueous extract of the leaves of J. mollissima. The leaf extracts were prepared by decoction, fractionated (by liquid-liquid partition) and characterized by thin layer chromatography (TLC) and High Performance Liquid Chromatography (HPLC). Antiophidic activity of the extract was evaluated in model of paw edema, peritonitis, bleeding and myotoxicity induced by venoms of B. jararaca and B. erythromelas. In all models, the extract was evaluated by intraperitoneal route at the doses of 50, 100 and 200 mg/kg, administered 30 minutes prior to injection of the venom (pretreatment protocol). Stains suggestive of the presence of flavonoids: apigenin, luteolin, orientin, isoorientin, vitexin and vitexin-2-O-rhamnoside were detected in the extract by co-CCD. By means of HPLC were identified isoorientin, orientin, vitexin and isovitexin. All tested doses of J. mollissima extract reduced the paw edema induced by the venom with intensity similar to dexamethasone. The aqueous extract of J. mollissima leaves on all evaluated doses, inhibited cell migration induced by B. jararaca and B. erythromelas promoting inhibition of recruitment of mononuclear cells and the polymorphonuclear cells. Local bleeding induced by B. jararaca venom was significantly inhibited by the extract. Both venoms were inhibited by the extract in myotoxic activity. These results indicate that the aqueous extract of J. mollissima leaves have snakebite potential, particularly with respect to local effects, which may justify the use of this plant in traditional medicine and complementary therapy as anti-venom serum.

Inibição dos efeitos locais induzidos pelas peçonhas das serpentes Bothrops erythromelas e Bothrops jararaca pelo extrato aquoso das folhas de Jatropha mollissima (Pohl) Bail

Snakebites are a serious public health problem in tropical and subtropical countries and Bothrops genus is responsible for the accidents in Brazil and throughout Latin America (90% of cases). The local effects (pain, edema, hemorrhage and myonecrosis) and systemic (cardiovascular alterations, shock and blood clotting disorders) caused by the venom of Bothrops are due to the numerous protein and non-protein components, which are part of the constitution of the poison. The only form of therapy is scientifically validated antivenom serum therapy which, however, is not effective with respect to local effects produced, risk of immunological reactions, high cost and difficult access in some regions. Thus, the search for new alternatives to serum therapy becomes important, and in this context, many medicinal plants have been highlighted by the popular use as antiophidic. Among these plants, we can mention the species Jatropha mollissima (Euphorbiaceae) which has popular use in traditional medicine as antiophidic, anti-inflammatory, antimicrobial and antipyretic. Therefore, this study aims to evaluate the neutralizing potential of local effects induced by the venom of Bothrops erythromelas and Bothrops jararaca with the aqueous extract of the leaves of J. mollissima. The leaf extracts were prepared by decoction, fractionated (by liquid-liquid partition) and characterized by thin layer chromatography (TLC) and High Performance Liquid Chromatography (HPLC). Antiophidic activity of the extract was evaluated in model of paw edema, peritonitis, bleeding and myotoxicity induced by venoms of B. jararaca and B. erythromelas. In all models, the extract was evaluated by intraperitoneal route at the doses of 50, 100 and 200 mg/kg, administered 30 minutes prior to injection of the venom (pretreatment protocol). Stains suggestive of the presence of flavonoids: apigenin, luteolin, orientin, isoorientin, vitexin and vitexin-2-O-rhamnoside were detected in the extract by co-CCD. By means of HPLC were identified isoorientin, orientin, vitexin and isovitexin. All tested doses of J. mollissima extract reduced the paw edema induced by the venom with intensity similar to dexamethasone. The aqueous extract of J. mollissima leaves on all evaluated doses, inhibited cell migration induced by B. jararaca and B. erythromelas promoting inhibition of recruitment of mononuclear cells and the polymorphonuclear cells. Local bleeding induced by B. jararaca venom was significantly inhibited by the extract. Both venoms were inhibited by the extract in myotoxic activity. These results indicate that the aqueous extract of J. mollissima leaves have snakebite potential, particularly with respect to local effects, which may justify the use of this plant in traditional medicine and complementary therapy as anti-venom serum.

