Identificación, establecimiento in vitro y análisis fitoquímico preliminar de especies silvestres de ñame (Dioscorea spp.) empleadas con fines medicinales

Título en ingles: Identification, in vitro establishment and preliminary phytochemical analysis of wild yam (Dioscorea spp.) used for medicinal purposesTítulo corto: Identificación, establecimiento in vitro y análisis fitoquímico preliminar de especies silvestres de ñameResumen:  Tubérculos del género Dioscorea comercializados con fines medicinales, fueron recolectados con el propósito de lograr su establecimiento a condiciones in vitro. Previamente se lograron identificar taxonómicamente las especies y por medio de análisis fitoquímicos demostrar su potencial farmacéutico. El material recolectado fue identificado como Dioscorea coriacea, D. lehmannii, D. meridensis, D. polygonoides y una especie comestible D. trifida. Tubérculos recolectados de centros de acopio y traídos de campo fueron lavados, desinfectados, asperjados con Ácido Giberélico (AG3) y sembrados en sustrato BM-2®, en invernadero a 18°C día y 10°C noche. Los tubérculos completos o por secciones fueron almacenados en bolsas herméticas a temperatura ambiente. Posteriormente se desinfectó material vegetal de las especies D. coriacea, D. lehmannii, D. meridensis y D polygonoides, seleccionando explantes de brotes sanos (D. coriacea / laboratorio) para su establecimiento. Se evaluaron tres medios de cultivo para establecimiento, el que presentó los mejores resultados fue Medio Murashige and Skoog (1962) suplementado con BAP 1 mL/L, AG3 1 mL/L y Putrescina 2 mL/L. Para la extracción y análisis de metabolitos secundarios se utilizaron tubérculos de D. coriacea, D. lehmannii y D. polygonoides, empleando como solvente de extracción metanol. Se  encontró mayor concentración de extracto vegetal en D. coriacea (54%), y mediante cromatografía en capa delgada (CCD), se confirmó la presencia de saponinas, que resultó mayor en comparación con D. polygonoides especie reconocida por su alto contenido de saponinas. Estos resultados permitirán realizar análisis más avanzados de los compuestos presentes y plantear su propagación masiva en condiciones in vitro. Palabras clave: diosgenina, micropropagación, ñame silvestre, cultivo de tejidos vegetales, saponinas, fitoquímica.Key words: diosgenin, micropropagation, wild yam, tissue culture, saponins, phitochemistry.Recibido: agosto 20 de 2014      Aprobado: abril 20 de 2015Wild tubers of the genus Dioscorea sold for medicinal use were collected for the purpose of achieving its establishment under in vitro conditions. First we taxonomically identified the species and through phytochemical analysis demonstrated pharmaceutical potential. The material collected was identified as Dioscorea coriacea, D. lehmannii, D. meridensis, D. polygonoides and the edible species D. trifida. Tubers collected from wholesale distributors and from the field were washed, disinfected, sprayed with Gibberellic Acid (GA3) and planted in substrate BM-2®, in a greenhouse at 18 ° C during the day and 10 ° C overnight. Whole tubers or sections thereof were stored in sealed bags at room temperature. Subsequently plant material of the species D. coriacea, D. lehmannii, D. meridensis and D. polygonoides was disinfected and healthy buds (D. coriacea / laboratory) were selected for in vitro establishment. Three different culture media were evaluated for establishment; that which presented the best results was the Murashige and Skoog (1962) medium, supplemented with BAP 1 mL / L, GA3 1 mL / L and Putrescin 2 mL / L. For the collection and analysis of secondary metabolites, tubers of D. coriacea, D. lehmannii and D. polygonoides were used, using methanol as the extraction solvent. The highest concentration of plant extract, 54%, was found in D. coriacea, a higher value than that of D. polygonoides, which had been reported previously; the presence of saponins was confirmed by thin layer chromatography (TLC). These results will enable more advanced analysis of the present compounds and enhance their mass propagation under in vitro conditions.Key words: diosgenin, micropropagation, wild yam, tissue culture, saponins, phitochemistry

