Dissertations / Theses on the topic 'Hydrodistillation'

Garlic essential oil possesses several desired pharmacological effects. Microwave-assisted hydrodistillation (MAHD) was adopted as to reduce the extraction period and improve extraction yield. The antimicrobial properties of the extracted essential oils were analysed and a rigorous fundamental model was developed to represent the process. The results showed that the MAHD combined with water-ethanol solvent can increase the extraction rate, shorten the extraction period and maintain the antimicrobial property of the garlic oil.

ABSTRACT SOLVENT-FREE MICROWAVE EXTRACTION AND MICROWAVEASSISTED HYDRODISTILLATION OF ESSENTIAL OILS FROM SPICES Bayramoglu, Beste M.S., Department of Food Engineering Supervisor: Assoc. Prof. Dr. Serpil Sahin Co-Supervisor: Assoc. Prof. Dr. S. Gü
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September 2007, 130 pages The undesirable effects of conventional methods generated the need for economical and safe techniques in the extraction of essential oils. Microwave-assisted hydrodistillation (MAHD) and solvent-free microwave extraction (SFME) are recently developed techniques, which are thought to overcome this problem. Oregano (Origanum vulgare L.), laurel (Laurus nobilis L.) and rosemary (Rosmarinus officinalis L.) were chosen in this study since they have high antimicrobial and antioxidant effects and are widely grown and consumed in Turkey. The objectives of this study were to examine the applicability of SFME in the extraction of essential oils from oregano and laurel, and MAHD in the extraction of rosemary essential oil. The effects of microwave power and extraction time on the yield, composition, and other quality parameters of the extracts were also investigated. Hydrodistillation was performed as control. SFME offered significantly higher essential oil yields (0.054 mL oil/g oregano) from oregano as compared to hydrodistillation (0.048 mL oil/g oregano). Conventional process time was reduced by 80%. Main aroma compound was thymol (650-750 mg thymol/mL oil). For laurel, no significant differences were obtained in yields (about 0.022 mL oil/g laurel) obtained by SFME and hydrodistillation. Process time was reduced by 55-60%. Main aroma compound was 1,8-cineole (630-730 mg 1,8-cineole/mL oil). In the case of rosemary, no significant differences were obtained in yields (about 0.026 mL oil/g rosemary) obtained by MAHD at 622 W and hydrodistillation. The process time was reduced by about 65%. Main aroma compounds were 1,8-cineole (430-500 mg 1,8-cineole/mL oil) and camphor (150-210 mg camphor/mL oil). Keywords: Solvent-free microwave extraction (SFME), Microwave-assisted hydrodistillation (MAHD), Origanum vulgare L., Laurus nobilis L., Rosmarinus officinalis L.

Ce travail concerne l'extraction de substances naturelles d'origine végétale en utilisant des procédés « verts » tels que l'extraction au CO2 supercritique (CO2-SC, de 90 à 1000 bar) et l'extraction à l'eau sub-critique (SWE). Ces procédés sont une alternative à l'hydrodistillation (HD) et l'extraction par solvant (SE) traditionnellement utilisés dans l'industrie des produits naturels. La majorité du travail a été réalisée sur l'espèce Tetraclinis articulata où les performances des procédés, CO2-SC , HD, SWE et SE, ont été optimisées puis comparées en termes de rendement, composition chimique, durée…, et en particulier activité antioxydante des extraits (évaluée par les tests ABTS+ and DPPH). La plus forte activité antioxydente a été mise en évidence pour les extraits CO2-SC, ceci étant très probablement dû à une moindre dégradation thermique, comme l'a indiqué la comparaison des compositions chimiques. La CO2-SC basse pression (90 bar) a permis d'obtenir un extrait de bonne qualité et, pour ce cas, une modélisation a été proposée pour comprendre et identifier le mécanisme limitant qui s'est avéré être l'équilibre solide-fluide. La modélisation de l'hydrodistillation a également été proposée, basée sur le modèle de Sovová . Ensuite, l'approche a été étendue à d'autres plantes : Eucalyptus cinerea, Eucalyptus Camaldulensis, Cypres sempervirens et callitris. ABSTRACT : This work has dealt with extraction of natural substances from plants using « green » processes such as supercritical CO2 extraction (CO2-SC from 90 to 1000 bar) and extraction with subcritical water (SWE). These processes are an alternative to hydrodistillation (HD) and solvent extraction (SE) traditionally used in the natural products industry. Main part of the work was done on the species Tetraclinis articulata, and performance of the different processes, CO2-SC, HD,SWE and SE, were optimized and compared in terms of yield, chemical composition, duration ... and especially antioxidant activity of extracts (assessed by the ABTS+ and DPPH tests). CO2-SC extracts exhibited the strongest antioxidant activity and comparison of chemical compositions of the different extracts indicated that this is very probably due to lower thermal degradation of active molecules. For low pressure CO2-SC (90 bar), a model was proposed to understand and identify the limiting mechanism that proved to be solid-fluid equilibrium. For hydrodistillation, modelling, based on the Sovová 's model was also proposed. Then, the approach was extended to other plants: Eucalyptus cinerea and Eucalyptus camaldulensis, Cypres sempervirens and Callitris.

>Magister Scientiae - MSc
Pelargonium capitatum is a plant that is popular in essential oils. The isolated essential oil constituents from the flowers and leaves of the plant were obtained by hydrodistillation followed by GC-MS analysis. The following essential constituents were obtained in abundance from flowers and leaves 8, 11-octadecadienoic acid and citronellol, caryophyllen, α-cubebene, copaene, azulene, pentacosane, 9, 12- octadecadienoic acid. Crude extracts from Pelargonium capitatum were fractionated through various chromatographic techniques in order to achieve satisfactory separations. Four compounds were characterized and the structural elucidation were unambiguously confirmed by spectroscopic methods including one and two-dimensional nuclear magnetic resonance and high resolution mass spectroscopy. The compounds were isolated from the ethyl acetate (2) and hexane (2) extracts, respectively. The four isolates were identified as quercetin, quercetin xylopyranoside, β-sitosterol and undecaprenol. The isolate and the different extracts were tested for cytotoxicity using brine shrimp lethality test. The results showed that quercetin and the extracts were active with hexane extract showed the highest cytotoxicity level of 1.5μg/ml. To the best of our knowledge quercetin xylopyranoside, β-sitesterol and undecaprenol compounds are reported for the first time from Pelargonium capitatum.

