Pulicaria Arabica (L.) Cass. Et Rhanterium Adpressum Coss

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Pulicaria Arabica (L.) Cass. Et Rhanterium Adpressum Coss REPUBLIQUE ALGERIENNE DEMOCRATIQUE ET POPULAIRE MINISTERE DE L’ENSEIGNEMENT SUPERIEUR ET DE LA RECHERCHE SCIENTIFIQUE UNIVERSITE LARBI BEN M’HIDI -OUM El BOUAGHI- FACULTE DES SCIENCES EXACTES ET DES SCIENCES DE LA NATURE ET DE LA VIE DEPARTEMENT DES SCIENCES DE LA NATURE ET DE LA VIE N° d’ordre :………… N° de série :………… MEMOIRE présenté pour l’obtention du diplôme de Magister En BIOCHIMIE APPLIQUEE Option: Biochimie des produits naturels d’origine végétale Thème : Extraction des métabolites secondaires de plantes médicinales: Pulicaria arabica (L.) Cass. et Rhanterium adpressum Coss. & Durieu. Et evaluation de leurs propriétés bioactives Présenté par : Djermane Nadia Soutenu le 30 juin 2014 devant le jury : Président: Pr. Snoussi Med Mourad UNIVERSITE LARBI BEN M’HIDI-OEB Rapporteur: Pr. Gherraf Noureddine UNIVERSITE LARBI BEN M’HIDI-OEB Examinateur: Pr. Zellagui Amar UNIVERSITE LARBI BEN M’HIDI-OEB Examinateur: Dr. Arhab Rabeh UNIVERSITE LARBI BEN M’HIDI-OEB Année universitaire 2013-2014 SOMMAIRE REMERCIEMENT-DEDICACE……………………………………………………………………………………………………. I LISTE DES ABRIVIATIONS……………………………………………………………………………………………………….. II LISTE DES TABLEAUX……………………………………………………………………………………………………………… III LISTE DES FIGURES………………………………………………………………………………………………………………… VI INTRODUCTION…………………………………………………………………………………………………………………….. 1 ETUDE BIBLIOGRAPHIQUE I. Les métabolites secondaires……………………………………………………………………………………………….. I.1.Introduction………………………………………………………………………………………………………………………….. 3 I.2.Classification…………………………………………………………………………………………………………………………. 3 I.2.1.Les Composés phénoliques……………………………………………………………………………………………….. 4 I.2.1.1.Définition……………………………………………………………………………………………………………………….. 4 I.2.1.2.Classification…………………………………………………………………………………………………………………… 4 I.2.1.2.1.Les flavonoïdes……………………………………………………………………………………………………………. 4 I.2.1.2.2.Les tanins…………………………………………………………………………………………………………………….. 5 I.2.1.2.3.Les acides phénoliques………………………………………………………………………………………………… 7 I.2.1.2.4.Les stilbènes………………………………………………………………………………………………………………… 9 I.2.1.2.5.Les lignanes…………………………………………………………………………………………………………………. 12 I.2.2.Les alcaloïdes…………………………………………………………………………………………………………………..... 12 I.2.2.1.Définition……………………………………………………………………………………………………………………….. I.2.2.2.Classification…………………………………………………………………………………………………………………… 13 II. Les huiles essentielles………………………………………………………………………………………………………… 13 II.1.Définition…………………………………………………………………………………………………………………………….. 13 II.2.Répartition systématique et localisation…………………………………………………………………………….. 15 II.3.Toxicité des composants des huiles essentielle……………………………………………………………………. 15 II.4.Caractéristiques physico-chimiques…………………………………………………………………………………….. 15 II.4.1.propriétés physiques………………………………………………………………………………………………………… 16 II.4.2.Composition chimique………………………………………………………………………………………………………. 17 II.5.Méthodes d’extraction………………………………………………………………………………………………………… 17 II.5.1.Distillation………………………………………………………………………………………………………………………… 17 II.5.2.L’expression……………………………………………………………………………………………………………………… 23 II.5.3.Extraction par des solvants……………………………………………………………………………………………….. II.5.4.Extraction par fluide supercritique……………………………………………………………………………………. 