Estudio fitoquímico y evaluación de la actividad antimalárica de Piper cumanense y Piper holtonii

El uso terapéutico de plantas medicinales es una práctica común en regiones del mundo donde se presenta la malaria y desempeña un papel importante en la prevención y tratamiento de la enfermedad en zonas endémicas. En Colombia se estima que anualmente se presentan más de 60.000 casos de malaria (INS) y en algunas regiones del país se hace uso de preparaciones con plantas medicinales que incluyen especies del género Piper para tratar la enfermedad. En el presente trabajo se realizó la evaluación de la actividad antimalárica in vitro e in vivo de los extractos etanólicos y de fracciones de diferente polaridad de las especies vegetales Piper cumanense y Piper holtonii (Piperaceae), lo que permitió identificar fracciones con actividad antiplasmódica y aislar metabolitos secundarios del grupo de los fenilpropanoides, flavonoides, ácidos benzoicos, esteroides y terpenos. Adicionalmente se caracterizaron microscópicamente las hojas de las dos especies, realizando cortes transversales y longitudinales. De P. cumanense se aisló el cromeno ácido gaudichaudiánico, el ácido 4-metoxi-3-(3metil-2-butenil)-benzoico, la 2’,4’,6’-trihidroxidihidrochalcona y 3 flavonoides C-glicósidos. Además se aislaron β-sitosterol, estigmasterol y β-sitosterol-3-O-glucósido y se detectaron por CG-MS 6 terpenoides. De P. holtonii se aislaron los fenilpropanoides apiol y dilapiol como metabolitos mayoritarios y los flavonoides kaempferol-3-O-rutinósido y rutina, así como β-sitosterol y sitosterol-3-O-β-glucósido.Abstract. The therapeutic use of medicinal plants is a common practice in endemic from malaria regions in the world and plays an important role in the prevention and treatment of disease. In Colombia is estimated to occur annually more than 60,000 cases of malaria and in some regions it herbal preparations are used to treat the disease. In this work, in vitro and in vivo evaluation of antimalarial activity of ethanolic extracts and different fractions of P. cumanense and P. holtonii (Piperaceae) results in the identification of active antiplasmodial fractions and also in the isolation of secondary metabolites as phenylpropanoids, flavonoids, benzoic acids, steroids and terpenes. Additionally the leaves of the two species were characterized microscopically. In P. cumanense was isolated gaudichaudianic acid, 4-methoxy-3-(3-methyl-2-butenyl)benzoic, 2 ', 4', 6'-trihidroxydihydrochalcone and C-glycosides flavonoids. In addition were isolated β-sitosterol, stigmasterol and β-sitosterol-3-O-glucoside and were detected by GC-MS 6 terpenoids. In P. holtonii was isolated as major metabolites, phenylpropanoids, apiol and dillapiole, and flavonoids kaempferol-3-O-rutinoside, rutine, sitosterol and β-sitosterol-3-O-βglucoside.Doctorad

Identificación, establecimiento in vitro y análisis fitoquímico preliminar de especies silvestres de ñame (Dioscorea spp.) empleadas con fines medicinales

