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UPS-TTQ075.Pdf UNIVERSIDAD POLITÉCNICA SALESIANA SEDE QUITO CARRERA: INGENIERÍA EN BIOTECNOLOGÍA DE LOS RECURSOS NATURALES Trabajo de titulación previo a la obtención del título de: INGENIERA EN BIOTECNOLOGÍA DE LOS RECURSOS NATURALES. TEMA: “EVALUACIÓN DE LA ACTIVIDAD ANTIOXIDANTE DE LOS EXTRACTOS ETANÓLICOS DE CINCO ESPECIES DE ORQUÍDEAS DE LOS GÉNEROS Maxillaria, Catasetum y Epidendrum, PRESENTES EN EL ECUADOR” AUTORA: LISBETH JUDITH MOLINA SANDOVAL DIRECTOR: MARCO FERNANDO CERNA CEVALLOS Quito, agosto del 2020 Dedicatoria Primero a Dios por permitirme estar aquí y ahora, por brindarme salud, sabiduría, entendimiento y hacer que en muchas ocasiones la vida me sonría por sus bendiciones. A mi familia, por estar siempre presente, especialmente a mi papito Luis Molina por ser el pilar de mi vida, mi ejemplo y fortaleza a mi hermana Tamara, por apoyarme y aconsejarme a mi madre que aunque esté ausente, sé que nos tiene presentes en su corazón, ustedes me dan la fuerza necesaria para continuar y las ganas de luchar y salir a delante. A mi papito Pio Baschirotto por todo su apoyo incondicional, por escucharme, también por aconsejarme y sobre todo por ser el abuelo paterno que no tuve. A mi licenciada Ana Rosa Vásconez, por todo su apoyo, consejos y ánimo para no rendirme durante todo mi proceso universitario y fuera de él. A todos mis amigos, amigas y familiares quienes me han apoyado de una u otra forma. A todos aquellos apasionados investigadores y conservacionistas que buscan el rescate del conocimiento ancestral, integrando la investigación en ese proceso que da forma a la farmacia viva, dónde las plantas en su estado natural constituyen nuestra medicina del día a día. Lisbeth. Agradecimiento A Dios por permitirme concluir esta etapa de mi vida. A la Universidad Politécnica Salesiana por el financiamiento del Trabajo de Titulación a través del proyecto de investigación: ―Estudio taxonómico, propiedades antibacteriales y anticancerígenas de especies seleccionadas de los géneros Caucaea, Gongora, Bulbophylum, Dichaea, Epidendrum, Huntleya, Maxillaria, Sobralia, Stelis (Orchidaceae)‖ que se desarrolló en la UPS durante el período 2019, con el grupo Nunkui Wakan. A mi tutor de tesis PhD. Marco Cerna, por permitirme conocer el maravilloso mundo de las orquídeas, también por su guía, apoyo y asesoría. Al PhD. Paco Noriega, por compartirme sus conocimientos, por también guiarme y aconsejarme en el desarrollo de esta investigación. A Priscilla Muriel, investigadora de la PUCE, Gabriel Sáenz, biólogo de la PUCE, por su apoyo, consejos, compartirme información valiosa y conectar en la idea de la farmacia viva despertando mi ánimo e interés en este campo de investigación. Al investigador Luis Vaquero, por también compartirme sus conocimientos e información valiosa para desarrollar esta investigación. A la Ing. Erika Paredes, Ing. Ely Yugsi, Ing. Byron Fuertes, por compartirme sus conocimientos y guía, pero sobre todo por brindarme su amistad y consejo. A mis amigos y amigas: Lari Jaramillo, Bel Montaluisa, Adri Mogrovejo, Yaha Proaño, Martí Rojas, Andy Villarreal, Esteban Rojas, Francisco Hoyos, Danny Montoya, Viky Sánhez, Chris Quinchuela, Isra Vaca, Lili Tambaco, Tere Carbonell, Fulvio Cabrera, con quienes he compartido, conocimiento, proyectos, consejos, pero sobretodo esos momentos geniales de amistad que te animan a continuar y van abriendo nuevos caminos y oportunidades en la vida. Índice Introducción .........................................................................................................................1 Capítulo 1 ..............................................................................................................................3 Marco conceptual .................................................................................................................3 1.1. Familia Orchidaceae .................................................................................................3 1.1.1. Generalidades ......................................................................................................3 1.1.2. Importancia y usos de las orquídeas .....................................................................3 1.1.3. Descripción de la familia Orchidaceae .................................................................4 1.1.4. Descripción taxonómica ......................................................................................8 1.1.5. Género Maxillaria Ruiz & Pav. ...........................................................................8 1.1.6. Género Catasetum Rich. ex Kunth..................................................................... 10 1.1.7. Género Epidendrum L. ...................................................................................... 11 1.1.8. Presencia de orquídeas en el Ecuador ................................................................ 12 1.1.9. Etnobotánica de las orquídeas ............................................................................ 12 1.1.9.1. Orquídeas utilizadas en la medicina tradicional ecuatoriana ....................... 12 1.1.10. Estudios moleculares de las orquídeas ............................................................... 15 1.2. Fitoquímica de las orquídeas .................................................................................. 15 1.2.1. Metabolitos secundarios .................................................................................... 15 1.2.1.1. Fenoles ............................................................................................................. 16 1.2.1.3. Flavonoides ...................................................................................................... 17 1.2.1.4. Alcaloides ........................................................................................................ 17 1.2.2. Métodos de extracción de metabolitos ............................................................... 18 1.2.2.1. Método de Maceración .............................................................................. 18 1.2.3. Tipos de extractos botánicos .............................................................................. 19 1.2.3.1. Extractos fluidos ........................................................................................ 19 1.2.4. Métodos de análisis químico de metabolitos ............................................................ 19 1.2.4.1. Tamizaje fitoquímico ....................................................................................... 19 1.2.4.2. Espectrofotometría UV-visible ......................................................................... 19 1.3. Antioxidantes en orquídeas .................................................................................... 19 1.3.2. Métodos de determinación de antioxidantes ............................................................ 20 1.3.2.1. Metodología DPPH .......................................................................................... 20 Capítulo 2 ............................................................................................................................ 21 Marco metodológico ........................................................................................................... 21 2.1. Valoración etnobotánica .............................................................................................. 21 2.1.1. Colección de material vegetal .................................................................................. 21 2.1.2. Identificación botánica ............................................................................................ 23 2.1.2.1. Identificación morfológica................................................................................ 23 2.1.2.2. Identificación molecular ................................................................................... 23 2.2. Estudio fitoquímico ...................................................................................................... 26 2.2.1. Tratamiento del material vegetal ............................................................................. 26 2.2.2. Obtención de los extractos ...................................................................................... 26 2.2.2. Tamizaje fitoquímico .............................................................................................. 27 2.2.2.1. Determinación de fenoles y taninos .................................................................. 27 2.2.2.2. Determinación de flavonoides .......................................................................... 28 2.2.2.3. Determinación de alcaloides ............................................................................. 28 2.2.3. Cuantificación espectrofotométrica de metabolitos .................................................. 29 2.2.3.1. Cuantificación de fenoles totales ...................................................................... 29 2.2.3.2. Cuantificación de flavonoides totales................................................................ 32 2.3. Evaluación de la actividad antioxidante ..................................................................... 35 2.3.1. Preparación de la solución de DPPH ....................................................................... 35 2.3.2. Preparación de los estándares de Ácido ascórbico (vitamina C) ............................... 35 2.3.3. Procedimiento para la evaluación de antioxidantes en estándares y extractos ........... 36 2.3.3.1. Lectura de las soluciones .................................................................................
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