23 II.5.5.Extraction par micro-ondes………………………………………………………………………………………………. 24 II.6.Méthodes d’identification chimiques…………………………………………………………………………………… 25 II.6.1.La chromatographie en phase gazeuse……………………………………………………………………………… 26 II.6.2.La spectrométrie de masse……………………………………………………………………………………………….. 27 II.6.3.Couplage CPG/MS…………………………………………………………………………………………………………….. 27 II.7.Propriétés……………………………………………………………………………………………………………………………. 27 II.7.1.Activité antimicrobienne…………………………………………………………………………………………………… 28 II.7.1.1.Méthodes d’évaluation de l’activité antimicrobienne……………………………………………………. 29 II.7.1.1.1.Aromatogramme………………………………………………………………………………………………………… 29 II.7.1.1.2.Méthode de diffusion en puits……………………………………………………………………………………. 30 II.7.1.1.3.Méthode de dilution………………………………………………………………………………………. 32 II.7.1.1.4.Méthode en microatmosphère……………………………………………………………………… 32 II.7.2.Activité antioxydante…………………………………………………………………………………………… 32 II.7.2.1.Définition des radicaux libres……………………………………………………………………………. 33 II.7.2.2.Définition des antioxydants……………………………………………………………………………….. 33 II.7.2.3.Types d’antioxydants…………………………………………………………………………………………. 34 II.7.2.4.Méthodes d’évaluation de l’activité antioxydante…………………………………………….. 34 II.7.2.4.1.Test de DPPH………………………………………………………………………………………………….. 34 II.7.2.4.2.Test de décoloration du β-carotène……………………………………………………………….. 35 II.7.2.4.3.Test FRAP (Ferric Reducing Antioxidant Power)……………………………………………… 35 III. Les plantes médicinales sélectionnées…………………………………………………………………. 37 III.1.La famille Astéracée……………………………………………………………………………………………….. 37 III.1.1.Etymologie………………………………………………………………………………………………………….. 37 III.1.2.Classification systématique…………………………………………………………………………………. 37 III.1.3.Répartition géographique……………………………………………………………………………………. 37 III.1.4.Caractéristiques générales…………………………………………………………………………………… 37 III.1.5.Place des Astéracées dans la médecine traditionnelle…………………………………………. 38 III.2.La plante Rhanterium adpressum………………………………………………………………………….. 39 III.2.1.Description………………………………………………………………………………………………………….. 39 III.2.2.Position systématique…………………………………………………………………………………………. 40 III.2.3.Répartition géographique……………………………………………………………………………………. 40 III.2.4.Utilisation en médecine traditionnelle………………………………………………………………… 41 III.2.5.Travaux phytochimiques antérieurs…………………………………………………………………….. 41 III.2.5.1.Sur le genre………………………………………………………………………………………………………. 41 III.2.5.2.Sur l’espèce………………………………………………………………………………………………………. 42 III.1.3.La plante Pulicaria arabica…………………………………………………………………………………. 45 III.1.3.1.Description……………………………………………………………………………………………………….. 45 III.1.3.2.Position systématique………………………………………………………………………………………. 46 III.1.3.3.Répartition géographique………………………………………………………………………………… 46 III.1.3.4.Utilisation en médecine traditionnelle……………………………………………………………… 47 III.1.3.5.Travaux phytochimiques antérieurs…………………………………………………………………. 47 III.1.3.5.1.Sur le genre…………………………………………………………………………………………………… 47 III.1.3.5.1.Sur l’espèce…………………………………………………………………………………………………… 48 ETUDE EXPERIMENTALE IV. Matériels et méthodes…………………………………………………………………………………………. 51 IV.1.Matériels……………………………………………………………………………………………………………. 