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Universidad Técnica Particular Deloja Área Biológica Y Biomédica UNIVERSIDAD TÉCNICA PARTICULAR DELOJA La Universidad Católica de Loja ÁREA BIOLÓGICA Y BIOMÉDICA TITULO DE BIOQUÍMICO FARMACÉUTICO Aislamiento, Identificación y Evaluación de la actividad antimicrobiana de metabolitos secundarios aislados de la planta medicinal Siparuna eggersii y de sus extractos. TRABAJO DE TITULACIÓN AUTORA: Vélez Ramírez, Vilma Elizabeth DIRECTOR: Ramírez Robles, Jorge Yandry, PhD LOJA-ECUADOR 2016 APROBACIÓN DEL DIRECTOR DE TRABAJO DE TITULACIÓN Ph.D Jorge Yandry Ramírez Robles DOCENTE DE LA TITULACIÓN De mi consideración: El presente trabajo de titulación Aislamiento, Identificación y Evaluación de la actividad antimicrobiana de metabolitos secundarios aislados de la planta medicinal Siparuna eggersii y de sus extractos realizado por Vélez Ramírez Vilma Elizabeth ha sido orientado y revisado durante su ejecución, por cuanto se aprueba la presentación del mismo. Loja, 22 de Septiembre del 2016 f) Jorge Yandry Ramírez Robles ii DECLARACIÓN DE AUTORÍA Y CESIÓN DE DERECHOS “Yo Vélez Ramírez Vilma Elizabeth declaro ser autora del presente trabajo de titulación Aislamiento, Identificación y Evaluación de la actividad antimicrobiana de metabolitos secundarios aislados de la planta medicinal Siparuna eggersii y de sus extractos, de la titulación de Bioquímica y Farmacia, siendo Jorge Yandry Ramírez Robles director del presente trabajo; y eximo expresamente a la Universidad Técnica Particular de Loja y a sus representantes legales de posibles reclamos o acciones legales. Además certifico que las ideas, conceptos, procedimientos y resultados vertidos en el presente trabajo investigativo, son de mi exclusiva responsabilidad. Adicionalmente declaro conocer y aceptar la disposición del Art. 88 del Estatuto Orgánico de la Universidad Técnica Particular de Loja que en su parte pertinente textualmente dice: “Forman parte del matrimonio de la Universidad la propiedad intelectual de investigaciones, trabajos científicos o técnicos y tesis de grado o trabajos de titulación que se realicen con el apoyo financiero, académico o institucional (operativo) de la Universidad” f. Autor: Vélez Ramírez Vilma Elizabeth Cédula: 1106039215 iii DEDICATORIA A Dios porque he visto su infinito amor y gracia cada día de mi vida, por sus abundantes bendiciones y por darme un nuevo propósito para mi vida. A mis padres Franklin y Esthela por su amor y apoyo incondicional, por guiarme con sabiduría y regalarme su confianza en cada etapa de mi vida. A mis hermanos Diego y Jairo por compartir su vida y amistad conmigo y por su apoyo e interés durante mi carrera. Vilma Elizabeth Vélez Ramírez iv AGRADECIMIENTOS Agradezco a Dios por darme vida y sabiduría para lograr con éxito mis metas, y por mover para mi cada obstáculo que se presentó durante mi carrera universitaria. A mis queridos padres por todo su amor, respeto, consejos y esfuerzo por darme siempre lo mejor y ser mi ejemplo a seguir. A mis amigas Maddie y Rachel así como a mi grupo alfa y omega por su interés y apoyo durante el desarrollo de este proyecto incluso más allá de las fronteras. Al PhD Jorge Ramírez por compartir conmigo sus conocimientos, por su importante aporte, profesionalismo, y sabiduría al guiar este proyecto de investigación. A mis amigos y compañeros que durante estos cinco años me han brindado su amistad y de una u otra forma han aportado en mi formación profesional y personal. Vilma Elizabeth Vélez Ramírez v ÍNDICE DE CONTENIDOS CARÁTULA ............................................................................................................. i APROBACIÓN DEL DIRECTOR DE TRABAJO DE TITULACIÓN......................... ii DECLARACIÓN DE AUTORÍA Y CESIÓN DE DERECHOS .................................. iii DEDICATORIA ...................................................................................................... iv AGRADECIMIENTOS ............................................................................................. v LISTA DE FIGURAS ............................................................................................. viii LISTA DE TABLAS ................................................................................................ ix ABREVIATURAS .................................................................................................... x RESUMEN .............................................................................................................. 1 ABSTRACT ............................................................................................................ 2 INTRODUCCIÓN .................................................................................................... 3 CAPITULO I ............................................................................................................... 4 MARCO TEÓRICO .................................................................................................... 4 1.1 Antecedentes ................................................................................................... 5 1.2 Plantas Medicinales .......................................................................................... 5 1.3 Plantas Medicinales del Ecuador .................................................................. 5 1.3.1 Familia Siparunaceae. .......................................................................................................... 6 1.3.2 Genero Siparuna. .................................................................................................................... 6 1.3.3 Siparuna eggersii. ................................................................................................................... 7 1.3.4 Taxonomía de Siparuna eggersii. ................................................................................... 8 1.4 Metabolitos Secundarios .................................................................................. 8 1.4.1 Terpenos. .................................................................................................................................... 8 1.4.2 Flavonoides................................................................................................................................ 9 1.5 Fitoquímica ..................................................................................................... 10 1.6 Técnicas Cromatográficas .............................................................................. 10 1.6.1 Cromatografía de Columna (CC). ................................................................................. 11 1.6.2 Cromatografía de capa fina (CCF). .............................................................................. 11 1.6.3 Cromatografía de Gases acoplada a Espectrometría de Masas . ................ 11 1.7 Resonancia magnética nuclear (RMN) ....................................................... 12 1.8 Actividad Antimicrobiana ................................................................................ 12 1.8.1 Bacterias. .................................................................................................................................. 12 CAPITULO II ............................................................................................................ 14 MATERIALES Y METODOS ................................................................................... 14 2.2 Recolección del Material Vegetal .................................................................... 16 vi 2.3 Obtención de los Extractos ............................................................................. 16 2.4 Análisis de los Extractos ................................................................................. 16 2.5 Fraccionamiento en Cromatografía en Columna ............................................ 16 2.6 Cromatografía de Capa Fina .......................................................................... 16 2.7 Unión de Fracciones ....................................................................................... 17 2.8 Purificación de Compuestos .......................................................................... 17 2.8.1 Fracción VV36/7. ................................................................................................................... 17 2.8.2 Fracción VV38/7. ................................................................................................................... 17 2.8.3 Fracción VV39/7. ................................................................................................................... 18 2.9 Factor de Retención (Rf) ................................................................................ 18 2.10 Identificación de los Compuestos ................................................................. 18 2.10.1 Cromatografía de Gases Acoplada a Espectrometría de Masas. ............... 18 2.10.2 Resonancia Magnética Nuclear (RMN) ................................................................... 19 2.11 Determinación de la Actividad Antimicrobiana .............................................. 19 2.11.1 Microorganismos de Prueba. ........................................................................................ 19 2.11.2 Preparación de la Muestra. ............................................................................................ 19 2.11.3 Preparación del Cultivo Bacteriano. .......................................................................... 19 2.11.4 Preparación de la suspensión de inóculos para bacterias. ............................ 20 2.11.5 Ensayo de Concentración Mínima Inhibitoria (CMI).
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