Determinação quantitativa de taninos em três espécies de Stryphnodendron por cromatografia líquida de alta eficiência

A method of separation and quantification by reverse-phase high-performance liquid chromatography (HPLC) was developed, using water (0.05% TFA):acetonitrile (0.05% TFA) as the mobile phase in a gradient system. Flavan-3-ols present in a semipurified extract from the stem bark of Stryphnodendron adstringens, Stryphnodendron polyphyllum and Stryphnodendron obovatum were analyzed. The HPLC was performed with the ethyl-acetate fraction (EAF) using a solid-phase extraction on cartridges C18-E with methanol:water (2:8), filtered through a membrane of 0.5 μm pore size; the column was Phenomenex® Gemini C-18 (5 μm) at 30 ºC, with a flow rate of 0.8 mL/min. The analysis was done at 210 nm. Gallic acid and gallocatechin solutions were used as calibration standards. The proposed method was validated by resolution RE No. 899/2003 of the National Health Surveillance Agency. Quantitative analysis of the EAF showed high contents of flavan-3-ols in the stem bark of all three species. This study demonstrated that it is possible to determine the concentration of individual substances in tannin-rich plants. The system developed can be used as a chromatographic profile for the semipurified fraction of S. adstringens, S. polyphyllum, and S. obovatum.Um método de separação e quantificação por cromatografia líquida de alta eficiência (CLAE) em fase reversa foi desenvolvido usando água (0,05% de TFA):acetonitrila (0,05% de TFA) como fase móvel, em sistema gradiente para a análise dos flavan-3-óis presentes em extrato semipurificado das cascas de Stryphnodendron adstringens, Stryphnodendron polyphyllum e Stryphnodendron obovatum. A CLAE foi realizada com a fração acetato de etila (FAE) sendo submetida à extração em fase sólida (cartucho C18-E) com metanol:água (2:8), filtrada por membrana de porosidade 0,5 μm; a pré-coluna e coluna empregadas foram Phenomenex® Gemini C-18 (5 μm), com esta última mantida a 30 ºC, com vazão de 0,8 mL/min e detecção a 210 nm. Utilizaram-se soluções dos padrões de ácido gálico e galocatequina para a obtenção da curva analítica. O método proposto foi validado de acordo com a resolução RE n° 899/2003 da ANVISA. A análise quantitativa da FAE das três espécies mostrou que existe similaridade no teor de galocatequina. S. adstringens possui ácido gálico em uma proporção superior a 60% em relação às outras duas espécies. A metodologia desenvolvida mostrou-se viável à aplicação em plantas ricas em taninos, como nos casos de S. adstringens, S. polyphyllum e S. obovatum

Chemotaxonomic significance of 5-deoxyproanthocyanidins in Stryphnodendron species

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Antifungal Properties of Crude Extracts, Fractions, and Purified Compounds from Bark of Curatella americana L. (Dilleniaceae) against Candida Species

The ethnomedicinal plant Curatella americana L. (Dilleniaceae) is a common shrub in the Brazilian cerrado, in which crude extract showed antifungal activity in a preliminary study. In this work, the antifungal and cytotoxic properties of the crude extract, fractions, and isolated compounds from C. americana were evaluated against the standard yeast strains Candida albicans, C. tropicalis, and C. parapsilosis, clinical isolates, and fluconazole-resistant strains. The combinatory effects between subfractions and isolated compounds and effects on cell morphology, virulence factors, and exogenous ergosterol were also evaluated. The MIC obtained against the Candida species including fluconazole-resistant strain ranged from 15.3 to 31.3 µg/mL for crude extract, 3.9 to 15.6 µg/mL for ethyl acetate fraction, and 7.8 to 31.3 µg/mL for subfractions. The isolated compounds identified as 4′-O-methyl-catechin, epicatechin-3-O-gallate, and 4′-O-methyl-catechin-3-O-gallate showed lower antifungal activity than the crude extract and fractions (MIC ranging from 31.3 to 125.0 µg/mL). The addition of exogenous ergosterol to yeast culture did not interfere in the antifungal activity of the extract and its fractions. Synergistic antifungal activity was observed between subfractions and isolated compounds. The effects on virulence factors and the different mechanisms of action compared to fluconazole and nystatin suggest that this ethnomedicinal plant may be an effective alternative treatment for candidiasis

