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1 EVALUACIÓN DEL POTENCIAL ANTIOXIDANTE DE MEZCLAS DE EXTRACTOS ETANÓLICOS DE Mollinedia Racemosa, Siparuna Sessiliflora Y

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1 EVALUACIÓN DEL POTENCIAL ANTIOXIDANTE DE MEZCLAS DE EXTRACTOS ETANÓLICOS DE Mollinedia Racemosa, Siparuna Sessiliflora Y EVALUACIÓN DEL POTENCIAL ANTIOXIDANTE DE MEZCLAS DE EXTRACTOS ETANÓLICOS DE Mollinedia racemosa, Siparuna sessiliflora y Croton leptostachyus Trabajo de grado presentado como requisito parcial para optar al título de Biólogo CLAUDIA CRISTINA PÉREZ JARAMILLO Director WALTER MURILLO ARANGO Doctor en Ciencias Químicas Codirector WILMER FERNANDO SÁNCHEZ PERALTA Licenciado en Educación Básica con Énfasis en Ciencias Naturales y Educación Ambiental UNIVERSIDAD DEL TOLIMA FACULTAD DE CIENCIAS PROGRAMA DE BIOLOGÍA IBAGUÉ – TOLIMA 2014 1 2 DEDICATORIA Este trabajo está dedicado a mi familia, especialmente a mi madre, Amparo Jaramillo Cardona, ella es el pilar de mi vida y quien ha luchado incansablemente para brindarme la mejor educación, por ella he logrado este objetivo que es solo el inicio de grandes propósitos. 3 AGRADECIMIENTOS Son muchas las personas que me han acompañado en esta aventura que inicié hace unos años. Gracias a ellas, en parte, soy lo que soy. Ahora ha llegado el momento de cerrar este capítulo, pero no puedo hacerlo sin antes agradecerles todo lo que me han brindado… A mis directores, Dr. Jonh Jairo Méndez Arteaga y Dr. Walter Murillo Arango por permitir mi incorporación en su grupo de investigación con total libertad y autonomía y por confiar en mí y apoyarme siempre. Especial agradecimiento al Grupo de Investigación en Productos Naturales de la Universidad del Tolima (GIPRONUT), a los compañeros y profesores del área de Química; a los profesores Giann Carlos Peñaloza, Cesar Jaramillo, Antoni Rueda, Elizabeth Murillo; al Licenciado Carlos Martín Guerra; a los Biólogos Ángel Jiménez, Laura Daniela Rodríguez, Diego Oliveros, y a mis compañeros tesistas y pasantes Iván Cupitra, Lina Dávila, Francisco Santamaría y Lorena Osorio. A los integrantes del Laboratorio de Servicios y Extensión de análisis químico (LASEREX); a la Oficina de Investigaciones y Desarrollo Científico de la Universidad del Tolima, al departamento de Química, al programa de Biología de la Facultad de Ciencias Básicas y a todas las personas que en el transcurso del proyecto hicieron parte activa en la ejecución de este. No hay que olvidar a la profesora Julia Ester Gallego, docente de la Facultad de Ciencias de la salud; quien se interesó en mi trabajo y contribuyo con sus conocimientos a enriquecerlo. Agradecer especialmente a Lic. Wilmer Fernando Sánchez Peralta, codirector de este trabajo y compañero de laboratorio incondicional, por apoyarme en todo momento y darme ánimos para continuar. A mi familia: padres, abuelos, tíos, primos… que sin saber muy bien lo que he estado haciendo me han ayudado en los momentos difíciles. Agradecer especialmente a mis padres y hermanos. A los primeros, por haberme proporcionado siempre los mejores apoyos emocionales y materiales, por haberme educado en esos valores en los que me siento tan orgullosa y estar siempre a mi lado. A mis hermanos por su cariño y apoyo. GRACIAS A TODOS 4 CONTENIDO INTRODUCCIÓN ..................................................................................................................... 175 1. JUSTIFICACIÓN ............................................................................................................ 17 2. PLANTEAMIENTO DEL PROBLEMA ....................................................................... 19 3. OBJETIVOS .................................................................................................................... 21 3.1. OBJETIVO GENERAL ....................................................................................................... 21 3.2. OBJETIVOS ESPECÍFICOS ............................................................................................. 21 4. MARCO TEORICO ........................................................................................................ 22 4.1. GENERALIDADES DE LA FAMILIA MONIMIACEAE .................................................. 22 4.1.1. Descripción de las plantas del genero Mollinedia. .................................................... 26 4.2. GENERALIDADES DE LA FAMILIA SIPARUNACEAE ............................................... 28 4.2.1. Descripción de las plantas del género Siparuna ........................................................ 31 4.3. GENERALIDADES DE LA FAMILIA EUPHORBIACEAE ............................................ 34 4.3.1. Descripción de las plantas del genero Croton ........................................................... 37 4.4. RADICALES LIBRES ..................................................................................................... 