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Guzmanmoralesanapaola1d2.Pdf MAESTRIA En CIENCIAS^ Aumentarías UNIVERSIDAD VERACRUZANA INSTITUTO DE CIENCIAS BÁSICAS Actividad antioxidante y antimicrobiana del aceite esencial y extractos (crudo, acuoso, y etanólico) de Pipicha (Porophyllum tagetoides) TESIS Que para obtener el grado de: MAESTRO EN CIENCIAS ALIMENTARIAS Presenta: Q.F.B. Ana Paola Guzmán Morales Director de tesis: INSTITUTO QE MIAS B'SUTAS. Dra. Maribel Jiménez Fernández IJNIVÜRSIÜAÜ VI.-ACHUCHA BIBLIOTÍAT Xalapa, Veracruz Septiembre, 2009 DEDICATORIAS A Dios, por haberme bendecido con la oportunidad de realizar este posgrado, por haberme dado fuerzas para concluirlo, por la gente que utilizaste para enviarme un mensaje de apoyo y de aliento para seguir adelante y por sostenerme y cuidarme en todo momento, mostrando que no hay adversidad si vas a mi lado. A mis padres, Sr. José de Jesús Guzmán Salazar y Sra. Lucia Inés Morales Y Salazar, por su amor, dirección y cuidados. Gracias por su comprensión y paciencia, sin ustedes no hubiera podido terminar este ciclo. A mis hermanos, Ana Patricia Guzmán Morales e Isalas de Jesús Guzmán Morales, por su apoyo y cariño incondicional. A mis tías, Inés Andrade López, Yolanda Jiménez González y Patricia Guzmán Salazar, gracias por su cariño y sus consejos. A mis amigas, Yesenia, Diana, Itzel, Lucina, Judy, Lolita y Elva, por su amistad incondicional, apoyo y sinceridad. Gracias por sus oraciones, y por su paciencia para escucharme y estar a mi lado. A Mateo y Martha Estrada, gracias por sus oraciones y sus palabras de aliento. Finalmente, a todos aquellos que me apoyaron en oración para que este trabajo de investigación llegara a su fin y resultara lo mejor posible. AGRADECIMIENTOS Este trabajo fue realizado gracias al apoyo otorgado por las siguientes instituciones: • Instituto de Ciencias Básicas de la Universidad Veracruzana. • Hospital Escuela de Ginecobstetricia de la Universidad Veracruzana. • Facultad de Ingeniería Agroindustrial y de Alimentos de la Universidad de las Américas, Puebla. • Facultad de Química Farmacéutica Biológica (Q.F.B.) de la Universidad Veracruzana. e Unidad de Servicios de Apoyo en Resolución Analítica (SARA) de la Universidad Veracruzana. Un agradecimiento muy especial a la Dra. Maribel Jiménez Fernández, por su apoyo, tiempo y dedicación para que este proyecto se llevará a cabo. Gracias por su confianza y por creer en mí. A todos los profesores que contribuyeron a mi crecimiento profesional durante este proceso de aprendizaje, les agradezco todas sus enseñanzas, en especial al Dr. Ebner Azuara Nieto, al Dr. Erick Romeo Silva Hernández, al Dr. Iñigo Verdalet Guzmán, al Dr. Oscar García Barradas y la M.C. Yolanda Cocotle Ronzón. Agradezco a mi distinguido jurado por sus valiosas aportaciones que enriquecieron este trabajo de investigación. e Dr. Micloth López del Castillo-Lozano. ® Dra. Elia Nora Aquino Bolaños. • Dra. Guadalupe Luna Solano. ® Dr. Erick Romeo Silva Hernández. ÍNDICE GENERAL RESUMEN SUMMARY I. INTRODUCCIÓN.................................................................................... 1 II. MARCO TEÓRICO................................................................................. 4 2.1 Herbolaria en México....................................................................... 4 2.2 Pipicha (Porophyllum tagetoides).............. .................... .............. 4 2.2.1 Origen y clasificación................... ......................................... 4 2.2.2 Morfología y distribución..................... .... ...................... 5 2.2.3 Importancia económica.......................................................... 7 2.2.3 Usos........................................................................................ 8 2.3 Deterioro de los alimentos.............................. ............................... 8 2.3.1 Factores que intervienen en la alteración del alimento....... 9 2.3.2 Conservación de los alimentos................... ....... ........... 10 2.3.3 Aditivos alimenticios........................................................... 11 2.4. Radicales libres................................................................................ 12 2.4.1 Clasificación............................................................................. 13 2.4.2 Origen...................................................................................... 14 2.4.3 Funciones................................................................................ 15 2.4.4 Efectos adversos en la célula.............................................. 15 2.4.5 Mecanismos de acción de los radicales libres....................... 17 2.4.6 Acción de los radicales en los alimentos................................. 17 2.5. Antioxidantes.................................................................................... 