Phytochemistry

Texto completo. Acesso restrito. p. 71–77The plant Cecropia pachystachya Trécul is widely used in Brazilian ethnomedicine to treat hypertension, asthma, and diabetes. Arginase is an enzyme with levels that are elevated in these disorders, and it is central to Leishmania polyamine biosynthesis. The aims of this study were to evaluate antileishmanial activity and inhibition of the arginase enzyme by C. pachystachya extracts, and to study changes in cellular organization using electron microscopy. The ethanol extract of C. pachystachya was tested on Leishmania (Leishmania) amazonensis promastigote survival/proliferation and arginase activity in vitro. Qualitative ultrastructural analysis was also used to observe changes in cell organization. The major bioactive molecules of the ethanol extract were characterized using liquid chromatography–electrospray ionizationmass spectrometry (LC–ESI-MS). The ethyl acetate fraction of the ethanol extract diminished promastigote axenic growth/survival, inhibited arginase activity, and altered a mitochondrial kinetoplast DNA (K-DNA) array. The bioactive compounds of C. pachystachya were characterized as glucoside flavonoids. Orientin (9) (luteolin-8-C-glucoside) was the main component of the methanol-soluble ethyl acetate fraction obtained from the ethanol extract and is an arginase inhibitor (IC50 15.9 lM). The ethyl acetate fraction was not cytotoxic to splenocytes at a concentration of 200 lg/mL. In conclusion, C. pachystachya contains bioactive compounds that reduce the growth of L. (L.) amazonensis promastigotes, altering mitochondrial K-DNA arrangement and inhibiting arginase.Salvado

Biological activity and quality control of extract and stem bark from Stryphnodendron adstringens

Fueron evaluadas las actividades antibacteriana e hipotensora del extracto acetona: agua y de extractos semipurificados del tallo de Stryphnodendron adstringens. El extracto crudo y los extractos semipurificados mostraron actividad contra Pseudomonas aeruginosa y Staphylococcus aureus. Sin enbargo no fue posible confirmar la actividad hipotensora de los extractos de Stryphnodendron adstringens. El control de calidad fue determinado con la droga vegetal colectada por dos años, a través de ensayos farmacopeicos y cromatógraficos.The antibacterial and hypotensive activities of an acetone:water and semipurified extracts from the stem bark of Stryphnodendron adstringens were evaluated. Both the crude and semipurified extracts showed activity against Pseudomonas aeruginosa and Staphylococcus aureus. It wasn't possible to confirm the hypotensive activity. Quality control was determined, using the vegetable drug for two years, by means of pharmacopoeial and chromatographic methods.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Leishmanicidal activity of Cecropia pachystachya flavonoids: arginase inhibition and altered mitochondrial DNA arrangement