39 4.4.1. Fuentes endógenas de Radicales Libres .................................................................... 41 4.4.2. Fuentes exógenas de Radicales Libres ...................................................................... 42 4.5. ANTIOXIDANTES .......................................................................................................... 42 4.5.1. Sistemas de defensa antioxidantes. ............................................................................ 43 5 4.5.1.1. Sistemas de defensa antioxidantes endógenos ................................................. 43 4.5.1.2. Sistema de defensa antioxidante exógeno ......................................................... 44 4.6. ACTIVIDAD ANTIOXIDANTE EN UN MODELO DE CELULAS SANGUINEAS (ERITROCITOS)......................................................................................................................... 47 4.7. ANÁLISIS DE INTERACCIÓN MEDIANTE ISOBOLOGRAMAS .......................... 48 5. DISEÑO METODOLÓGICO ......................................................................................... 51 5.1. MATERIALES ..................................................................................................................... 51 5.1.1. Obtención del material vegetal ..................................................................................... 51 5.1.2. Reactivos .......................................................................................................................... 53 5.2. MÉTODOS........................................................................................................................... 53 5.2.2. Caracterización preliminar de los extractos vegetales .............................................. 54 5.3. PRUEBA DE ACTIVIDAD ANTIOXIDANTE IN VITRO ................................................ 54 5.3.1. Actividad inhibitoria del catión radical del ácido 2,2- azinobis-(3 etilbenzotiazolina- 6-sulfónico ABTS) ...................................................................................................................... 54 5.3.2. Actividad estabilizante del radical 2,2-diphenil-1-picrilhidrazil (DPPH) ................. 55 5.4. ACTIVIDAD ANTIOXIDANTE DE MEZCLAS DE MOSQUERO (Croton leptostachyus), ROMADIZO (Mollinedia racemosa) Y LIMÓN DE MONTE (Siparuna sessiliflora). ................................................................................................................................. 56 5.5. ACTIVIDAD ANTIOXIDANTE EN UN MODELO DE CÉLULAS SANGUÍNEAS (ERITROCITOS)......................................................................................................................... 56 5.5.1. Obtención de los eritrocitos ........................................................................................... 56 5.5.2. Inhibición de la Hemolisis (Actividad antioxidante) .................................................... 56 6. RESULTADOS Y DISCUSIÓN .................................................................................... 58 6.1. CARACTERIZACIÓN DE LOS EXTRACTOS VEGETALES .................................. 58 6 6.2. ACTIVIDAD ANTIOXIDANTE IN VITRO DE LOS EXTRACTOS CRUDOS ........ 60 6.2.1. Actividad estabilizante del radical 2,2-diphenil-1-picrilhidrazil (DPPH). ................. 60 6.2.2. Actividad estabilizante del catión radical del ácido 2,2-azinobis-(3 etilbenzotiazolina-6-sulfónico (ABTS.+) ................................................................................... 62 6.3. ACTIVIDAD ESTABILIZANTE DPPH DE MEZCLAS DE EXTRACTOS DE M. racemosa, S. sessiliflora y C.leptostachyus .......................................................................... 63 6.4. ACTIVIDAD INHIBITORIA DEL RADICAL ABTS.+ DE MEZCLAS DE EXTRACTOS PROVENIENTES DE M. racemosa, S. sessiliflora y C.leptostachyus .... 66 6.5. ACTIVIDAD ANTIOXIDANTE EN UN MODELO DE CELULAS SANGUÍNEAS (ERITROCITOS)......................................................................................................................... 69 7. CONCLUSIONES .......................................................................................................... 73 RECOMENDACIONES ............................................................................................................. 74 REFERENCIAS .......................................................................................................................... 75 7 LISTA DE FIGURAS Figura 1. Distribución geografica de la familia Monimiaceae ......................... 23 Figura 2. Características morfológicas de la familia Monimiaceae ................ 24 Figura 3. Especie del genero Mollinedia (Mollinedia schottiana) ................... 27 Figura 4. Distribución geográfica de la familia Siparunaceae ........................ 29 Figura 5. Características morfológicas de la un espécimen de la familia Siparunaceae .......................................................................................... 30 Figura 6. Especie del género
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