Les exsudats des plantes sont des substances naturelles complexes. Ils sont utilisés par plusieurs populations d’Afrique et trouvent de nombreuses applications : en thérapeutique, en pharmacie galénique, en parfumerie, etc. Au Congo, une grande majorité de ces substances est encore mal connue. Ce travail de thèse a consisté à inventorier les plantes à exsudats du Congo et à mettre au point une méthodologie d’analyse physico-chimique permettant la caractérisation des exsudats d’arbres d’origine botanique certifiée.La méthodologie d’inventaire adoptée nous a permis de recenser 135 plantes à exsudats appartenant à 87 genres. Il en ressort de cette étude que les exsudats des plantes inventoriées ont multiples usages (glu, insectifuge, allume-feu, antidouleur, etc.).La stratégie mise en place pour leur caractérisation fait appel à plusieurs techniques analytiques (SPIR, SMIR, CPG-SM, GCxGC-MS) et de traitement d’échantillon (Hs-SPME, hydrodistillation, extraction au Soxhlet, silylation, méthylation, pyrolyse). Dans le cadre de ce travail deux exsudats ont été caractérisés. L’exsudat de D. edulis (G. Don) K.J. Lam qui est constitué de monoterpènes hydrocarbonés et de triterpènes, dont les majeurs sont : 3-épi-α et -β-amyrine. L’exsudat de G. demeusei (Harms) J. Léonard dont la fraction volatile est constituée essentiellement de sesquiterpènes et la fraction peu-volatile d’acides diterpéniques de type labdane (acide éperuique, cativique, copalique, ozique, etc.).Pour exploiter l’efficacité de séparation élevée de la GCxGC-MS, nous nous sommes confronté aux problèmes de la non-répétabilité des cartes chromatographiques obtenues en 2D. Nous avons alors mis en place une méthodologie permettant de vérifier l’épaisseur de film et l’homogénéité des dépôts de phases stationnaires afin de sélectionner des colonnes courtes identiques pour la GCxGC. Cependant, les résultats préliminaires ont montré que même avec des colonnes de deuxième dimension identiques, on pouvait avoir des différences au niveau de carte 2D
Plant exudates are natural complex substances. They are used by several populations in Africa and have many applications: in therapeutics, in galenic pharmacy, in perfumery, etc. In Congo, a great majority of these substances are not yet well known. This thesis work aimed to make an inventory of plant exudates from Congo and to develop a methodology for performing physico-chemical analysis which may enable characterization of exudates from botanical certified trees.Through the methodology that we developed, 135 plant exudates belonging 87 genera were registered. It was observed in this study that these exudates are used for many purposes (glue, insect repellent, making fire, anti-pain, etc).The strategy implemented for characterizing the exudates included several analytical (SNIR, SMIR, GC-MS, GCxGC-MS) and sample processing (Hs-SPME, hydrodistillation, extraction using the Soxhlet, silylation, methylation pyrolyse) techniques. With regard to this work, two exudates were characterized. The exudates from D. edulis (G. Don) K.J. Lam which contained monoterpene hydrocarbons and triterpenes, with the major constituents being: 3-epi-α and -β-amyrine. The exudates from G. Demeusei (Harms) J. Léonard was found to have the sesquiterpenes as major constituents of the volatile fraction and the labdane-type diterpenes (eperuique acide, cativique acid, copalique acid, ozique acid, etc) as major constituents of the poor-volatile fraction.To explore the efficacy of elevated separation of the GCxGC-MS, we faced some problems related to the non-repeatability of chromatographic cards obtained in 2D. Therefore, we put in place a methodology which enables us to verify the film thickness and homogeneity of layers of stationary phases in order to select identical short columns for GCxGC. However, the preliminary results have shown that even using second identical dimension columns, we could observed differences in 2D cards

La recherche de nouveaux debouches pour les co-produits de culture du tournesol, a ete motivee par les tonnages eleves de residus generes. Dans un premier temps, une etude approfondie des caracteristiques physico-chimiques des differentes parties de la plante, debarrassee de ses graines, nous a permis de cibler deux produits interessants : une huile essentielle fortement odorante utilisable en parfumerie, et des pectines, connues pour leur teneur elevee dans le capitule, mais egalement presentes dans la moelle de la tige. Un approfondissement de cette etude a permis d'identifier les principaux constituants de huile essentielle, jusqu'a present inconnue. Parallelement, l'analyse de l'evolution des pectines de capitule au cours de la croissance de la plante nous a amene a relier leur faible degre de methylation (deja connu), a l'activite accrue des pectines methylesterases (pme) en fin de maturation de la plante. Ceci a conduit a envisager une strategie transgenique, ayant pour but de controler l'action des pme afin d'obtenir des pectines de degre de methylation superieur, utilisables dans le domaine des gelifiants. Une autre voie de valorisation des pectines a egalement ete etudiee a travers leur modification chimique, par esterification, avec des alcools a longue chaine. Cette etude a abouti a la synthese, sous activation micro-ondes, de molecules tensioactives dont les proprietes pourront offrir de nouvelles opportunites dans des domaines tels que les detergents ou les peintures. Enfin, dans une derniere partie, l'ensemble du procede de valorisation est aborde, incluant : les problemes lies a l'organisation de la recolte, les procedes d'obtention des extraits a l'echelle pilote, et la fabrication, suivie de la caracterisation mecanique, de materiaux type carton ondule, ou de materiaux faible densite a base de moelle destines aux domaines du calage et de l'emballage. En conclusion, un schema global de valorisation des co-produits est propose.

