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Le Bioraffinage De Myrrhis Odorata, Tussilago Farfara Et Calamintha Grandiflora Pour La Production D'aromes Et D'antioxydants

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Le Bioraffinage De Myrrhis Odorata, Tussilago Farfara Et Calamintha Grandiflora Pour La Production D'aromes Et D'antioxydants En vue de l'obtention du DOCTORAT DE L'UNIVERSITÉ DE TOULOUSE Délivré par : Institut National Polytechnique de Toulouse (INP Toulouse) Discipline ou spécialité : Sciences des Agroressources Présentée et soutenue par : Mme DIANA DOBRAVALSKYTE le lundi 25 novembre 2013 Titre : LE BIORAFFINAGE DE MYRRHIS ODORATA, TUSSILAGO FARFARA ET CALAMINTHA GRANDIFLORA POUR LA PRODUCTION D'AROMES ET D'ANTIOXYDANTS. Ecole doctorale : Sciences de la Matière (SM) Unité de recherche : Laboratoire de Chimie Agro-Industrielle (L.C.A.) Directeur(s) de Thèse : M. THIERRY TALOU M. RIMANTAS VENSKUTONIS Rapporteurs : Mme CHANTAL MENUT, UNIVERSITE MONTPELLIER 2 Mme JOLANTA LIESIENE, UNIVERSITE DE TECHNOLOGIE DE KAUNAS Membre(s) du jury : 1 M. ZIGMUNTAS BERESNEVICIUS, UNIVERSITE DE TECHNOLOGIE DE KAUNAS, Président 2 M. ALGIRDAS ZEMAITAITIS, UNIVERSITE DE TECHNOLOGIE DE KAUNAS, Membre 2 Mme GRAZINA JUODEIKIENÉ, UNIVERSITE DE TECHNOLOGIE DE KAUNAS, Membre 2 Mme ISABELLE FOURASTE, UNIVERSITE TOULOUSE 3, Membre 2 Mme RUTA GALABURDA, LATVIA UNIVERSITY OF AGRICULTURE JELGAVA, Membre 2 M. RIMANTAS VENSKUTONIS, UNIVERSITE DE TECHNOLOGIE DE KAUNAS, Membre REMERCIEMENTS Tout d’abord j’adresse mes remerciements aux différents organismes qui ont mis les moyens financiers pour l’aboutissement de cette thèse: aux gouvernements de la Lituanie et la France, ainsi que Campus France avec la bourse d'Excellence Égide Eiffel. Je remercie les Professeurs Dr. Jolanta LIESIENĖ et Dr. Chantal MENUT pour m'avoir fait l'honneur d'être les rapporteurs de ma thèse. Je remercie également les professeurs Dr. Algirdas ŽEMAITAITIS, Dr. Gražina JUODEIKIENĖ, Dr. Zigmuntas Jonas BERESNEVIČIUS, Dr. Isabelle FOURASTÉ, Dr. Audrius Sigitas MARUŠKA et Dr. Ruta GALOBURDA d’avoir participés en tant qu’examinateurs. Je remercie le Professeur Petras Rimantas VENSKUTONIS d’avoir accepté d’être mon directeur de thèse. J’exprime toute ma gratitude à mon co-directeur de thèse, Dr. Thierry TALOU, pour son encadrement et ses conseils pendant ces années de thèse. Je voudrais exprimer mes plus sincères remerciements à l'ensemble des doctorants et tout le personnel et département du Laboratoire de Chimie Agro-Industrielle et mes collègues de bureau. Également aux doctorats de Génie industriel à l’ENSIACET. En fin, je voudrais remercie à Jesus Manuel, mon mari, pour ses conseils et son soutien pendant la rédaction du manuscrit de thèse et finalement a ses amis pour leur amitié. iii iv RESUME INTRODUCTION Contexte Les différentes plantes aromatiques, médicinales et d’épices sont une immense source de ressources naturelles et durables de composés avec des propriétés bénéfiques. Elles sont utilisées, depuis la préhistoire, comme aromatisants ou préservent des aliments ou comme traitement médicaux. Actuellement, certains d'entre eux sont déjà cultivés commercialement pour utiliser ses propriétés. Néanmoins, il existe encore certaines plantes médicinales et aromatiques qui ne sont pas encore valorisées et donc elles sont dénommées comme « plantes médiévales oubliées ». Par exemple, Calamintha grandiflora, Myrrhis odorata et Tussilago farfara correspondent à ce type de plantes et lesquelles seront considérées dans ce manuscrit. Afin de revaloriser ces plantes oubliées, la bioprospection est mise en place et laquelle permet la recherche, la collection de nouveaux composés bioactifs naturels et son utilisation dans les produits pharmaceutiques et cosmétiques, par exemple. De plus, la bioprospection permet d’analyser les profils chimiques de différentes plantes et définir une liste des composés phytochimiques associés à chaque plante. Ainsi, par exemple, au travers de l’analyse d’une plante spécifique, il est possible définir les différents types antioxydants existant ainsi que sa quantité. Donc, cette plante peut utiliser comme une plante médicinale ou aromatique pour la consommation humaine afin d’absorber et neutraliser les radicaux libres ou la décomposition des peroxydes dangereux. L'apparition des plantes médicinales et aromatiques et leur composition dépendent de nombreux facteurs, mais particulièrement, sur les conditions climatiques et le type de sol. La biogenèse des composés biologiquement actifs dépend de plusieurs facteurs: l’origine de la plante, la zone géographique, son génotype, phénotype et chémotype, ainsi que les fluctuations climatiques annuelles. La plupart des composés naturels possèdent plusieurs activités biologiques: antioxydants, antimicrobiens, anti-inflammatoires et autres. En conséquence, la comparaison des espèces végétales cultivées dans des sites géographiques différents afin d'évaluer leurs propriétés biologiques, il