Atividade antimicrobiana e antioxidante de Licania rigida e Turnera ulmifolia

Introduction: Licania rigida Benth and Turnera ulmifolia Linn. var. elegans are species of semi-arid regional plants used in the treatment of various diseases. Objectives: The purpose of this study was chemically characterize the extracts and fractions and investigate the antimicrobial and antioxidant potential. Methods: For chemical analysis, were performed spectrophotometric quantification of the total phenolic and characterization of the extracts by chromatographic analysis. Evaluation of antioxidant activity was done by determining the radical scavenging capacity DPPH •. Antimicrobial activity was evaluated by agar diffusion, broth microdilutionand time-kill assays. Results: The extracts and fractions L. rigid and T. ulmifolia showed a high phenolic content, the presence of flavonoids, which were determined as chemical markers. It was observed that the extracts of both species performed as sequestering agents in the trial of antioxidant activity in vitro. The L. rigida extract was the only active front strains of S. aureus 33591 (methicillin-resistant), S. aureus 29213, S. epidermidis 12228, and also against the yeast, Candida albicans, Candida dubliniensis, Candida tropicalis, Candida parapsilosis, Candida rugosa, Candida krusei eTrichosporon asahii. Conclusions: Based on these results it is possibly affirm the antioxidant and antimicrobial activity of L. rigida and attributed the presence of polyphenolic flavonoid like responsible.

Prospecção química e avaliação biológica de Banisteriopsis laevifolia (Malpighiaceae)

The Banisteriopsis genus is widespread in traditional medicine. This work aims to contribute with information about the chemical composition and on the evaluation of the biological activity of the essential oil, the ethanol extract of the leaves and partitions of the Banisteriopsis laevifolia. The phytochemical screeningtest of ethanol extract and partitions of leaves indicated the presence of flavonoids, terpenoids, saponins, phenols and steroids compounds. Nitrogenous compounds, characteristic of some species of this family, were not detected. Flavonoids were the predominant metabolite, with the highest concentrations on the partitions ethyl acetate and n-butanol. The antibacterial activity, antifungal and cytotoxicity of the essetial oil, ethanol extract and partitions were assyed by microdilution broth method (MBM), where the minimum inhibitory concentrations (MIC) were calculated. The ethanol extract and partitions did not inhibit growth against to Gram positive bacteria tested, with MIC less than 400 mg L-1. For the Gram negative bacteria tested, the hexane and hydroethanol partitios were more effective against F. nucleatum bacteria (MIC 100 ug mL-1). The ethanol extract showed antifungal activity with MIC of 31.2 mg L-1. Ethyl acetate and n-butanol partitions showed MIC 187.5 mg L-1 and 93.7 mg L-1, respectively, arousing interest for isolation studies. The antioxidant activity was evaluated by the DPPH free radical method. The ethanolic extract, ethyl acetate and n-butanol partitions were active, since they showed EC50 values (4.53 ug mL-1, 4.07 and 8.39 ug mL-1, respectively), values equivalent to the BHT (7.3 mg L-1). The analysis by HPLC-MS/MS of the most active fractions (ethyl acetate and n-butanol) identified phenolic compounds (flavonols and phenolic acids) which exert recognized biological activity. The GC-MS analysis of the essential oils from leaves collected in two periods studied (dry and wet), showed a small variation in the number of compounds. The major classes identified for the oil collected in the dry period were aliphatic alcohols (23,4%), terpenoids (18.7%), sterols (10.4%) and long-chain alkanes (9.2%) compounds. Terpenoids (26.8%) were the major class for the rain season. The major compounds (3Z) -hexenol, phytol and untriacontano are present in the two seasons but in different amounts (19.4%, 9.8% and 7.5% during the dry season, and 17.0 %, 14.9% and 15.3% in the rainy season, respectively). The essential oil from rainy season was not effective against to the oral bacteria Gram positive and Gram negative tested. However, showed significant antifungal activity with MIC 1000 mg L-1 against Candidas. Thus, the promising results with respect to biological assays of ethanolic extract and partitions from B. laevifolia contributed to the chemical and biological knowledge of the species B. laevifolia.