La présence d’huiles essentielles dans la peau des fruits et des feuilles constitue une spécificité des agrumes tels que l’orange et la bigarade. Cette huile essentielle contient de nombreux composés, variables en quantité et en nombre selon l’espèce ou la variété, et dont certains possèdent des propriétés aromatiques très appréciées par les consommateurs et par l’industrie agroalimentaire et des cosmétiques et des parfums. La quantité des composés aromatiques peut varier en fonction de nombreux facteurs liés aux techniques culturales, à l’environnement (climat, sol), à la maturité des fruits, aux méthodes de séchage de la peau et à la génétique (variétés et hybrides). Cette variation de composition de l’huile peut provoquer une modification de l’arôme et donc changer les caractéristiques du fruit ou d’un produit transformé. La thèse avait donc comme objectif d’évaluer l’impact sur la composition chimique et le profil aromatique : du porte-greffe, de la ploïdie, du stade de développement du fruit, de la variabilité, du séchage et de la conservation de la peau des fruits et la génétique liée à la mutation (diversité variétale) et celle produite par l’hybridation. Les huiles essentielles ont été extraites par hydrodistillation, leur composition déterminée par chromatographie en phase gazeuse couplée avec un détecteur à ionisation de flamme et avec un spectromètre de masse. En parallèle leurs arômes ont été caractérisés par différentes analyses sensorielles. Des marqueurs génétiques ciblant des microsatellites et le polymorphisme nucléotidique ont été utilisés pour évaluer la diversité génétique de la collection variétale des deux espèces d’agrumes et construire une carte génétique de l’oranger via une approche de génotypage par séquençage pour localiser les locus de caractères quantitatifs des composés aromatiques.La composition chimique de l’huile essentielle de zestes des deux agrumes est dominée par le limonène qui représente plus de 90% de l’huile essentielle ce qui gêne l’analyse de la variabilité des autres composés. La composition chimique et le profil aromatique des huiles essentielles sont gouvernés par plusieurs facteurs. L’hybridation est évidemment le facteur le plus perturbant car l’oranger est un hybride interspécifique donc très hétérozygote et très peu d’hybrides clémentinier/oranger reproduisent un profil aromatique semblable à l’orange. A l’opposé, la diversité variétale surtout basée sur la mutation n’engendre que très peu de variation tant aromatique que chimique. Néanmoins, chez les bigaradiers des profils atypiques ont été détectés issus d’hybridations identifiées comme différentes de la phylogénie des bigaradiers classiques. Le stade de développement du fruit a aussi une forte influence mais seulement lorsque les fruits sont petits et immatures (juillet – septembre). Le porte-greffe ainsi que son niveau de ploïdie et celui du scion n’ont qu’une faible influence sur la composition et le profil aromatique. En post-récolte, la méthode de séchage est très influente en induisant sous l’action de la chaleur la dégradation de composés de la famille des monoterpènes oxygénés et des aldéhydes aliphatiques ce qui entraine la détérioration du profil aromatique. En revanche, tous les facteurs étudiés modifient de manière importante le rendement en huile. La cartographie génétique a localisé vingt locus de caractères quantitatifs de quinze composés et du rendement en huile essentielle dont certains positionnés dans les mêmes groupes de liaison.Au cours de cette thèse, les principaux facteurs d’instabilité de l’huile essentielle d’orange et de bigarade ont été quantifiés et des suggestions ont été faites quant aux travaux complémentaires à réaliser pour mieux contrôler la qualité des arômes. Des recommandations de traitements à appliquer sur le séchage des peaux d’orange et de bigarade pour préserver au mieux leur qualité aromatique ont été proposées
The presence of essential oils in the fruit skin and leaves is a specificity of citrus fruits such as orange and sour orange. This essential oil contains numerous compounds, varying in quantity and number according to the species or variety, and some of which have aromatic properties highly appreciated by consumers and by the agri-food, cosmetics and perfume industries. The quantity of aromatic compounds can vary according to many factors related to cultivation techniques, environment (climate, soil), fruit maturity, skin drying methods and genetics (mutants and hybrids). This variation in the essential oil composition can cause a modification of the aroma and thus change the characteristics of the fruit or of a processed product. The objective of the thesis was therefore to evaluate the impact on the chemical composition and aroma profile of rootstock, ploidy, stage of fruit development, variability, drying and preservation of the fruit skin, and genetics related to mutation (varietal diversity) and that produced by hybridization (progeny). The essential oils were extracted by hydrodistillation, their composition determined by gas chromatography coupled with a flame ionization detector and a mass spectrometer. In parallel, their aromas were characterized by different sensory analyses. Genetic markers such as microsatellites (SSR) and nucleotide polymorphism (SNP) were used to assess the genetic diversity of the varietal collection of sweet and sour oranges and to build a genetic map of orange via a genotyping sequencing approach (GBS) to locate the quantitative trait loci (QTLs) of the aromatic compounds.The chemical composition of the peel essential oil (PEO) of both citrus fruits is dominated by limonene representing more than 90% of the total hindering the analysis of the variability of other compounds. The chemical composition and aromatic profile of essential oils are modified by several factors. Hybridization is obviously the most disturbing factor because the orange tree is an interspecific hybrid and therefore very heterozygous, and very few clementine/orange hybrids reproduce an aromatic profile similar to that of orange. On the other hand, varietal diversity, mainly based on mutation, generates very little variation, either aromatic or chemical. Nevertheless, atypical profiles of few sour orange cultivars have been identified as hybrids originated from a different cross than the regular genotypes. The stage of fruit development has also a strong influence but only when the fruits are small and immature (July - September). The rootstock and the ploidy level have only a weak influence on the composition and aromatic profile of orange PEO. In post-harvest, the drying method induces, under the action of heat, the disappearance of compounds of the family of oxygenated monoterpenes and aliphatic aldehydes, which leads to the deterioration of the aromatic profile. On the other hand, all the factors studied significantly modify the oil yield. Genetic mapping located twenty quantitative trait loci of fifteen compounds and essential oil yield, some of which were positioned in the same linkage groups.During the course of this thesis, the main factors of instability in orange and sour orange essential oil were quantified and suggestions were made for further work to better control flavor quality. Recommendations of treatments to be applied on orange and sour orange peels drying to preserve their aromatic quality were proposed

