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Universidad Técnica Particular De Loja Àrea UNIVERSIDAD TÉCNICA PARTICULAR DE LOJA La Universidad católica de Loja ÀREA BIOLÓGICA TITULO DE BIOQUÍMICA Y FARMACIA Obtención de extractos, aislamiento y caracterización de metabolitos secundarios de la especie Erythrina smithiana. TRABAJO DE TITULACIÓN AUTOR: Aguilar Jaramillo, Hermes Leonardo DIRECTOR: Romero Benavides, Juan Carlos, Ing. LOJA – ECUADOR 2015 Esta versión digital, ha sido acreditada bajo la licencia Creative Commons 4.0, CC BY-NY- SA: Reconocimiento-No comercial-Compartir igual; la cual permite copiar, distribuir y comunicar públicamente la obra, mientras se reconozca la autoría original, no se utilice con fines comerciales y se permiten obras derivadas, siempre que mantenga la misma licencia al ser divulgada. http://creativecommons.org/licenses/by-nc-sa/4.0/deed.es Septiembre, 2015 APROBACIÓN DEL DIRECTOR DEL TRABAJO DE TITULACIÓN Ingeniero. Juan Carlos Romero Benavides. DOCENTE DE LA TITULACIÓN De mi consideración: El presente trabajo de titulación: “Obtención de extractos, aislamiento y caracterización de metabolitos secundarios de la especie Erythrina smithiana con actividad antiparasitaria”, realizado por Hermes Leonardo Aguilar Jaramillo, ha sido orientado y revisado durante su ejecución, por cuanto se aprueba la presentación del mismo. Loja, septiembre de 2015 f)____________________________ Ing. Romero Benavides Juan Carlos C.I. 1103018477 ii DECLARACIÓN DE AUTORÍA Y CESIÓN DE DERECHOS. Yo, Hermes Leonardo Aguilar Jaramillo, declaro ser autor del presente trabajo de titulación: “Obtención de extractos, aislamiento y caracterización de metabolitos secundarios de la especie Erythrina smithiana con actividad antiparasitaria”, de la titulación de Bioquímica y Farmacia, siendo el Ing. Juan Carlos Romero Benavides, 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 patrimonio de la Universidad la propiedad intelectual de investigaciones, trabajos científicos o técnicos, o tesis de grado que se realicen a través, o con el apoyo financiero, académico e institucional (operativo) de la Universidad. f)____________________________ Hermes Leonardo Aguilar Jaramillo C.I. 1104966641 iii DEDICATORIA El presente trabajo de investigación va dedicado a DIOS por haberme dado salud, para poder lograr este objetivo propuesto, además por brindarme su infinidad bondad y amor. A mis padres GEOVANNY Y GREYZ por haberme apoyado en todo momento por todos sus consejos, valores, por toda esa motivación, por sus ejemplos de perseverancia y constancia y sobre todo por todo el amor brindado. A mi hija LUCIANA MARTINA por ser el principal motivo de terminar mi carrera profesional. A GABRIELA por todo su apoyo y compañía durante toda la carrera. A mis hermanos TATIANA SALOME y RONNY SEBASTIAN por brindarme todo su cariño y apoyo día a día. A mi padre de crianza JACINTO JUVENAL que fue un pilar fundamental en mi formación tanto espiritual como humana y por ver en él, un ser humano luchador y emprendedor. Hermes Leonardo Aguilar Jaramillo. iv AGRADECIMIENTO Mi agradecimiento va muy especialmente a mis padres, por su apoyo durante estos años, por sus consejos para poder salir adelante y ser un hombre de bien, además por su paciencia para poder culminar mi carrera profesional. Agradezco al Ing. Juan Carlos Romero B. por brindarme su amistad, confianza y conocimientos y por toda la orientación brindada para el normal desarrollo de este trabajo de investigación. Al Departamento de Química Aplicada de la Universidad Técnica Particular de Loja, y a cada uno de los que lo integran, por su confianza, amistad y apoyo brindado durante el tiempo que se empleó durante esta investigación. A todos mis compañeros y amigos que de una u otra manera, fueron personas importantes, estos años de vida estudiantil, agradezco a todos ellos por compartir momentos incomparables; por haberme ayudado y por haberme motivado día a día. A la Dra. Alirica Suarez, por impartirme sus conocimientos referentes a la identificación de compuestos. A los Bioquímicos Ronald Silva y Paola Castillo por su ayuda y sus conocimientos en las técnicas de aislamiento de metabolitos secundarios. Gracias. v Tabla de contenido APROBACIÓN DEL DIRECTOR DEL TRABAJO DE TITULACIÓN .............................. ii DECLARACIÓN DE AUTORÍA Y CESIÓN DE DERECHOS. ........................................ iii DEDICATORIA ............................................................................................................ iv AGRADECIMIENTO ..................................................................................................... v LISTA DE TABLAS ..................................................................................................... viii LISTA DE FIGURAS .................................................................................................... ix ABREVIATURAS .......................................................................................................... x RESUMEN .................................................................................................................... 1 ABSTRACT .................................................................................................................. 2 INTRODUCCION .......................................................................................................... 3 CAPÍTULO I. ................................................................................................................. 5 1. Fin, propósito y componentes del proyecto. ........................................................... 6 1.1. Fin del proyecto. ............................................................................................. 6 1.2. Propósito del proyecto. ................................................................................... 6 1.3. Componentes del proyecto. ............................................................................ 6 CAPÍTULO II. ................................................................................................................ 7 2. Aspectos generales. .............................................................................................. 8 2.1 Antecedentes y biodiversidad. ........................................................................ 8 2.2 Fitoquímica. .................................................................................................. 10 2.3 Metabolitos secundarios. .............................................................................. 10 2.4 Descripción del genero Erythrina. ................................................................. 11 2.5 Descripción de la especie Erythrina smithiana Krukoff. ................................. 13 2.6 Cromatografía. .............................................................................................. 14 2.6.1 Cromatografía en capa fina (CCF). ........................................................ 15 2.6.2 Cromatografía columna. ........................................................................ 15 2.6.3 Cromatografía de gases acoplada a espectroscopia de masas (CG- EM)………………………… .................................................................................. 15 2.7 Resonancia magnética nuclear (RMN). ........................................................ 16 2.8 Cristalización. ............................................................................................... 16 2.9 Punto de fusión. ............................................................................................ 17 CAPITULO III. ............................................................................................................. 18 3. METODOLOGÍA .................................................................................................. 19 3.1 Selección y recolección de la materia vegetal. .............................................. 19 3.2 Obtención de extractos. ................................................................................ 20 3.3 Desclorofilación del extracto. ........................................................................ 21 vi 3.4 Fraccionamiento del extracto desclorofilado. ................................................ 21 3.4.1 Cromatografía en columna. ................................................................... 21 3.4.2 Cromatografía en capa fina. .................................................................. 22 3.5 Microcolumna. .............................................................................................. 22 3.6 Unión y purificación. ..................................................................................... 22 3.7 Caracterización de metabolitos secundarios. ................................................ 22 3.7.1 Punto de fusión. ..................................................................................... 22 3.7.2 Solubilidad. ............................................................................................ 23 3.7.3 Factor de retención. ............................................................................... 23 3.7.4 Cromatografía de gases acoplado a espectroscopia de masas (CG/EM). 24 3.7.5 Espectroscopia
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