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Horna Pinedo Vanessa.Pdf (1.825Mb) UNIVERSIDAD NACIONAL DE TRUJILLO FACULTAD DE FARMACIA Y BIOQUÍMICA ESCUELA PROFESIONAL DE FARMACIA Y BIOQUÍMICA TESIS II Actividad antiplasmodial in vitro de los alcaloides totales de los bulbos de Clinanthus incarnatus y Clinanthus ruber PARA OPTAR EL GRADO ACADÉMICO DE BACHILLER EN FARMACIA Y BIOQUÍMICA AUTORA: HORNA PINEDO, Madeleine Vanessa ASESORA: Dra. SOTO VÁSQUEZ, Marilú Roxana TRUJILLO – PERÚ 2020 Biblioteca Digital – Dirección de Sistemas de Informática y Comunicación DEDICATORIA A Dios por permitirme estar aquí, por la familia que me ha dado y por ser mi guía en el sendero de la vida. A mis padres A la unión cuyas enseñanzas han sido lumbre en el camino de mi formación. A mi madre, Aurora P. V, porque al igual que su nombre, al llegar cada día la encuentro presente para brindarme su apoyo y consejos, haciendo de mí una persona con valores y principios que orientan mi accionar; a ella por ser ejemplo de fortaleza, valentía, cariño y paciencia. A mi padre, Andrés H. V, porque a pesar de los infortunios, ha sabido cómo salir adelante y dar lo mejor a su familia, enseñándonos que con esfuerzo, constancia y disciplina A mis hermanos mayores podemos superar las dificultades, para alcanzar así nuestras metas. María, Omar, Paola y Liliana, por su apoyo en los buenos y malos momentos; a ustedes por cada hora, minuto, segundo compartido, llenos de risas, peleas, travesuras y más, ahora cada uno ha tomado su camino, pero aun así siempre estamos unidos por lazos inquebrantables. A mis sobrinos Biblioteca de FarmaciaRodrigo B. H. y M oyre lBioquímicaia V.H, por llegar a iluminar nuestras vidas y llenar de alegría a nuestra familia. “No está en las estrellas mantener nuestro destino, sino en nosotros mismos” – William Shakespeare Madeleine Vanessa Horna Pinedo Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajo la misma licencia 2.5 iPerú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital – Dirección de Sistemas de Informática y Comunicación AGRADECIMIENTO A Dios por ser base de nuestra moral, por permitirnos despertar cada día con salud, fuerzas y empeño para que cada momento sea de aprendizaje, de crecimiento personal y el tiempo necesario para que alcancemos las metas trazadas en nuestras vidas. A mi asesora Dra. Marilú Roxana Soto Vásquez, que con su amplia experiencia y conocimientos ha orientado al correcto desarrollo y culminación exitosa del presente trabajo de investigación. Quiero expresar mi más grande y sincero agradecimiento al Dr. Jaume Bastida, de la Universidad de Barcelona (España), por su valioso aporte durante todo éste proceso, permitiendo el desarrollo de este trabajo. Biblioteca de Farmacia y Bioquímica Al proyecto N° 110-2018-FONDECYT-CONCYTEC. “Proyectos de Investigación Básica y Aplicada 2018 – 01: «Bulbos Silvestres del Perú: Una Nueva Fuente de Productos Naturales Alcaloidales con Actividad Antiprotozoaria», por el apoyo económico brindado para el desarrollo del presente trabajo de investigación. Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajo la misma licencia 2.5i iPerú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital – Dirección de Sistemas de Informática y Comunicación PRESENTACIÓN SEÑORES MIEMBROS DEL JURADO: Dando cumplimiento a lo establecido por el reglamento de grados y títulos de la Facultad de Farmacia y Bioquímica de la Universidad Nacional de Trujillo, someto a vuestra honorable consideración y justo criterio el presente informe de Tesis II que lleva de título: Actividad antiplasmodial in vitro de los alcaloides totales de los bulbos de Clinanthus incarnatus y Clinanthus ruber. Es propicia ésta oportunidad para manifestar mi más sincero reconocimiento a mi alma máter y toda su plana docente, que con sus enseñanzas contribuyen en nuestra formación profesional de manera positiva y sólida. Señores miembros del jurado, dejo a vuestra consideración la respectiva calificación del presente trabajo y de antemano muy agradecida por el tiempo brindado. Trujillo, octubre del 2020 Biblioteca de Farmacia y Bioquímica HORNA PINEDO, Madeleine Vanessa DNI: 48509480 Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajo la misma licencia 2.5ii iPerú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital – Dirección de Sistemas de Informática y Comunicación JURADO DICTAMINADOR Dr. ZARI GIL, Gilmer (Presidente) Dr. RUIZ REYES, Segundo Guillermo (Miembro) Biblioteca de Farmacia y Bioquímica Dra. SOTO VÁSQUEZ, Marilú Roxana (Miembro) Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajo la misma licencia 2.5iv Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital – Dirección de Sistemas de Informática y Comunicación RESUMEN El objetivo del presente trabajo de investigación fue evaluar la actividad antiplasmodial in