A celulose das árvores do gênero Eucalyptus é um dos principais elementos de interesse econômico desse vegetal. Suas folhas, apesar de conter óleo essencial (OE), caracterizam-se como um subproduto da indústria de processamento de papel e celulose. Assim, pesquisas com relação à recuperação dessa biomassa tornam-se importantes, principalmente, do ponto de vista ambiental e econômico. Tradicionalmente, a extração de OEs ocorre pelo processo de hidrodestilação (HD), o qual necessita de longos tempos de extração. Visando superar essa restrição, a tecnologia de extração assistida por micro-ondas (Microwave Assisted Extraction - MAE) vem sendo desenvolvida e aplicada. O objetivo principal deste estudo foi avaliar o processo de extração de OE de folhas de eucalipto, empregando os métodos de extração HD e MAE. Para esse processo em escala de bancada, desenvolveu-se um aparato de extração por meio da adaptação de um forno micro-ondas doméstico. Inicialmente, analisou-se o rendimento de OE das matrizes de subprodutos de folhas de eucalipto das espécies Eucalyptus saligna, E. urohpylla e E. urophylla x globulus geradas a partir de uma indústria de celulose. Os resultados apontaram que a espécie E. urophylla x globulus apresentou maior teor de OE (2,16±0,02%), sendo a espécie escolhida para a aplicação da tecnologia MAE. Na segunda etapa do estudo, fez-se uma avaliação dos efeitos dos fatores do processo MAE, avaliando os fatores razão sólido:solvente (1:1; 1:1,5 e 1:2), potência do micro-ondas (680, 850 e 1.020 W) e tempo de extração total (20, 40, 60 min) no rendimento de OE. Desse modo, as condições ideais do processo foram determinadas como: razão sólido:solvente de 1:2, potência de 680 W e tempo de extração total de 60 minutos. Para essas condições, realizou-se o estudo cinético e modelagem matemática com a avaliação dos modelos de primeira ordem, segunda ordem, Peleg e Patricelli. No estudo cinético, o tempo total do processo MAE para a obtenção do rendimento de 1,8±0,1% foi apenas 60 minutos, tempo 57% inferior ao observado no método HD para igual rendimento (140 minutos). Dentre os modelos cinéticos estudados, o modelo de Patricelli foi o que apresentou melhor ajuste aos dados experimentais da extração HD (R² igual 0,9904 e RMSE igual 0,0016) e da extração MAE (R² igual 0,9962 e RMSE igual 0,0006). Por fim, as análises energéticas e de impacto ambiental também indicaram o método MAE como uma tecnologia mais ambientalmente amigável do que a HD, tornando-se atrativa para o setor industrial. Com base nos resultados obtidos, pode-se concluir que a extração de OE da matriz estudada proporcionou a reutilização de um subproduto industrial, sugerindo uma alternativa de exploração e agregando valor ao mesmo.
The cellulose from trees of the genus Eucalyptus is the main industrial product of this species. Its leaves, although containing essential oil (EO), are characterized as a by-product from the pulp and paper processing industry. Thus, researches about the recovery of this biomass are important, mainly from the environmental and ecological point of view. Traditionally, the extraction of EO occurs by hydrodistillation (HD), process that requires long extraction times. In order to overcome this restriction, microwave assisted extraction (MAE) has been developed and applied. The main objective of this study was to evaluate the extraction process of eucalyptus EO, using the HD and MAE methods. For this laboratory-scale process, an extraction apparatus was developed through the adaption of a domestic microwave oven. Initially, the yield of EO by-products of eucalyptus leaves of the species Eucalyptus saligna, E. urophylla and E. urophylla x globulus from a cellulose industry were analyzed. The results showed that the species E. urophylla x globulus presented the highest EO content (2.16 ± 0.02%), being the species chosen for the application of the MAE technology. In the second stage of the study, an evaluation of the effects of MAE process factors was performed, analyzing the factors ratio solid:solvent (1:1, 1:1,5 and 1:2), microwave power (680, 850 and 1.020 W) and total extraction time (20, 40, 60 min) in the EO yield. Thus, the ideal process conditions were determined as: ratio solid:solvent of 1:2, power of 680 W and total extraction time of 60 minutes. For these conditions, a kinetic study and a mathematical modeling were performed evaluating the first-order, second-order, Peleg and Patricelli models. In the kinetic study, the total time of the MAE process to obtain the yield of 1.8 ± 0.1% was only 60 minutes, 57% lower than the HD method for the same yield (140 min). Among the kinetic models studied, the Patricelli model presented the best fit to the experimental data of the HD extraction (R² equal to 0.9904 and RMSE equal to 0.0016) and MAE extraction (R² equal to 0.9962 and RMSE equal to 0.0006). Finally, the energy and environmental impact analyzes also indicated the MAE method more environmentally friendly than HD, being attractive for the industrial sector. Based on the results, it is possible to conclude that the extraction of EO from the studied matrix provided the reuse of an industrial by-product, suggesting an exploration alternative adding value to this residue.

Les arbres du genre Aquilaria Lam. (Thymelaeaceae), en réaction à un stress mécanique, produisent, en interactions avec des micro-organismes, un bois transformé par sa forte teneur en composés secondaires : l’agarwood (AW). Il est apprécié en parfumerie pour son odeur boisée. Face à la forte demande mondiale, des agriculteurs ont réalisé des plantations expérimentales d’Aquilaria en Guyane (projet Aquil@Guyane) afin de produire une huile essentielle de composition chimique contrôlée, respectueuse de la biologie de l’arbre et de l’environnement. Dans ce contexte, nous avons produit des connaissances académiques pour contribuer au développement de la production d’AW d’Aquilaria crassna en Guyane. Après caractérisation des communautés fongiques présentes dans les arbres d’A. crassna en Guyane, révélant le rôle des Basidiomycètes dans la production d’AW, deux méthodes d’induction d’AW ont été comparées : 1- levée d’écorce et application du sol de la plantation sur la blessure ; 2- inoculation de souches guyanaises de Basidiomycètes. Cette dernière est la plus efficace pour obtenir un AW de qualité proche d’un AW de référence. La Spectrométrie Proche Infrarouge (SPIR) a été utilisée pour différencier le bois sain de l’AW et différentes compositions chimiques d’AW. Les résultats obtenus sont cohérents avec ceux de l’analyse biochimique après hydrodistillation : utiliser la SPIR pour trier et contrôler la qualité de l’AW est envisageable. Une méthode de culture de cals d’Aquilaria a été mise au point pour comprendre les mécanismes moléculaires de la production d’AW. Les résultats indiquent une production de terpènes par les cals d’Aquilaria inoculés par des souches fongiques
Trees of the genus Aquilaria Lam. (Thymelaeaceae) in response to mechanical stress produce, in interaction with microorganisms, a wood transformed by its high content of secondary compounds: agarwood (AW). It is appreciated in perfumery for its woody fragrance. Faced with strong global demand, farmers have carried out experimental plantations of Aquilaria in French Guiana (Aquil@Guyane project) in order to produce an essential oil with a controlled chemical composition, respectful of the biology of the tree and the environment. In this context, we have produced academic knowledge to contribute to the development of AW production of Aquilaria crassna in French Guiana. After characterization of the fungal communities present in A. crassna trees in French Guiana, revealing the role of Basidiomycetes in the production of AW, two methods of induction of AW were compared: 1- lifting of bark and application of the soil of the plantation on the wound; 2- inoculation of Guyanese strains of Basidiomycetes. The latter is the most effective for obtaining a quality AW close to a reference AW. Near Infrared Spectrometry (NIRS) has been used to differentiate healthy wood from AW and different chemical compositions of AW. The results obtained are consistent with those of the biochemical analysis after hydrodistillation: using NIRS to sort and control the quality of AW is possible. An Aquilaria callus culture method has been developed to understand the molecular mechanisms of AW production. The results indicate the production of terpenes by Aquilaria callus inoculated with fungal strains

