Estudo químico e biológico do extrato etanol 70% das folhas de Solanum paniculatum L. (Solanaceae)
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Data
2018-11-23
Autores
Valerino Diaz, Alexander Bárbaro
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Universidade Estadual Paulista (Unesp)
Resumo
A espécie Solanum paniculatum conhecida popularmente como jurubeba, é amplamente usada pela população brasileira pelos seus usos medicinais para o tratamento de disfunções digestivas e doenças do fígado. A espécie é a única dentro do gênero Solanum que está incluída na lista do Sistema Único de Saúde (SUS) de espécies para o desenvolvimento de formulações de fitoterápicos. Como parte de nossa contribuição ao entendimento em relação ao uso e benefício desta espécie, o objetivo do presente trabalho consistiu no estudo químico do extrato etanol 70 % obtido a partir das folhas de S. paniculatum, através da identificação, isolamento e elucidação estrutural dos principais metabólitos presentes no extrato, assim como avaliar as atividades citotóxica do extrato etanol 70% para linhagens tumorais, antibacteriana, leishmanicida, esquistossomosse e de viabilidade celular para células normais, tanto para o extrato como para a fração enriquecida de saponinas (f’26). Diferentes técnicas cromatográficas de isolamento e purificação (Sephadex LH-20, Cromatografia Líquida de Alta Eficiência acoplada ao Detector de Arranjo de Diodo (CLAE-DAD), Cromatografia Líquida de Alta Eficiência acoplada ao Detector de Índice de Refracção (CLAE-IR)) foram empregadas, o que possibilitou o isolamento de 11 substâncias que foram caracterizadas utilizando técnicas espectroscópicas (uni e bidimensionais) e a espectrometria de massas. O fraccionamento do extrato etanol 70 % levou ao isolamento de 8 saponinas esteroidais inéditas denominadas: 6-O-α-L-ramnopiranosil-(1’’→3’)-β-D-quinovopiranosil-(22S,23R,25S)-3β,6α,23-triidroxi-5α-espirostano (1), 6-O-α-L-ramnopiranosil-(1’’→3’)-β-D-quinovopiranosil-(22S,23R,25R)-3β,6α,23-triidroxi-5α-espirostano (2), 6-O-β-D-xilopiranosil-(1’’→3’)-β-D-quinovopiranosil-(22S,23R,25S)-3β,6α,23-triidroxi-5α-espirostano (3), 6-O-β-D-xilopiranosil-(1’’→3’)-β-D-quinovopiranosil-(22S,23R,25R)-3β,6α,23-triidroxi-5α-espirostano (4), 3-O-α-L-ramnopiranosil-(1’’→3’)-β-D-quinovopiranosil-(22S,23S,25R)-3β,6α,23-triidroxi-5α-espirostano (5), 3-O-β-D-xilopiranosil(1’’→3’)-β-D-quinovopiranosil-(22S,23S,25R)-3β,6α,23-triidroxi-5α-espirostano (6), 6-O-α-L-ramnopiranosil-(1’’→3’)-β-D-quinovopiranosil-(22S,25S)-1β,3β,6α-triidroxi-5α-espirostano (7), 6-O-β-D-xilopiranosil-(1’’→3’)-β-D-quinovopiranosil-(22S,25S)-3β,4β,6α-triidroxi-5α-espirostano (8), assim como a rutina (9), o ácido cafeico (10) e o anidrido cafeico (10a). Os teores das saponinas (1-8) no extrato etanol 70 % foram realizadas por quantificação por UPLC-ESI-IT-MS, e demostrou-se os altos conteúdos de saponinas que apresentou o extrato etanol 70 % (333,6 0,09 mg.g-1 do extrato). O ensaio de citotoxicidade do extrato etanol 70 % mostrou uma baixa toxicidade com valores de CIM50 superiores a 192,92 µg mL-1 para cada linhagem testada, no entanto, o extrato etanol 70 %, assim como a fração (f’26) mostraram-se ativos no estudo das atividades antibacteriana com valores de CIM entre 6,25-100 µg/mL, leishmanicida (CI50 menores de 15 µg/mL) e esquistossomosse (CIM 12,5 µg/mL). Os testes de viabilidade celular mostraram uma baixa citotoxicidade do extrato e da fração com valores de concentrações superiores a 625 µg/mL e IC50 de 316 µg/mL. Os resultados obtidos forneceram dados relevantes em relação à composição química da espécie, avaliação biológica, assim como dos teores das saponinas presentes no extrato etanol 70 % das folhas de S. paniculatum.
