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ETUDES CHIMIQUE ET TOXICOLOGIQUE DES EXTRAITS DE FRUITS D’Aframomum Angustifolium (ZINGIBERACEAE)

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ETUDES CHIMIQUE ET TOXICOLOGIQUE DES EXTRAITS DE FRUITS D’Aframomum Angustifolium (ZINGIBERACEAE) UNIVERSITE D’ANTANANARIVO FACULTE DES SCIENCES DOMAINE SCIENCES ET TECHNOLOGIE ************************ MENTION BIOCHIMIE FONDAMENTALE ET APPLIQUEE ************************************ Mémoire pour l’obtention du Diplôme de Master II Parcours : Biochimie, Biodiversité et Santé ETUDES CHIMIQUE ET TOXICOLOGIQUE DES EXTRAITS DE FRUITS D’Aframomum angustifolium (ZINGIBERACEAE) présenté par : RAKOTONIRINA Fandresena Nacya le 19 avril 2019 devant le jury composé de : - Président : Professeur RASAMIRAVAKA Tsiry - Rapporteur : Professeur RAKOTO Danielle A. Doll - Examinateurs : Docteur RAZAFIARIMANGA Zara Docteur RANDRIAMAMPIANINA Lovarintsoa Judicaël TABLE DES MATIERES REMERCIEMENTS ..................................................................................................... i GLOSSAIRE ................................................................................................................ ii LISTE DES ABREVIATIONS ................................................................................... iii LISTE DES FIGURES ................................................................................................ iv LISTE DES TABLEAUX ............................................................................................ v INTRODUCTION GENERALE ................................................................................. 1 SYNTHESE BIBLIOGRAPHIQUE ............................................................................ 4 1. GENERALITES SUR LE GENRE Aframomum ................................................. 4 2. UTILISATIONS DES Aframomum ..................................................................... 4 2.1 UTILISATIONS DANS LE MONDE ............................................................ 4 2.2 UTILISATIONS D’Aframomum angustifolium A MADAGASCAR ............ 5 2.3 REPARTITION GEOGRAPHIQUE .............................................................. 7 3. DONNEES PHARMACOLOGIQUES ................................................................ 7 4. PRINCIPAUX METABOLITES SECONDAIRES RENCONTRES.................. 8 PREMIERE PARTIE : ETUDE CHIMIQUE I. INTRODUCTION .................................................................................................. 9 II. MATERIELS ET METHODES .......................................................................... 9 II.1 MATERIELS ..................................................................................................... 9 II.1.1 Matériel végétal ......................................................................................... 9 II.1.1.1 Taxonomie ............................................................................................ 9 II.1.1.2 Description botanique de l’espèce angustifolium ................................. 9 II.1.1.3 Date et lieu de récolte ......................................................................... 10 II.1.1.4 Préparation du matériel d’étude.......................................................... 10 II.1.2 Les produits chimiques et les autres matériels..................................... 10 II.2 METHODES .................................................................................................... 10 II.2.1 Méthodes d’extraction des principes actifs .......................................... 10 II.2.1.1 Extraction à froid ................................................................................ 10 II.2.1.2 Extraction à chaud .............................................................................. 11 II.2.2 Méthodes de purification ....................................................................... 11 II.2.2.1 Traitement par la chaleur .................................................................... 11 II.2.2.1.1 Principe ....................................................................................... 11 II.2.2.1.2 Mode opératoire .......................................................................... 11 II.2.2.2 Fractionnement par le n-butanol ......................................................... 11 II.2.2.2.1 Principe ....................................................................................... 11 II.2.2.2.2 Mode opératoire .......................................................................... 12 II.2.3 Méthodes analytiques ............................................................................. 12 II.2.3.1 Chromatographie sur couche mince ................................................... 12 II.2.3.1.1 Principe ....................................................................................... 12 II.2.3.1.2 Mode opératoire .......................................................................... 12 II.2.3.2 Méthodes de criblage phytochimique ................................................. 13 II.2.3.2.1 Principe ....................................................................................... 13 II.2.3.2.2 Test sur l’extrait acide ................................................................ 13 II.2.3.2.3 Tests sur l’extrait hydroéthanolique .......................................... 14 a) Détection des flavonoïdes ....................................................................... 14 b) Détection des leucoanthocyanes ............................................................. 14 II.2.3.2.4 Tests sur l’extrait chloroformique ............................................. 14 a) Détection des stéroïdes et triterpènes ..................................................... 14 b) Détection des stérols insaturés ............................................................... 14 II.2.3.2.5 Tests sur l’extrait aqueux ........................................................... 14 a) Détection des anthraquinones ................................................................. 15 b) Détection des iridoïdes ............................................................................ 15 c) Détection des saponines .......................................................................... 15 d) Détection des tanins et autres polyphénols ............................................. 15 e) Détection des désoxyoses ........................................................................ 15 II.2.4 Méthode de concentration ...................................................................... 16 II.2.5 Calcul de rendement ............................................................................... 16 III. RESULTATS ..................................................................................................... 16 III.1 EXTRAITS OBTENUS ................................................................................. 16 III.2 PURIFICATION ............................................................................................ 17 III.2.1 Traitement par lachaleur ..................................................................... 17 III.2.2 Fractionnement par le n-butanol ......................................................... 17 III.3 DEGRE D’HOMOGENEITE DES DIFFERENTS PRODUITS .................. 18 III.4 CARACTERISATION CHIMIQUE .............................................................. 19 III.4.1 Propriétés physico-chimiques .............................................................. 19 III.4.2 Nature chimique .................................................................................... 19 III.5 RENDEMENTS ............................................................................................. 20 IV. DISCUSSION ET CONCLUSIONS ................................................................ 21 DEUXIEME PARTIE : ETUDE TOXICOLOGIQUE I. INTRODUCTION ................................................................................................ 22 II. MATERIELS ET METHODES ........................................................................ 22 II.1 MATERIELS ................................................................................................... 22 II.1.1 Les animaux d’expérimentation ............................................................ 22 II.1.1.1 Animaux à sang chaud : les souris ..................................................... 22 II.1.1.2 Animaux à sang froid : les larves de moustique ................................. 22 II.1.2 Les végétaux d’expérimentation ............................................................ 22 II.1.3 Matériels de microbiologie ..................................................................... 23 II.1.3.1 Les germes utilisés ............................................................................. 23 II.1.3.2 Le milieu de culture ............................................................................ 23 II.1.3.3 Les disques pour antibiogramme ........................................................ 23 II.2 METHODES .................................................................................................... 24 II.2.1 Méthodes d’étude des effets sur les animaux ....................................... 24 II.2.1.1 Tests sur animaux à sang chaud ......................................................... 24 II.2.1.1.1 Estimation de la toxicité ............................................................. 24 II.2.1.1.2 Détermination de la DL50 (24h) ................................................ 24 II.2.1.2 Test sur les animaux à sang froid ....................................................... 25 II.2.1.2.1 Principe ....................................................................................... 25 II.2.1.2.2 Mode opératoire .......................................................................... 25 II.2.2
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