Conselho Nacional de Desenvolvimento Científico e Tecnológico
Brazil is one of the largest producers of fruit, guava with the culture of great importance in the diet of the population by having large amounts of vitamins, and carbohydrates and carotenoids. The postharvest of guava can be regarded as a problem for the broader participation of Sergipe in the marketing of this fruit in other parts of the country. The alternative control of fungal diseases plays an important role in the search for the most demanding consumer markets for healthy products and known origin. Thus the objective of this study was to develop and refine a technique that allows the storage of guavas for prolonged periods without the use of agrochemicals and excellent post-harvest quality in order to reduce costs and increase food security. Through the process of hydrodistillation essential oil was obtained from seeds of Shinus terebinthifolius from seeds and leaves and it was verified the oil yield in different periods of distillation and a period of 2.5 h distillation recommended for this plant. This had its chemical composition determined by analysis by gas chromatography, the components found in greater quantity in the oil were: ρ-Menth-1-en-9-ol, α-Thujene, β-pinene, Camphene, α - Fenchene , Terpinen-4-ol acetate, Bornyl Acetate, Cariophilene, Terpinen-4-ol, α-Terpineol, Germacrene - D, δ-cadinene, Hedycariol, α - Gurjunene, α-eudesmol, β-eudesmol. Petri dishes containing PDA culture medium supplemented with different concentrations of essential oil of S. terebinthifolius were inoculated with mycelial structures of the fungus Colletotrichum gloeosporioides for determining the lowest inhibitory concentration (MIC) of the oil on the mold where there is an efficiency of approximately 47% for controlling fungus at concentrations of 2%, 3%, 4%, 5% oil in the culture medium, as recommended concentration of 2% of essential oil in vivo tests. Guava fruit harvested at mature were inoculated with the fungus that causes anthracnose and treated with essential oil of aroeira da praia 3% being placed in low temperature (15 °C) and temperature (25 °C) in order to assess the period of storage and marketing guava and the development of the fungus against the essential oil. There was loss of weight in fruit and little variation in the amounts of soluble solids during the storage period. The guavas were already significant loss of firmness on the 5th day of storage. Significant change in the peel and pulp of guava during storage was an expected behavior for this variety of guavas. The essential oil delayed the appearance of characteristics of anthracnose lesions 3 days compared with the fungicide used in the experiment. It was demonstrated the accumulation of proline in the guavas during the post-harvest, indicating that the accumulation of proline may be related to weight loss and the development of anthracnose in guavas.
O Brasil é um dos maiores produtores mundiais de frutas, tendo a cultura da goiabeira grande importância na alimentação da população por apresentar grandes quantidades de vitaminas, além de carboidratos e carotenóides. A pós-colheita da goiaba pode ser considerada como um problema para a ampliação da participação de Sergipe na comercialização desta fruta em outras regiões do país. O controle alternativo de doenças fúngicas assume papel importante na busca de mercados consumidores mais exigentes por produtos saudáveis e de procedência conhecida. Desta forma o objetivo deste trabalho foi desenvolver e aprimorar uma técnica que possibilite o armazenamento de goiabas por períodos prolongados, sem o uso de agroquímicos e com excelente qualidade pós-colheita, de forma a reduzir os custos e aumentar a segurança alimentar. Pelo processo de hidrodestilação foi obtido o óleo essencial de sementes de aroeira da praia sendo verificado o rendimento do óleo em diversos períodos de destilação sendo o período de 2,5h de destilação o recomendado para essa planta. Esse que teve a sua composição química determinada através de análise em cromatógrafo gasoso. Os componentes encontrados em maior quantidade no óleo foram: ρ-Menth-1-em-9-ol, α-Thujene, β-Pinene, Camphene, α Fenchene, Terpinen-4-ol acetate, Bornyl Acetate, Cariophilene, Terpinen-4-ol , α-Terpineol, Germacrene D, δ-Cadinene, Hedycariol, α Gurjunene, α-Eudesmol, β-Eudesmol. Em placas de Petri contendo meio de cultura BDA acrescido de diversas concentrações do óleo essencial de aroeira da praia foram depositadas estruturas miceliais do fungo Colletotrichum gloeosporioides para a determinação da menor concentração inibitória (MCI) do óleo, onde se verificarou uma inibição de aproximadamente 47% no desenvolvimento do fungo nas concentrações de 2%, 3%, 4%, 5% do óleo no meio de cultura, sendo recomendada a concentração de 2% de óleo essencial para testes in vivo. Frutos de goiabeira colhidos fisiologicamente maduros foram inoculados com o fungo causador da antracnose e tratados com óleo essencial de aroeira da praia 3% sendo acondicionados em baixa temperatura (15 °C) e temperatura ambiente (25 °C) com o intuito de avaliar o período de armazenamento e comercialização da goiaba e também o desenvolvimento do fungo frente o óleo essencial. Verificou-se perda de massa fresca nos frutos e pouca variação nos valores de sólidos solúveis durante todo o período de armazenamento. As goiabas apresentaram perda de firmeza significativa já no 5° dia de armazenamento. Houve alteração significativa na coloração da casca e da polpa das goiabas durante o armazenamento, comportamento já esperado para essa variedade de goiaba. O óleo essencial retardou o surgimento de lesões características da antracnose em 3 dias quando comparado com o fungicida utilizado no experimento. Foi comprovado o acúmulo de prolina nas goiabas durante o período de pós-colheita, o que pode ter relação com a perda de massa fresca e com o desenvolvimento da antracnose em goiabas.