18 2.5.1 Clasificación............................................................................ 18 2.5.2 Origen....................................................................................... 20 2.5.3 Actividades pro-oxidantes....................................................... 21 2.5.4 Efecto de los antioxidantes en los alimentos........................ 21 2.5.5 Métodos de laboratorio para el análisis de antioxidantes en los alimentos............................................................................. 21 2.6. Deterioro de los alimentos provocado por microorganismos........ 24 2.6.1 Métodos antimicrobianos de conservación de alimentos..... 33 2.6.2 Antimicrobianos........................................................................ 34 2.6.2.1 Antimicrobianos naturales.............................................. 35 2.6.2.2 Antimicrobianos naturales de origen vegetal................. 35 2.6.2.2.1 Aceites esenciales......................................... 35 2.6.3 Acción de los antimicrobianos en los microorganismos....... 37 2.6.4 Antioxidantes como antimicrobianos...................................... 37 2.7 Antecedentes........................................................... 38 2.8 Situación actual................... 39 III. PLANTEAMIENTO DEL PROBLEMA................................................. 40 IV. OBJETIVOS E HIPÓTESIS............................ ....................... 41 4.1 Objetivo general................................................................................ 41 4.2 Objetivos específicos....................................... 41 4.3 Hipótesis.................................. 42 V. MATERIAL Y MÉTODOS............................................................. 43 5.1 Material biológico...... ............................... 43 5.1.1 Pipicha (Porophyllum tagetoides) ........ 43 5.1.2 Cepas microbianas estudiadas.............................................. 44 5.1.2.1 Bacterias.............................................................................. 44 5.1.2.2 Levaduras.............................................................................. 44 5.1.2.3 Mohos.................................................................................... 44 5.2 Metodología....................................................................................... 45 5.2.1 Caracterización de pipicha, sus extractos y su emulsión... 46 5.2.1.1 Determinación de humedad de la planta completa y de las hojas........................................................ 46 5.2.1.2 Extracción y rendimiento de la emulsión de los extractos..................................................................... 47 5.2.1.2.1 Elaboración de los extractos para pruebas preeliminares............................................... 47 5.2.1.2.2 Elaboración de los extractos acuoso y etanólico.................................................... 47 5.2.12.3 Elaboración del extracto crudo................ 47 5.2.1.2.4 Extracción de la emulsión......................... 48 5.2.12.5 Determinación de rendimiento.................. 48 5.2.13 Barrido de la planta..................................................... 49 5.2.14 Análisis de compuestos volátiles............................... 49 5.2.15 Propiedades fisicoquímicas........................................ 50 5.2.15.1 Peso especifico........................................... 50 5.2.15.2 índice de refracción y grados Brix............. 50 5.2.15.3 Color............................................................ 51 5.2.2 Determinación de la actividad antioxidante.............. ........... 53 5.2.2.1 Cuantificación de compuestos con actividad antioxidante............................................................ 53 5.2.2.11 Densidad óptica........................ 53 5.2.2.12 Determinación de fenoles totales...,........... 53 5.2.2.13 Cuantificación de carotenos.......... ............. 53 5.2.2.14 Cuantificación de vitamina C ,,........... 64 5.2.2.2 Análisis de óxido-reducción............................ ......... 56 5.2.2.2.1 Poder reductor............................................. 56 5.22.2.2 Potencial Redox.......................................... 56 5.2.2.3 Porcentaje de captura del radical DPPH................... 57 5.2.3 Determinación de la actividad antimicrobiana........................ 58 5.2.3.1 Preparación, cultivo y ajuste de la concentración de los microorganismos de prueba............................... 58 5.2.3.11 Preparación de los microorganismos de prueba.......................................................... 58 5.2.3.12 Preparación del inoculo............................... 58 5.2.3.13 Escala de Me. Farland................................ 59 5.2.3.2 Pruebas de actividad antimicrobiana para bacterias y levaduras................................................................
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