Solanum paniculatum popularly known as jurubeba, is a species widely used in the Brazilian folk medicine for the treatment of liver and gastrointestinal disorders. Remarkably important it is the fact that is the only species of the genus Solanum that has been included in the Brazilian Health Unique System (SUS in Portuguese) list of medicinal plants of interest for the development of phytotherapic formulations. As part of our contribution toward understanding the uses and benefits derived from this species, the aim of the present work was the study of the chemical composition of the ethanolic 70 % extract of the leaves of S. paniculatum, through the identification, isolation and structural elucidation of the metabolites present in the extract. We also evaluated the cytotoxic activity of the alcoholic tincture (70 %) as well as, the antibacterial, antileishmanial, antischistossomal activities and cell viability test (normal cells) of the ethanolic 70 % extract and the enriched fraction of saponins (f’26). Different chromatographic techniques for isolation and purification were performed (Sephadex LH-20, High Performance Liquid Chromatography coupled to the Diode Array Detector (CLAE-DAD), High Performance Liquid Chromatography coupled to the Refractive Index Detector (HPLC-RI), which led to the isolation of 11 substances, that were characterized using spectroscopic techniques (one and two dimensional) and mass spectrometry. Fractionation of hydroalcoholic [ethanol (70%)] tincture led to the isolation of eight new steroidal saponins named: 6-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25S)-3β,6α,23-trihydroxy-5α-spirostane (1), 6-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25R)-3β,6α,23-trihydroxy-5α-spirostane (2), 6-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25S)-3β,6α,23-trihydroxy-5α-spirostane (3), 6-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25R)-3β,6α,23-trihydroxy-5α-spirostane (4), 3-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23S,25R)-3β,6α,23-trihydroxy-5α-spirostane (5), 3-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23S,25R)-3β,6α,23-trihydroxy-5α-spirostane (6), 6-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,25S)-1β,3β,6α-trihydroxy-5α-spirostane (7), 6-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,25S)-3β,4β,6α-trihydroxy-5α-spirostane (8), as well as rutin (9), caffeic acid (10) and caffeic anhydride (10a). Saponins (1-8) quantification in the 70% ethanol extract were performed by UPLC-ESI-IT-MS, this procedure allowed to determine the high content of saponins in the tincture (333.6 0,09 mg.g-1). The antitumor test for the ethanolic 70 % extract showed low activity against the cell tested with MIC values higher than 192.92 µg/mL, nonetheless, the ethanolic extract as well as the f’26 fraction were active in the study of antibacterial (MIC values ranging between 6.25-100 µg/mL), leishmanicidal (values of IC50 under 15 µg/mL) and schistosomicidal activities (MIC 12.5 µg/mL). The cell viability test showed a low cytotoxicity of ethanolic 70 % extract and the saponin fraction, with concentrations values up to 625 µg/mL and IC50 values from 316 µg/mL. The obtained results provided evidence regarding the chemical composition of the species, biological activities as well as the saponins content in the tincture.
Solanum paniculatum popularly known as jurubeba, is a species widely used in the Brazilian folk medicine for the treatment of liver and gastrointestinal disorders. Remarkably important it is the fact that is the only species of the genus Solanum that has been included in the Brazilian Health Unique System (SUS in Portuguese) list of medicinal plants of interest for the development of phytotherapic formulations. As part of our contribution toward understanding the uses and benefits derived from this species, the aim of the present work was the study of the chemical composition of the ethanolic 70 % extract of the leaves of S. paniculatum, through the identification, isolation and structural elucidation of the metabolites present in the extract. We also evaluated the cytotoxic activity of the alcoholic tincture (70 %) as well as, the antibacterial, antileishmanial, antischistossomal activities and cell viability test (normal cells) of the ethanolic 70 % extract and the enriched fraction of saponins (f’26). Different chromatographic techniques for isolation and purification were performed (Sephadex LH-20, High Performance Liquid Chromatography coupled to the Diode Array Detector (CLAE-DAD), High Performance Liquid Chromatography coupled to the Refractive Index Detector (HPLC-RI), which led to the isolation of 11 substances, that were characterized using spectroscopic techniques (one and two dimensional) and mass spectrometry. Fractionation of hydroalcoholic [ethanol (70%)] tincture led to the isolation of eight new steroidal saponins named: 6-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25S)-3β,6α,23-trihydroxy-5α-spirostane (1), 6-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25R)-3β,6α,23-trihydroxy-5α-spirostane (2), 6-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25S)-3β,6α,23-trihydroxy-5α-spirostane (3), 6-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23R,25R)-3β,6α,23-trihydroxy-5α-spirostane (4), 3-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23S,25R)-3β,6α,23-trihydroxy-5α-spirostane (5), 3-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,23S,25R)-3β,6α,23-trihydroxy-5α-spirostane (6), 6-O-α-L-rhamnopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,25S)-1β,3β,6α-trihydroxy-5α-spirostane (7), 6-O-β-D-xylopyranosyl-(1’’→3’)-β-D-quinovopyranosyl-(22S,25S)-3β,4β,6α-trihydroxy-5α-spirostane (8), as well as rutin (9), caffeic acid (10) and caffeic anhydride (10a). Saponins (1-8) quantification in the 70% ethanol extract were performed by UPLC-ESI-IT-MS, this procedure allowed to determine the high content of saponins in the tincture (333.6 0,09 mg.g-1). The antitumor test for the ethanolic 70 % extract showed low activity against the cell tested with MIC values higher than 192.92 µg/mL, nonetheless, the ethanolic extract as well as the f’26 fraction were active in the study of antibacterial (MIC values ranging between 6.25-100 µg/mL), leishmanicidal (values of IC50 under 15 µg/mL) and schistosomicidal activities (MIC 12.5 µg/mL). The cell viability test showed a low cytotoxicity of ethanolic 70 % extract and the saponin fraction, with concentrations values up to 625 µg/mL and IC50 values from 316 µg/mL. The obtained results provided evidence regarding the chemical composition of the species, biological activities as well as the saponins content in the tincture.
Descrição
Palavras-chave
Solanaceae, Saponina, Cromatografía líquida de alta eficiência, Leishmaniose, Citotoxicidade