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Les huiles essentielles et les arômes des plantes constituent un réel potentiel pour l’industrie dans le but de substituer aux composés synthétiques ayant des effets néfastes sur la santé et l’environnement. Afin de contribuer aux principes de la chimie verte, cette étude porte sur l’éco-extraction et la valorisation des extraits naturels des plantes et le développement d’une nouvelle technologie «verte» pour l’extraction des composés aromatiques naturels. La première partie de ce manuscrit mets en évidence les propriétés biologiques des huiles essentielles et des arômes et l’importance de leurs applications dans différents domaines surtout le domaine agroalimentaire. Dans la deuxième partie, les études sur les propriétés biologiques des huiles essentielles des plantes des Alliacées montrent leurs bonnes activités antioxydantes et antimicrobiennes. Ces résultats encourageants ont permis dans la troisième partie de valoriser les sous-produits d’oignon issus de la turbo hydrodistillation et qui sont considérés normalement comme déchets. La technique offre une bonne extraction des composés phénoliques et des flavonoïdes utilisant l’eau comme solvant naturel. La quatrième et la dernière partie de ce travail s’est orientée vers l’optimisation et le développement d’une technologie « verte» utilisant les ultrasons et l’huile de tournesol comme solvant naturel pour l’extraction des composés aromatiques du thym. Cette nouvelle approche écologique permet l’extraction des absolues dépourvues de cire et des résidus de solvants pétroliers, contenant la teneur la plus élevée en thymol et exerçant la plus forte activité antioxydante
Plant essential oils and aromas are a real potential for the industry to substitute the synthetic compounds that might have harmful effects on the human health and the environment. In order to contribute to the green chemistry principles, this study focuses on the “eco-extraction” and valorization of natural plant extracts and the development of a new «green» technology for the extraction of aromatic compounds. The first part of this manuscript highlights the biological properties of the essential oils and aromas and the importance of their applications in various sectors especially in the food industry. In the second part, studies on the biological properties of the essential oils from plants of the Alliaceae family show their good antioxidant and antimicrobial activities. These encouraging results have allowed in the third part to evaluate the onion by-products resulting from the turbo hydrodistillation, which are normally considered as waste. The technique offers a good extraction of flavonoids and phenols using water as natural solvent. The fourth and final part of this work deals with the optimization and the development of a «green» technology using ultrasound and sunflower oil as a natural solvent for the extraction of aromatic compounds from thyme. This new ecological approach allows the extraction of absolutes free from waxes and petroleum solvent residues, having the highest content in thymol and exerting the strongest antioxidant activity

Ce travail constitue un recueil des mes activités d'enseignement et de recherche postdoctorales réalisées au sein du laboratoire " Chimie des Produits Naturels " (CPN) dans le cadre du projet de recherche " Ressources Naturels " soutenu par l'UMR CNRS 6134 " Sciences de l'Environnement " de l'Université de Corse Pasquale Paoli. Les travaux concernent la caractérisation des Plantes aromatiques et médicinales (PAM) et des produits de l'agroalimentaire au travers des mélanges complexes volatils qui en sont issus. Ces mélanges complexes sont des huiles essentielles, des hydrolats, des extraits aux solvants et des fractions volatiles. Les différentes étapes de la séquence analytique ont été examinées à savoir, le choix des végétaux et leur échantillonnage, la préparation de l'échantillon, son analyse proprement dite, l'interprétation des résultats au moyen de l'outil statistique et leur valorisation au travers de la recherche de principes actifs. La première partie vise à caractériser les huiles essentielles, les hydrolats, les extraits aux solvants et les fractions volatiles issus de PAM et ainsi que les arômes des huiles d'olives et de jus d'agrumes. Pour cela, nous avons examiné les potentialités de méthodes dites alternatives à l'hydrodistillation, méthode conventionnelle pour l'obtention des huiles essentielles et des hydrolats, telles que l'extraction assistée par micro-ondes et la MicroExtraction en Phase Solide. L'analyse proprement est réalisée au laboratoire CPN à l'aide de techniques chromatographiques telles que la Chromatographie en Phase Gazeuse (CPG) pour la quantification et la CPG couplée à la Spectrométrie de Masse (CPG-SM) pour l'identification des constituants des mélanges. Nous avons optimisé la séquence analytique en utilisant la complémentarité des techniques telles que la chromatographie sur colonne, l'utilisation de la SM en mode ionisation chimique et de la Résonance Magnétique Nucléaire pour l'identification de molécules absentes des bibliothèques de références. La deuxième partie concerne le développement d'un axe de recherche nouveau qui vise à mettre en évidence des principes actifs au travers de l'étude des propriétés antibactériennes et antifongiques des huiles essentielles et des extraits de végétaux. Nous avons mis en évidence les propriétés biologiques d'huiles essentielles et d'extraits sur un certain nombre de bactéries impliquées dans des infections nosocomiales et alimentaires. La dernière partie développe les perspectives de travail qui visent à renforcer les travaux sur les PAM et sur les produits identitaires de l'agroalimentaire produits en Corse en explorant de nouvelles matrices d'études (fraction lourde) et de nouvelles techniques d'extraction. La valorisation des mélanges complexes issus des végétaux par la recherche de nouveaux antibiotiques surpassant les mécanismes de résistance des bactéries et de nouveaux antioxydants reste un challenge scientifique d'avenir.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
CHAPTER 01: CHEMICAL STUDY OF VOLATILE ORGANIC COMPOUNDS Annona coriacea Mart. AND Xylopia aromatica (Lam.) Mart (Annonaceae) - In this research was performed the study of the chemical profile of the volatile organic compounds (VOCs) of flowers and leaves of species Annona coriacea and Xylopia aromatica (Annonaceae) by means of extraction in vivo and in vitro, through the techniques headspace-solid phase microextraction (HS-SPME), hydrodistillation and characterization by gas chromatography-mass spectrometry (GC-MS). By the analysis of the volatile fraction of flowers and leaves of A. coriacea it was observed different VOCs, according to the coatings utilized, since the selection of the fiber becomes one of the main factors for VOCs analysis in different vegetables parts. The chemical profile of the flowers of X. aromatica was obtained through HS-SPME, which compared the efficiency of in vivo and in vitro extractions, due to the higher number of VOCs present in the floral aroma. Once were obtained various chemical data, the multivariate analysis of the data was performed, which provided relevant information, contributing to the optimization of some conditions, such as selection of fiber, extraction time and extraction temperature yielded the characterization of the most number of VOCs. In this context, the conditions that presented the most promising results were: fibers CAR/PDMS and PA, extraction time equal the 60 min and extraction temperature of 29 °C. The essential oil of the flowers of X. aromatica was obtained by hydrodistillation and chemically characterized by GC-MS. The major compound present was the pentadecan-2-one (16.38%). The extraction techniques utilized were helpful, however there was qualitative and quantitative variation of VOCs, which is expected due to the extraction process be different. Thus, it is observed that depending on the objective of the survey or that searches for in the volatile fraction, it is necessary to use the technique to give the most promising results for the investigation held or in order to complement each other. In this way, the study of A. coriacea and X. aromatica contributed to the knowledge of the chemical profile of VOCs present in these species, seen from these investigations that the same can be applied in several industrial areas, for example cosmetic, food, among others.
CAPÍTULO 01: ESTUDO QUÍMICO DOS COMPOSTOS ORGÂNICOS VOLÁTEIS DE Annona coriacea Mart. E Xylopia aromatica (Lam.) Mart (Annonaceae) - Nesta pesquisa realizou-se o estudo do perfil químico dos compostos orgânicos voláteis (COVs) das flores e folhas das espécies Annona coriacea e Xylopia aromatica (Annonaceae), por meio da extração in vivo e in vitro, através das técnicas de headspace-microextração em fase sólida (HS-SPME), hidrodestilação e caracterização via cromatografia gasosa acoplada à espectrometria de massas (CG-EM). Pela análise da fração volátil das flores e folhas de A. coriacea observou-se COVs diferentes, de acordo com os revestimentos usados, visto que a seleção da fibra se torna um dos principais fatores para a análise de COVs em partes vegetais diferentes. O perfil químico das flores de X. aromatica foi obtido via HS-SPME, em que se comparou a eficiência das extrações in vivo e in vitro, em função do maior número de COVs presentes no aroma floral. Uma vez que foram obtidos vários dados químicos, foi realizada a análise multivariada dos mesmos, a qual forneceu informações relevantes, contribuindo para a otimização de algumas condições, tais como seleção da fibra, tempo de extração e temperatura de extração que rendessem a caracterização do maior número de COVs. Neste contexto, as condições que apresentaram os resultados mais promissores foram: fibras de CAR/PDMS e PA, tempo de extração igual a 60 min e temperatura de extração de 29 oC. O óleo essencial das flores de X. aromatica foi obtido por hidrodestilação e caracterizado quimicamente por CG-EM. O composto majoritário presente foi a pentadecan-2-ona (16,38%). As técnicas de extração utilizadas foram úteis, entretanto houve variação quali e quantitativa dos COVs, o que é esperado devido ao processo de extração ser diferente. Assim, observou-se que dependendo do objetivo da pesquisa ou do que se procura na fração volátil, faz-se necessária a utilização da técnica que dê os resultados mais promissores para a investigação realizada ou de forma que se complementem. Desta maneira, o estudo de A. coriacea e X. aromatica contribuiu para o conhecimento do perfil químico dos COVs presentes nestas espécies, visto que a partir destas investigações os mesmos podem ser aplicados em várias áreas industriais, por exemplo cosmético, alimentos, dentre outros.