vitro de los alcaloides totales de los bulbos de Clinanthus incarnatus y Clinanthus ruber. Las plantas en estudio fueron recolectadas de los distritos de Pataz y Otuzco, respectivamente, en la región de La Libertad. A partir del extracto blando obtenido por maceración de los bulbos con metanol y mediante extracciones sucesivas basadas en cambios de pH y uso de solventes orgánicos se obtuvo el extracto de alcaloides totales. Luego se realizó la identificación cualitativa de los alcaloides mediante reacciones de coloración y precipitación, dando positivo a los reactivos de Dragendorf, Wagner y Mayer, mostrando precipitados de color anaranjado, marrón y blanco, respectivamente. Además, se realizó la cromatografía en capa fina, dando positivo al reactivo de Dragendorff. Para los extractos de Clinanthus incarnatus y Clinanthus ruber se obtuvo un porcentaje de rendimiento de alcaloides del 0,71% y 0,81%, respectivamente. La identificación y cuantificación de los alcaloides presentes en los bulbos se realizó mediante Cromatografía de Gases acoplada a Espectrometría de Masas (CG-EM), usando Galantamina como compuesto de referencia, la CG-EM mostró un total de 7 alcaloides identificados presentes en el bulbo de Clinanthus incarnatus siendo el más abundante licorina (19,73 ug gal/100 mg PS), seguido de galantina (10,36 ug gal/100 mg PS ), vitatina/crinina (10,22 ug gal/100 mg PS ), hipamina (10,16 ug gal/100 mg PS), 3-O-acetilpowellina (10,14 ug gal/100 mg PS), 11,12-deshidroanhidrolicorina (10,04 ug gal/100 mg PS) y 1-O-acetillicorina (9,85 ug gal/100 mg PS). En los bulbos de Clinanthus ruber se identificaron 8 alcaloides, siendo el más abundante licorina (70,2 ug gal/100 mg PS), seguido de anhidrolicorina (18 ug gal/100 mg PS), 11,12-deshidroanhidrolicorina (5,81 ug gal/100 mg PS), 2,4-didehidro-2- deshidroxilicorina (4,15 ug gal/100 mg PS), 8-O-dimetilmaritidina (3,45 ug gal/100 mg PS) e hipamina (Biblioteca0,1 ug gal/100 mg PdeS); a Farmaciasu vez se encontr ayro nBioquímica 2 alcaloides aun no identificados (m/z 125 tipo homolicorina y m/z 201, M=273). Los alcaloides totales de los bulbos de Clinanthus incarnatus y Clinanthus ruber presentaron actividad antiplasmoidal frente a Plasmodium falciparum (cepa FCR3 resistente a Cloroquina), con una inhibición de 90.1% y 94% y con CI50 de 0.375ug/mL y 0.241ug/mL respectivamente. Palabras Clave: Plasmodium falciparum, actividad antiplasmodial, Clinanthus incarnatus, Clinanthus ruber Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajo la misma licencia 2.5v Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital – Dirección de Sistemas de Informática y Comunicación ABSTRACT The objective of this research work was to evaluate the in vitro antiplasmodial activity of the total alkaloids of the bulbs of Clinanthus incarnatus and Clinanthus ruber. The plants under study were collected from the districts of Pataz and Otuzco, respectively, in the La Libertad region. The extract of total alkaloids was obtained from the soft extract obtained by maceration of the bulbs with methanol and by successive extractions based on changes in pH and use of organic solvents. Then the qualitative identification of the alkaloids was carried out by means of coloration and precipitation reactions, giving positive to the Dragendorf, Wagner and Mayer reagents, showing precipitates of orange, brown and white color, respectively. In addition, thin layer chromatography was performed, testing positive for Dragendorff's reagent. For the extracts of Clinanthus incarnatus and Clinanthus ruber, a percentage of alkaloid yield of 0.71% and 0.81%, respectively, was obtained. The identification and quantification of the alkaloids present in the bulbs was carried out by Gas Chromatography coupled to Mass Spectrometry (GC-MS), using Galantamine as a reference compound, the GC-MS showed a total of 7 identified alkaloids present in the bulb of Clinanthus incarnatus being the most abundant liquorine (19.73 ug gal / 100 mg PS), followed by galantin (10.36 ug gal / 100 mg PS), vitatin / crinine (10.22 ug gal / 100 mg PS), hypamine (10.16 ug gal / 100 mg PS), 3-O-acetylpowellin (10.14 ug gal / 100 mg PS), 11,12- dehydroanhydrolichorin (10.04 ug gal / 100 mg PS) and 1-O -acetylicorin (9.85 ug gal / 100 mg PS). In the Clinanthus ruber bulbs, 8 alkaloids were identified, the most abundant being liquorine (70.2 ug gal / 100 mg PS), followed by anhydrolichorin (18 ug gal / 100 mg PS), 11,12-dehydroanhydrolichorin (5.81 ug gal / 100 mg PS), 2,4-didehydro-2-dehydroxylicorin (4.15 ug gal / 100 mg PS), 8-O-dimethylmaritidine (3.45 ug gal / 100 mg PS) and hyipamine (0, 1 ug gal / 100 mg PS); in turn, 2 alkaloids were found not yet identified (m / z 125 homolicorinBiblioteca type and m / z 20 de1, M Farmacia= 273).
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