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Pontificia Universidad Javeriana Facultad De Ciencias Programa De Postgrado

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Pontificia Universidad Javeriana Facultad De Ciencias Programa De Postgrado PONTIFICIA UNIVERSIDAD JAVERIANA FACULTAD DE CIENCIAS PROGRAMA DE POSTGRADO ESTUDIO FITOQUÍMICO Y SCREENING DE LA ACTIVIDAD BIOLÓGICA DE LAS ESPECIES Conyza trihecatactis y Ageratina vacciniaefolia PAOLA BORREGO MUÑOZ TRABAJO DE GRADO Presentado como requisito parcial para optar el título de MAGISTER EN CIENCIAS BIOLÓGICAS PONTIFICIA UNIVERSIDAD JAVERIANA FACULTAD DE CIENCIAS PROGRAMA DE POSGRADOS BOGOTÁ D.C 2017 NOTA DE ADVERTENCIA "La Universidad no se hace responsable por los conceptos emitidos por sus alumnos en sus trabajos de tesis. Solo velará por que no se publique nada contrario al dogma y a la moral católica y porque las tesis no contengan ataques personales contra persona alguna, antes bien se vea en ellas el anhelo de buscar la verdad y la justicia". Artículo 23 de la Resolución Nº13 de julio de 1946. ESTUDIO FITOQUÍMICO Y SCREENING DE LA ACTIVIDAD BIOLÓGICA DE LAS ESPECIES Conyza trihecatactis y Ageratina vacciniaefolia PAOLA BORREGO MUÑOZ APROBADO ____________________ _______________________ ______________________ Dr. JUAN MARTÍNEZ PhD. Dr. ANTONIO GUZMÁN MSc. Dra. GINA MÉNDEZ PhD. ESTUDIO FITOQUÍMICO Y SCREENING DE LA ACTIVIDAD BIOLÓGICA DE LAS ESPECIES Conyza trihecatactis y Ageratina vacciniaefolia PAOLA BORREGO MUÑOZ ____________________________ ________________________________ Dra. Concepción Judith Puerta Dra. Alba Alicia Trespalacios Decana Directora de Posgrados Facultad de Ciencias Facultad de Ciencias AGRADECIMIENTOS A Dios por darme la fuerza, por permitirme una vez más hacer mis sueños realidad y haber puesto en mi camino aquellas personas que han sido mi soporte durante todo este tiempo. A la Pontificia Universidad Javeriana, a mi Director Jorge Robles por sus valiosos aportes durante la realización de este trabajo, a mis compañeros de laboratorio por su amistad y cooperación constante. A la Fundación Universitaria Juan N. Corpas, a la Rectoria, Vicerrectoría Académica y Administrativa por contribuir a la realización de los ensayos antiinflamatorios. A mi Co-director Miguel Pombo por la confianza que siempre ha depositado en mí y por demostrarme que con amor al trabajo y constancia se alcanzan grandes logros. A la Clínica Juan N. Corpas, al Departamento de Patología por la realización del estudio histopatológico. A la Universidad Militar Nueva Granada, al Doctor Ericsson Coy quien me acogió y me brindo su apoyo cuando lo necesité, a los integrantes del Grupo de Investigación InQuiBio porque siempre tuvieron una sonrisa para conmigo. A la Universidad Distrital Francisco José de Caldas, al Profesor Javier Matulevich por la toma de muestras en el equipo de EM, por su apoyo y consejos. A toda mi familia por el amor y el apoyo incondicional, a todos aquellos que han contribuido, directa o indirectamente a la realización de este trabajo. GRACIAS A TODOS POR TODO!!! TABLA DE CONTENIDO Página RESUMEN .............................................................................................................. 1 INTRODUCCIÓN .................................................................................................... 4 OBJETIVOS ............................................................................................................ 5 OBJETIVO GENERAL ............................................................................................ 5 OBJETIVOS ESPECÍFICOS ................................................................................... 5 1. ESTADO DEL ARTE ....................................................................................... 6 1.1 FAMILIA ASTERACEAE .......................................................................... 6 1.1.1 Características generales de la familia Asteraceae ................................ 6 1.1.2 Distribución de la familia Asteraceae ....................................................... 7 1.1.3 Quimiotaxonomía de la familia Asteraceae ............................................. 7 1.2 Género Conyza ........................................................................................ 8 1.2.1 Distribución del género Conyza ............................................................... 8 1.2.2 Usos etnobotánicos del género Conyza .................................................. 9 1.2.3 Actividad biológica de especies del género Conyza ............................... 9 1.2.4 Estudios fitoquímicos del género Conyza .............................................. 10 1.3 Especie Conyza trihecatactis ............................................................... 13 1.3.1 Características morfológicas de la especie Conyza trihecatactis ......... 13 1.3.2 Distribución de la especie Conyza trihecatactis .................................... 