Les régions de Midi-Pyrénées (France) et de Chongqing (Chine) sont riches en plantes aromatiques et médicinales dites oubliées (ou médiévales). Afin de valoriser pleinement les différentes bio-molécules extractibles de ces plantes, le concept de MAP-raffinerie a été créé et appliqué à une sélection de plantes issues de ces deux régions. Plusieurs technologies d’extraction utilisant l’eau comme solvant vert (hydrodistillation, distillation à la vapeur et extraction par eau sub-critique) ont ainsi été employées et leur impact tant sur la composition des huiles essentielles que sur la récupération des molécules anti-oxydantes a été évalué. Dans un premier temps, une liste de plantes aromatiques et médicinales oubliées, voire sous-utilisées dans les deux régions a été établie selon des règles de sélection prédéfinies. Six plantes modèles de la région de Midi-Pyrénées (Tussilago farfara L., Calendula arvensis L., Robinia pseudoacacia L., Geranium robertianum L., Cytisus scoparius L. et Spartium junceum L.) et trois plantes de la région de Chongqing (Tussilago farfara L., Citrus aurantium L. et Saussurea costus) ont finalement été retenues. Puis, le concept de MAP-raffinerie a été appliqué à ces plantes afin d’étudier leur possible valorisation globale. L’étude des compositions chimiques des extraits volatils des racines de Tussilago farfara L. et de Calendula arvensis L., ainsi que des boutons de fleurs de Spartium junceum L. a été réalisée par GC et GC-MS pour la première fois. Les principaux composés chimiques dans l’extrait volatil de racines de Tussilago farfara L. étaient des hydrocarbures sesquiterpéniques et des composés aliphatiques tandis que les principaux composés chimiques dans l’extrait volatil de racines de Calendula arvensis L. étaient des sesquiterpènes oxygénés, des monoterpènes oxygénés et des diterpènes oxygénés. L’extrait volatil de boutons de fleurs de Spartium junceum L. était principalement composé de composés aliphatiques. Par ailleurs, les résultats de l’évaluation des capacités anti-oxydantes des extraits (par les tests DPPH, ABTS, FRAP, ORAC et Folin-Ciocalteu) ont montrés que plusieurs plantes comme Cytisus scoparius L., Tussilago farfara L., Citrus aurantium L. ou Robinia pseudoacacia L. pourraient être des sources potentielles d’anti-oxydants naturels. D’un point de vue technologique, les comparaisons de l’utilisation de l’hydrodistillation (HD), de la distillation à la vapeur (SD) et de l’extraction par eau sub-critique (SWE) ont montrées que si la HD et la SD ont des effets limités sur la composition des huiles essentielles, la HD semble être une méthode plus efficace pour la récupération des composés anti-oxydants à partir des résidus de distillation que la SD tandis que la SWE s’avère être une technologie prometteuse pour l’extraction directe de ces molécules à partir des plantes. Si la composition minérale de l’eau lors de l’hydrodistillation n’a que des effets très limités sur les rendements d’extraction, les teneurs en ions calcium et bicarbonate des eaux ont par contre des effets décroissants significatifs sur la capacité anti-oxydante et sur la teneur phénolique totale des extraits aqueux et méthanoliques. Au vue de ces résultats, un concept amélioré de MAP-raffinerie a été développé en intégrant une extraction à l’eau sub-critique pour l’extraction des composés anti-oxydants des résidus d’extraction primaire. Selon ce nouveau concept, cinq extraits peuvent être obtenus à partir des matières végétales: un extrait volatil, un extrait aqueux, un extrait méthanolique, un extrait à l’eau sub-critique et in fine un résidu solide. Les premiers résultats ont montrés que la "MAP-raffinerie améliorée" augmente de manière significative la récupération des antioxydants par rapport à la MAP-raffinerie originale et permet d’envisager une valorisation plus facile du résidu solide en agro-matériaux du fait de sa faible teneur en eau résiduelle
In both Midi-Pyrénées region (France) and Chongqing region (China), there are rich and underutilized medicinal and aromatic plants (MAP). Aiming at fully exploiting different molecules in these plants, the concept of MAP-refinery was developed and applied to several underutilized medicinal and aromatic plants in these two regions. Several water-based green extraction technologies of natural products (e.g. hydrodistillation, steam distillation and subcritical water extraction) were also investigated to look at their effects on essential oil composition and antioxidants recovery from selected plants. Firstly, lists of forgotten or underutilized medicinal and aromatic plants in both regions were established according to the rules of selection. From the lists, six plants in the Midi-Pyrénées region (Tussilago farfara L., Calendula arvensis L., Robinia pseudoacacia L., Geranium robertianum L., Cytisus scoparius L. and Spartium junceum L.) and three plants in the Chongqing region (Tussilago farfara L., Citrum aurantium L. and Saussurea costus) were finally selected for investigations. Then the MAP-refinery was applied to the selected plants in two regions in order to realise their global valorisation. Volatile extracts composition in the roots of Tussilago farfara L. and Calendula arvensis L., as well as flower buds of Spartium junceum L. were firstly investigated. The main chemical compounds in volatile extract from Tussilago farfara L. roots were sesquiterpene hydrocarbons and aliphatic compounds while main chemical compounds in volatile extract from Calendula arvensis L. roots were oxygenated sesquiterpenes, oxygenated monoterpenes and oxygenated diterpenes. The volatile extract from flower buds of Spartium junceum L. was mainly composed of aliphatic compounds. Antioxidant capacity evaluation results (by DPPH, ABTS, FRAC, ORAC and Folin-Ciocalteu tests) showed that several plant samples like Cytisus scoparius L., Tussilago farfara L., Citrum aurantium L. and Robinia pseudoacacia L. could be potential sources of natural antioxidants. Comparisons of hydrodistillation (HD), steam distillation (SD) and subcritical water extraction (SWE) showed that HD and SD had limited effects on essential oil composition but HD, SD and SWE had significant impacts on the recovery of antioxidants. Hydrodistillation seemed to be a better method for recovery of antioxidant compounds from residues of distillation than steam distillation. However, SWE appeared to be a more efficient method for direct extraction of antioxidant molecules (or phenolic compounds) from plants. In the hydrodistillation process, mineral contents in water were found to have very limited effects on yields of extracts but calcium and bicarbonate ions, had significant decreasing effects on antioxidant capacity and total phenolic content of both aqueous and methanolic extracts. Finally, an improved MAP-refinery was developed. Subcritical water was used for further extraction of antioxidant compounds from residues in original MAP-refinery. In this way, five parts could be obtained from plant materials: volatile extract, aqueous extract, methanolic extract, subcritical water extract and the final residue. The results showed that the improved MAP-refinery significantly increased the recovery of antioxidants compared with original MAP-refinery. This promising process will also allow a better valorisation of the final solid residue due to the lower content of residual water