est une tâche intéressante et importante, en particulier, pour la valorisation de ces plantes pour leur culture commerciale. v Par ailleurs, une autre tâche importante pour la valorisation est l’utilisation rationnelle des plantes dans toute sa totalité, c’est-à-dire, la valorisation de tout le produit avec le moins de déchets. Les déchets ou sous-produits obtenus lorsque la transformation ou modification de transformer des matières premières des plantes peuvent représenter une ressource potentielle des antioxydants et d'autres substances importantes. Par exemple, les déchets produits pendant la distillation des plantes aromatiques pour la récupération de l'huile essentielle peuvent constituer jusqu'à 99,5 % des matières premières lesquels sont utilisés inefficacement ou ne sont pas utilisés. En effet, il y a des preuves croissantes que les composés non volatils présents dans ces résidus peuvent posséder diverses activités, qui pourraient être appliquées dans les aliments comme antioxydants et des additifs antimicrobiens pour la prévention de diverses maladies telles que le cancer, l'athérosclérose et des difficultés neurologiques. De plus, en considérant les études récentes [1-3] sur la fraction volatile de plantes, traditionnellement appelées huiles essentielles, qui considèrent leurs propriétés antioxydantes comme des substituts potentiels pour les antioxydants synthétiques [4]. Il est possible d’utiliser (ou revaloriser) les sous-produits, qui sont obtenus après l'élimination de l'huile volatile par diverses techniques de distillation, afin d’améliorer la faisabilité commerciale de la production de composés bioactifs de différentes plantes. Dans notre recherche nous sommes intéressés par des espèces qui sont considérées comme de « plantes oubliées médiévales » et dans lesquelles les sous-produits ne sont pas encore analysés dans les études récentes. Ainsi, Calamintha grandiflora, Myrrhis odorata et Tussilago farfara, qui sont des plantes de type et qui poussent en Lituanie et au sud de la France, sont considérées comme candidat à réaliser une étude afin d’identifier les composant leurs propriétés et ses activités (antioxydants ou odorants) et définir la variation des composés bioactifs entre les différentes origines, une tâche considérée importante tant du point de vue chimique et pour l'optimisation de la production d'ingrédients fonctionnels bioactifs. Objectif et les tâches du travail Le but de ces travaux de thèse était d'évaluer les possibilités de la transformation des plantes Calamintha grandiflora, Myrrhis odorata et Tussilago farfara en arômes et ingrédients antioxydants en utilisant le concept d’agro-raffinerie. Les tâches suivantes ont été définies pour atteindre l'objectif: vi 1. Déterminer la composition d'huiles essentielles à partir de plantes aromatiques collectées en Lituanie et sud de la France et les comparer avec les données publiées antérieurement. 2. Développer de nouvelles techniques d'extraction rapide des composés volatils et les comparer avec les méthodes traditionnelles. Pour ce faire, les composés volatils des huiles essentielles de Calamintha grandiflora obtenues par la méthode d'hydrodistillation sont comparés avec ceux obtenus avec les nouvelles méthodes Artificial Crushing Finger device (CFD) et Design of Flash Aroma Dispenser (FAD). 3. Évaluer les propriétés antioxydantes des sous-produits d'hydrodistillation au travers des différents essais in vitro avec différents solvants. 4. Évaluer les propriétés antioxydantes des sous-produits d'hydrodistillation in situ (huile de tournesol) en le comparant avec l’antioxydant synthétique commercial. 5. Déterminer les composés phytochimiques des sous-produits d'hydrodistillation et évaluer les propriétés antioxydantes de composants individuels en utilisant des méthodes chromatographiques et spectrométriques. Originalité de la thèse La nouveauté scientifique suite à ces travaux de thèse a été réalisée par l'accomplissement des tâches énumérées ci-dessus: 1. Les profils chimiques des composés volatils dans les plantes sélectionnées qui poussent en Lituanie et au sud de la France ont été compares par première fois. Les différences entre les plantes de Lituanie et de France ont été observées, ainsi qu’avec des travaux précédents rapportés dans la littérature. 2. Deux nouvelles méthodes analytiques (Artificial Crushing Finger device et Design of Flash Aroma Dispenser) pour l'évaluation rapide de native huile essentielle de plantes aromatiques ont été développées et testées. 3. L'évaluation systématique et complète des composés phytochimiques dans les sous- produits d'hydrodistillation, particulièrement les composés phénoliques, dans Calamintha grandiflora, Myrrhis odorata et Tussilago farfara a été réalisée pour la première fois. 4. Résidus désodorisés de Calamintha grandiflora, Myrrhis odorata et Tussilago farfara ont été valorisés à partir des bioressources d'un antioxydant pour la première
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