51 IV. 1.1.Choix du matériel végétal…………………………………………………………………………………… 51 IV.1.2.Appareillage et produits chimiques……………………………………………………………………… 51 IV.1.3.Origine et choix des souches microbiennes…………………………………………………………. 51 IV.1.4.Choix des milieux de culture……………………………………………………………………………….. 51 IV.2.Méthodes…………………………………………………………………………………………………………… 52 IV.2.1.Extraction de l’huile essentielle des plantes………………………………………………………… 53 IV.2.1.1.Procédé d’extraction………………………………………………………………………………………… 53 IV.2.1.2.Conservation des huiles essentielles obtenues…………………………………………………. 53 IV.2.1.3.Détermination du rendement d’extraction………………………………………………………. 53 IV.2.1.4. Analyse de la composition chimique des huiles essentielles par CG/SM………….. 54 IV.2.2. Préparation des extraits bruts des plantes…………………………………………………………. 54 IV.2.3.Evaluation de l'activité antimicrobienne……………………………………………………………… 55 IV.2.3.1.Préparation des dilutions…………………………………………………………………………………. 55 IV.2.3.2.Préparation des disques…………………………………………………………………………………… 55 IV.2.3.3.Evaluation de l’activité antibactérienne……………………………………………………………. 56 IV.2.3.3.1.Identification des souches étudiées………………………………………………………………. 56 IV.2.3.3.2.Préparation des suspensions bactériennes……………………………………………………. 59 IV.2.3.3.3.Tests de l’activité antibactérienne…………………………………………………………………. 60 IV.2.3.4.Evaluation de l’activité antifongique………………………………………………………………… 61 IV.2.4.Evaluation de l’activité antioxydante…………………………………………………………………… 61 IV.2.4.1.Test au DPPH……………………………………………………………………………………………………. 61 V .Résultats et discussion…………………………………………………………………………………………… 62 V.1.Rendement d’extraction………………………………………………………………………………………….. 62 V.1.1.Les huiles essentielles…………………………………………………………………………………………… 62 V.1.2.Les extraits organiques…………………………………………………………………………………………. 63 V.2.Résultat d’analyse des huiles essentielles………………………………………………………………… 64 V.2.1.La plante Pulicaria arabica……………………………………………………………………………………. 64 V.3.Résultats du test du pouvoir antimicrobien……………………………………………………………… 68 V.3.1.L’ activité antibactérienne……………………………………………………………………………………. 68 V.3.1.1.Identification des souches étudiées…………………………………………………………………… 68 V.3.1.1.1.Caractères morphologiques et culturaux………………………………………………………… 68 V.3.1.1.2.Identification biochimiques……………………………………………………………………………. 69 V.3.1.1.3.Antibiogramme………………………………………………………………………………………………. 71 V.3.1.2.Tests de l’activité antibactérienne…………………………………………………………………….. 74 V.3.1.2.1.Control négatif……………………………………………………………………………………………….. 74 V.3.1.2.2.Les extraits de Rhanterium adpressum…………………………………………………………… 75 V.3.1.2.3.Les extraits de Pulicaria arabica……………………………………………………………………… 83 V.3 .2.L’ activité antifongique………………………………………………………………………………………… 94 V.4.Résultats du test du pouvoir antioxydant………………………………………………………………… 96 V.4.1.Acide ascorbique………………………………………………………………………………………………….. 97 V.4.2.Les extraits de la plante Rhanterium adpressum…………………………………………………… 98 V.4.3.Les extraits de la plante Pulicaria arabica…………………………………………………………….. 102 CONCLUSION……………………………………………………………………………………………………………. 108 REFERENCES BIBLIOGRAPHYQUE……………………………………………………………………………… 110 ANNEXE GLOSSAIRE RESUME REMERCIEMENTS Je remercie DIEU
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