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2013-10-23T17:55:30Z No. of bitstreams: 1 Cruz, ED Leishmanicidal activity....pdf: 753862 bytes, checksum: c99ee95789e442d656e72a0f4b9edb0c (MD5)Made available in DSpace on 2013-10-23T17:55:30Z (GMT). No. of bitstreams: 1 Cruz, ED Leishmanicidal activity....pdf: 753862 bytes, checksum: c99ee95789e442d656e72a0f4b9edb0c (MD5) Previous issue date: 2013Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Biologia Parasitária. Salvador, BA, BrasilUniversidade de São Paulo. Faculdade de Zootecnia e Engenharia de Alimentos. Departamento de Medicina Veterinária. São Paulo, SP, BRasilUniversidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Departamento de Fisiologia. Ribeirão Preto, SP, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Biologia Parasitária. Salvador, BA, BrasilUniversidade de São Paulo. Faculdade de Zootecnia e Engenharia de Alimentos. Pirassununga, SP, BrazilUniversidade de São Paulo. Faculdade de Zootecnia e Engenharia de Alimentos. Departamento de Medicina Veterinária. São Paulo, SP, BRasilCurso de Farmácia. Unidade de Ensino Superior Ingá. Maringa, SP, BrazilUniversidade Federal da Bahia. Instituto de Química. Departamento de Química Orgânica. Salvador, BA, BrazilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Biologia Parasitária. Salvador, BA, BrasilThe plant Cecropia pachystachya Trécul is widely used in Brazilian ethnomedicine to treat hypertension, asthma, and diabetes. Arginase is an enzyme with levels that are elevated in these disorders, and it is central to Leishmania polyamine biosynthesis. The aims of this study were to evaluate antileishmanial activity and inhibition of the arginase enzyme by C. pachystachya extracts, and to study changes in cellular organization using electron microscopy. The ethanol extract of C. pachystachya was tested on Leishmania (Leishmania) amazonensis promastigote survival/proliferation and arginase activity in vitro. Qualitative ultrastructural analysis was also used to observe changes in cell organization. The major bioactive molecules of the ethanol extract were characterized using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). The ethyl acetate fraction of the ethanol extract diminished promastigote axenic growth/survival, inhibited arginase activity, and altered a mitochondrial kinetoplast DNA (K-DNA) array. The bioactive compounds of C. pachystachya were characterized as glucoside flavonoids. Orientin (9) (luteolin-8-C-glucoside) was the main component of the methanol-soluble ethyl acetate fraction obtained from the ethanol extract and is an arginase inhibitor (IC50 15.9 µM). The ethyl acetate fraction was not cytotoxic to splenocytes at a concentration of 200 µg/mL. In conclusion, C. pachystachya contains bioactive compounds that reduce the growth of L. (L.) amazonensis promastigotes, altering mitochondrial K-DNA arrangement and inhibiting arginase

Biological activity and quality control of extract and stem bark from Stryphnodendron adstringens

Fueron evaluadas las actividades antibacteriana e hipotensora del extracto acetona: agua y de extractos semipurificados del tallo de Stryphnodendron adstringens. El extracto crudo y los extractos semipurificados mostraron actividad contra Pseudomonas aeruginosa y Staphylococcus aureus. Sin enbargo no fue posible confirmar la actividad hipotensora de los extractos de Stryphnodendron adstringens. El control de calidad fue determinado con la droga vegetal colectada por dos años, a través de ensayos farmacopeicos y cromatógraficos.The antibacterial and hypotensive activities of an acetone:water and semipurified extracts from the stem bark of Stryphnodendron adstringens were evaluated. Both the crude and semipurified extracts showed activity against Pseudomonas aeruginosa and Staphylococcus aureus. It wasn't possible to confirm the hypotensive activity. Quality control was determined, using the vegetable drug for two years, by means of pharmacopoeial and chromatographic methods.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Biological effects of extracts obtained from Stryphnodendron adstringens on Herpetomonas samuelpessoai

We report the effect of Stryphnodendron adstringens on the trypanosomatid Herpetomonas samuelpessoai. The parasites were grown at 28ºC in a chemically defined medium containing crude extract and fractions at concentrations from 100 to 5000 µg/ml obtained from S. adstringens. Concentrations of 500, 1000, 2500, and 5000 µg/ml both crude extract and semi-purified fraction progressively inhibited the protozoans' growth. At a concentration of 100 µg/ml, crude extract or a semi-purified (F3) fraction did not affect the growth of the protozoans. The F3-9 - F3-12 sub-fractions, at a concentration of 1000 µg/ml, also showed increased inhibitory activity on H. samuelpessoai. The IC50 of the crude extract and the F3 fraction were 538 and 634 µg/ml, respectively. Ultrastructural and enzymatic alterations in the trypanosomatids were also evaluated. H. samuelpessoai cultivated in the presence of IC50 crude extract showed considerable ultrastructural alterations, such as marked mitochondrial swelling with a large number of cristae and evident Golgi complex vesiculation, as observed by transmission electron microscopy. Cells exposed to 538 µg/ml of crude extract at 28ºC for 72 h, showed decreased activity of the enzyme succinate cytochrome c reductase, a typical mitochondrion marker, as compared to untreated cell