Potencial antifúngico y tamizaje fitoquímico del extracto acetónico de las hojas de Momordica charantia L. sobre Candida albicans
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Date
2022
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Universidad Nacional de Trujillo
Abstract
El objetivo de este trabajo de investigación fue determinar el potencial antifúngico del extracto de las hojas de Momordica charantia L. sobre cepas de Cándida albicans. La planta se recolectó de las zonas rurales del centro poblado de Cerro Blanco, distrito de Laredo, Provincia de Trujillo, Región la Libertad. Luego de la recolección las hojas que fueron sometidos a secado en la estufa a 45ºC hasta llegar a peso constante, seguidamente se preparó el extracto por el método de maceración durante 7 días con acetona. Posteriormente los extractos se solubilizaron en solución de dimetilsulfóxido al 1 % a una concentración de 2048 μg/ mL y apartir de ella se aplicaron diluciones seriadas para la obtención de concentraciones hasta 16 μg /mL en cada pocillo de las microplacas; previamente se hizo la reactivación de la cepa de Candida albicans ATCC mediante el cultivo en caldo Sabouraud y Agar Sabouraud lo cual fue llevado a estufa a 37°C por 24 horas. Para la estandarización de los inóculos fúngicos se llevó hasta una turbidez equivalente a la escala de McFarland 0.5 (aproximadamente 108 UFC/ mL) hasta un inoculo fúngico final de 104 UFC / ml que se utilizó en todos los ensayos .El ensayo para la determinación de la concentración inhibitoria mínima (CIM) se realizó mediante la metodología recomendada por el Clinical and Laboratorial Standard Institute donde se colocó en 96 pocillos de microplacas 100 µL de medio de cultivo de caldo Sabouraud para levadura, 20 µL de la sustancia de prueba (extractos), en concentraciones seriadas de 2048 a 16 μg / ml, y 80 μL de suspensiones fúngicas (104 UFC / mL); finalmente se obtuvo que el extracto acetónico de hojas de charantia L. tiene un potencial antifúngico bajo contra Cándida albicans. En los últimos ensayos que se realizó se empleó el método de la “prueba de la gota “donde se preparó 3 extractos que fueron, fracción diclorometano, etanólica y acuoso-ácida; donde se identificó metabolitos por el método de coloración.
ABSTRACT The objective of this research work was to determine the antifungal potential of Momordica charantia L. leaf extract on Candida albicans strains. The plant was collected from the rural areas of the town of Cerro Blanco, district of Laredo, Province of Trujillo, La Libertad Region. After harvesting the leaves, they were subjected to drying in the oven at 45ºC until reaching constant weight, then the extract was prepared by the maceration method for 7 days with acetone. Subsequently, the extracts were solubilized in a 1% dimethyl sulfoxide solution at a concentration of 2048 μg/mL, and from there, serial dilutions were applied to obtain concentrations up to 16 μg/mL in each well of the microplates; Previously, reactivation of the Candida albicans ATCC strain was carried out by growing it in Sabouraud broth and Sabouraud Agar, which was taken to an oven at 37°C for 24 hours. For the standardization of the fungal inocula, it was brought to a turbidity equivalent to the McFarland scale 0.5 (approximately 108 CFU/mL) to a final fungal inoculum of 104 CFU/mL that was used in all the tests. The test for the determination of the minimum inhibitory concentration (MIC) was performed using the methodology recommended by the Clinical and Laboratorial Standard Institute where 100 µL of Sabouraud broth culture medium for yeast, 20 µL of the test substance (extracts) were placed in 96 wells of microplates , in serial concentrations from 2048 to 16 μg/ml, and 80 μL of fungal suspensions (104 CFU/ml); Finally, it was obtained that the acetone extract of Momordica charantia L. leaves has a low antifungal potential against Candida albicans. In the last tests that were carried out, the "drop test" method was used, where 3 extracts were prepared: dichloromethane, ethanolic and aqueous-acid fractions; where metabolites were identified by the staining method.
ABSTRACT The objective of this research work was to determine the antifungal potential of Momordica charantia L. leaf extract on Candida albicans strains. The plant was collected from the rural areas of the town of Cerro Blanco, district of Laredo, Province of Trujillo, La Libertad Region. After harvesting the leaves, they were subjected to drying in the oven at 45ºC until reaching constant weight, then the extract was prepared by the maceration method for 7 days with acetone. Subsequently, the extracts were solubilized in a 1% dimethyl sulfoxide solution at a concentration of 2048 μg/mL, and from there, serial dilutions were applied to obtain concentrations up to 16 μg/mL in each well of the microplates; Previously, reactivation of the Candida albicans ATCC strain was carried out by growing it in Sabouraud broth and Sabouraud Agar, which was taken to an oven at 37°C for 24 hours. For the standardization of the fungal inocula, it was brought to a turbidity equivalent to the McFarland scale 0.5 (approximately 108 CFU/mL) to a final fungal inoculum of 104 CFU/mL that was used in all the tests. The test for the determination of the minimum inhibitory concentration (MIC) was performed using the methodology recommended by the Clinical and Laboratorial Standard Institute where 100 µL of Sabouraud broth culture medium for yeast, 20 µL of the test substance (extracts) were placed in 96 wells of microplates , in serial concentrations from 2048 to 16 μg/ml, and 80 μL of fungal suspensions (104 CFU/ml); Finally, it was obtained that the acetone extract of Momordica charantia L. leaves has a low antifungal potential against Candida albicans. In the last tests that were carried out, the "drop test" method was used, where 3 extracts were prepared: dichloromethane, ethanolic and aqueous-acid fractions; where metabolites were identified by the staining method.
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Keywords
Momordica charantia L., Cándida albicans