Wild tubers of the genus Dioscorea sold for medicinal use were collected for the purpose of achieving its establishment under in vitro conditions. First we taxonomically identified the species and through phytochemical analysis demonstrated pharmaceutical potential. The material collected was identified as Dioscorea coriacea, D. lehmannii, D. meridensis, D. polygonoides and the edible species D. trifida. Tubers collected from wholesale distributors and from the field were washed, disinfected, sprayed with Gibberellic Acid (GA3) and planted in substrate BM-2®, in a greenhouse at 18 ° C during the day and 10 ° C overnight. Whole tubers or sections thereof were stored in sealed bags at room temperature. Subsequently plant material of the species D. coriacea, D. lehmannii, D. meridensis and D. polygonoides was disinfected and healthy buds (D. coriacea / laboratory) were selected for in vitro establishment. Three different culture media were evaluated for establishment; that which presented the best results was the Murashige & Skoog (1962) medium, supplemented with BAP 1 mL / L, GA3 1 mL / L and Putrescin 2 mL / L. For the collection and analysis of secondary metabolites, tubers of D. coriacea, D. lehmannii and D. polygonoides were used, using methanol as the extraction solvent. The highest concentration of plant extract, 54%, was found in D. coriacea, a higher value than that of D. polygonoides, which had been reported previously; the presence of saponins was confirmed by thin layer chromatography (TLC). These results will enable more advanced analysis of the present compounds and enhance their mass propagation under in vitro conditions.Tubérculos del género Dioscorea comercializados con fines medicinales, fueron recolectados con el propósito de lograr su establecimiento a condiciones in vitro. Previamente se lograron identificar taxonómicamente las especies y por medio de análisis fitoquímicos demostrar su potencial farmacéutico. El material recolectado fue identificado como Dioscorea coriacea, D. lehmannii, D. meridensis, D. polygonoides y una especie comestible D. trifida. Tubérculos recolectados de centros de acopio y traídos de campo fueron lavados, desinfectados, asperjados con Ácido Giberélico (AG3) y sembrados en sustrato BM-2®, en invernadero a 18°C día y 10°C noche. Los tubérculos completos o por secciones fueron almacenados en bolsas herméticas a temperatura ambiente. Posteriormente se desinfectó material vegetal de las especies D. coriacea, D. lehmannii, D. meridensis y D polygonoides, seleccionando explantes de brotes sanos (D. coriacea / laboratorio) para su establecimiento. Se evaluaron tres medios de cultivo para establecimiento, el que presentó los mejores resultados fue Medio Murashige & Skoog (1962) suplementado con BAP 1 mL/L, AG3 1 mL/L y Putrescina 2 mL/L. Para la extracción y análisis de metabolitos secundarios se utilizaron tubérculos de D. coriacea, D. lehmannii y D. polygonoides, empleando como solvente de extracción metanol. Se encontró mayor concentración de extracto vegetal en D. coriacea (54%), y mediante cromatografía en capa delgada (CCD), se confirmó la presencia de saponinas, que resultó mayor en comparación con D. polygonoides especie reconocida por su alto contenido de saponinas. Estos resultados permitirán realizar análisis más avanzados de los compuestos presentes y plantear su propagación masiva en condiciones in vitro

Identification, in vitro establishment and preliminary phytochemical analysis of wild yam (Dioscorea spp.) used for medicinal purposes

Wild tubers of the genus Dioscorea sold for medicinal use were collected for the purpose of achieving its establishment under in vitro conditions. First we taxonomically identified the species and through phytochemical analysis demonstrated pharmaceutical potential. The material collected was identified as Dioscorea coriacea, D. lehmannii, D. meridensis, D. polygonoides and the edible species D. trifida. Tubers collected from wholesale distributors and from the field were washed, disinfected, sprayed with Gibberellic Acid (GA3) and planted in substrate BM-2®, in a greenhouse at 18 ° C during the day and 10 ° C overnight. Whole tubers or sections thereof were stored in sealed bags at room temperature. Subsequently plant material of the species D. coriacea, D. lehmannii, D. meridensis and D. polygonoides was disinfected and healthy buds (D. coriacea / laboratory) were selected for in vitro establishment. Three different culture media were evaluated for establishment; that which presented the best results was the Murashige & Skoog (1962) medium, supplemented with BAP 1 mL / L, GA3 1 mL / L and Putrescin 2 mL / L. For the collection and analysis of secondary metabolites, tubers of D. coriacea, D. lehmannii and D. polygonoides were used, using methanol as the extraction solvent. The highest concentration of plant extract, 54%, was found in D. coriacea, a higher value than that of D. polygonoides, which had been reported previously; the presence of saponins was confirmed by thin layer chromatography (TLC). These results will enable more advanced analysis of the present compounds and enhance their mass propagation under in vitro conditions