Estudo fitoquímico e biológico das folhas de Banisteriopsis argyrophylla (A. Juss.) B. Gates (Malpighiaceae)

With the increasing fungi resistance compared with existing drugs on the market and the side effects reported by some compounds with antioxidant properties and enzymatic inhibitors, in particular against α-amylase and α-glucosidase, the discovery of new compounds with biological potential, becomes a need. In this context, natural products can be an important source for the discovery of new active molecular architectures. Then, this study aimed to evaluate the antioxidant activity, the enzymatic inhibitory activity of α-amylase and α-glucosidase, the antifungal and cytotoxic activities of ethanolic extract (EE) the leaves of Banisteriopsis argyrophylla (Malpighiaceae) and their fractions, obtained by liquid-liquid extraction using solvents of increasing polarity. The antioxidant activity was evaluated by the free radical DPPH scavenging method (2,2-diphenyl-1-picrylhydrazyl) and the ethyl acetate fractions (FAE) and n-butanol (FB) were the most active, confirmed by the peak current and the oxidation potential obtained by differential pulse voltammetry (DPV). The inhibitory activity of the α-amylase and α-glucosidase was analyzed considering the reactions between substrates α-(2-chloro-4-nitrophenyl)-β-1,4-galactopiranosilmaltoside (Gal-α-G2-CNP) and 4-nitrophenyl-α-D-glucopyranoside (p-NPG), respectively. Initially, it was found that the EE showed considerable activity against α-amylase (EC50 = 2.89±0.1 μg m L–1) compared to the acarbose used as positive control (EC50 = 0.08±0.1 μg mL–1) and that did not showed promising activity against the α-glucosidase. After this observation we evaluated the inhibitory activity of α-amylase fractions, with FAE (EC50 = 2.33±0.1 μg mL–1) and FB (EC50 = 2.57 ± 0.1 μg mL–1) showing the best inhibitions. The antifungal activity was evaluated against Candida species, and the FAE had better antifungal potential (MIC's between 93.75 and 11.72 μg mL–1) compared with amphotericin as positive standard (MIC = 1.00 and 2.00 μg L–1 for C. parapsilosis and C. krusei used as controls, respectively). The EE (CC50 = 360.00 ± 12 μg mL–1) and fractions (CC50's> 270.00 μg mL–1) were considerably less toxic to Vero cells than the cisplatin used as positive control (CC50 = 7.01 ± 0 6 μg mL–1). The FAE showed the best results for the activities studied, this fraction was submitted to ultra performance liquid chromatography coupled with mass spectrometry (UPLC-MS)), and the following flavonoids have been identified: (±)-catechin, quercetin-3-O-β-D-Glc/ quercetin-3-O-β-D-Gal, quercetin-3-O-β-L-Ara, quercetin-3-O-β-D-Xyl, quercetin-3-O-α-L-Rha, kaempferol-3-O-α-L-Rha, quercetin-3-O-(2''-galoil)-α-L-Rha, quercetin-3-O-(3''-galoil)-α-L-Rha and kaempferol-3-O-(3''-galoil)-α-L-Rha,. FAE was submitted to column chromatography using C18 phase, and (±)-catechin was isolated (FAE-A1, 73 mg) and three fractions consisting of a mixture of flavonoids were obtained (FAE-A2, FAE-A3 and FAE-A4). These compounds were identified by thin layer chromatography (TLC) and (–)-ESI-MS. The (±)-catechin fraction showed an MIC = 2.83 μg ml–1 in assay using C. glabrata, with amphotericin as positive control. The fractions FAE-A2, FAE-A3, FAE-A4, showed less antifungal potential in tested concentrations. The identified flavonoids are described in the literature, regarding their antioxidant capacity and (±)-catechin, quercetin-3-O-Rha and kaempferol-3-O-Rha are described as α-amylase inhibitors. Thus, B. argyrophylla is an important species that produces compounds with antioxidant potential that can be related to the traditional use as anti-inflammatory and also has antifungal compounds and inhibitors of α-amylase. Therefore, these leaves are promising resources for the production of new drugs.