15 1.3.3 Usos etnobotánicos de la especie Conyza trihecatactis ....................... 15 1.3.4 Actividad biológica de la especie Conyza trihecatactis ......................... 15 1.3.5 Estudios fitoquímicos de la Conyza trihecatactis .................................. 16 1.4 Género Ageratina .................................................................................. 17 1.4.1 Distribución del género Ageratina .......................................................... 17 1.4.2 Usos etnobotánicos del género Ageratina ............................................. 18 1.4.3 Actividad biológica del género Ageratina ............................................... 18 1.4.4 Estudios fitoquímicos del género Ageratina .......................................... 19 1.5 Especie Ageratina vacciniaefolia......................................................... 22 1.5.1 Características morfológicas de la especie Ageratina vacciniaefolia ... 22 1.5.2 Distribución de la especie Ageratina vacciniaefolia .............................. 22 1.5.3 Actividad biológica de la especie Ageratina vacciniaefolia ................... 23 1.5.4 Estudios fitoquímicos de la especie Ageratina vacciniaefolia ............... 24 1.6 Técnicas de uso frecuente para la determinación de metabolitos secundarios ...................................................................................................... 24 1.6.1 Cromatografía en Capa Fina (CCF) ....................................................... 25 1.6.2 Cromatografía líquida acoplada a Espectrometría de Masas (HPLC- MS) ................................................................................................................. 25 1.6.3 Espectrometría de masas (MS) ............................................................. 25 1.6.4 Espectroscopia de Resonancia Magnética Nuclear (RMN) .................. 26 1.7 Actividad antiinflamatoria .................................................................... 27 1.7.1 Inflamación aguda .................................................................................. 28 1.7.2 Inflamación crónica ................................................................................. 29 1.7.3 Modelos de evaluación in vivo de la actividad antiinflamatoria ............ 30 1.8 Generalidades del Cáncer .................................................................... 31 1.8.1 Características y biología de las células tumorales .............................. 33 1.8.2 Ciclo celular tumoral ............................................................................... 33 1.8.3 Epidemiología del Cáncer ...................................................................... 34 1.8.4 Ensayos in vitro para evaluar agentes antitumorales............................ 35 1.8.5 Muerte celular ......................................................................................... 36 1.9 Especies reactivas del oxígeno ........................................................... 38 1.9.1 Mecanismo de defensa antioxidante ..................................................... 39 1.9.2 Determinación de la capacidad antioxidante ......................................... 40 1.9.3 Ensayo del DPPH• (1,1-difenil-2-picril-hidrazilo) ................................... 41 1.10 Toxicidad aguda en Artemia salina ..................................................... 42 2. METODOLOGÍA ............................................................................................ 43 2.1 Generalidades ....................................................................................... 43 2.2 Métodos de separación cromatográfica .............................................. 43 2.3 Técnicas para la elucidación estructural y caracterización de los compuestos aislados ...................................................................................... 44 2.3.1 Espectrometría de masas (MS) ............................................................. 44 2.3.2 Espectroscopia de Resonancia Magnética Nuclear (RMN) .................. 44 2.3.3 Cromatografía líquida acoplada a Espectrometría de Masas (HPLC- DAD-MS) ............................................................................................................ 44 2.3.4 Cromatografía líquida de longitud de onda variable (semipreparativa) (HPLC-VWD (semiprep)) ................................................................................... 45 2.4 Estudio fitoquímico de las partes aéreas de las especies vegetales Conyza trihecatactis y Ageratina vacciniaefolia ........................................... 46 2.4.1 Recolección e identificación del material vegetal .................................. 46 2.4.2 Obtención de extractos ........................................................................... 46 2.4.3 Obtención de fracciones para Conyza trihecatactis .............................. 47 2.4.4 Obtención de
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