碩士
國立成功大學
化學工程學系
107
In this study, the response surface methodology (RSM) was adopted to evaluate the effects of process parameters of hydrodistillation for extraction of tea tree oil (TTO) in presence of a nonionic surfactant - Tween 80. These process parameters include the surfactant concentration, the extraction time and the liquid/solid ratio on the extraction yield of TTO were investigated. Results showed that the data were adequately fitted into the second-order polynomial model, in which the extraction time had a significant effect on the extraction yield of TTO. It was predicted that the optimum extraction condition is 2700 ppm as the concentration of Tween 80 in hydro-distill, 140 min for extraction time and liquid/solid ratio at 11.6. Under these optimal conditions, the extraction yield of tea tee oil was 5.48%. The kinetics model of oil extraction comprised two stages of washing and diffusion. The surfactant added could increase both kinetic parameters. The components of tea tree oil were analyzed with gas chromatography (GC), while HPLC was employed to analyze the quantity of Tween 80 present in the extracted TTO. The oil obtained under the optimal extraction condition underwent further tests for the stability of its microemulsion formulation with Tween 80, as well as its antibacterial and antioxidant properties. The maximum amount of tea tree oil that could be solubilized within the micelles increased with an increasing concentration of Tween 80. The microemulsion made with TTO in 5 wt% Tween 80 was stable for over 30 days. Antioxidant activity of tea tree oil was determined using the DPPH assay. The EC50 value was 75.07 mg/g. The -terpinene, one main ingredient of TTO, exhibited good antioxidant activity, while its EC50 was 45.85 mg/g. The antibacterial activity of tea tree oil was evaluated against Escherichia coli and Staphylococcus aureus with the agar disk-diffusion method and the broth dilution. Tea tree oil displayed the superior antimicrobial properties evaluated by the disk-diffusion assay. The MBC values for tea tree oil against E. coli and S. aureus were 8.77 and 6.08 mg/mL, respectively.

碩士
弘光科技大學
化妝品科技研究所
104
Tagetes lemmoniia, an aromatic plant, the whole plant has a special flavor and has been used in the food, cosmetics, and other multi-purpose. In this study, solvent-free microwave extraction (SFME) and hydro-distillation extraction (HDE) were used to extract essential oils form leaves and flower of Tagetes lemmoniia and the chemical composition were analyzed by gas chromatography and mass spectrometry (GC- MS).The essential oils recovered by SFME (30 min, 500W) and HDE (2h) were 0.6% and 0.5%, respectively. The main components of essential oils for SFME were 3-pentadecyl ester (29.59%), verbenone (23.5%), limonene (13.13%), linalool (11.47%) and trans-tagetone (10.79%). The main component of essential oils for HDE was 3-pentadecyl ester up to 62.61%, while the content of verbenone was only 1.77%. Antioxidant capacity of essential oils extracted by SFME was better than that of HDE. The total phenolic content of SFME essential oils was 138.17mg GAE/g sample which was also higher than the HDE oils of 63.04 mg GAE/g sample. Overall, leaves essential oil of Tagetes lemmoniia obtained by SFME will have higher extraction yield, a shorter extraction time, and also exhibited a strong antioxidant capacity. The high antioxidant capacity of essential oil obtained SFME may be related to a high proportion of verbenone. SFME technology on essential oil extraction will enable the application of Tagetes lemmoniia on cosmetics not only more environmentally-friendly but more efficacy.

Mentha longifolia subsp. polyadena was collected from seven localities in South Africa and from a single population in Botswana to study the essential oil composition and antimicrobial activity of this ethnomedicinal plant. The essential oils were obtained by hydrodistillation and analysed by gas chromatography coupled to mass spectroscopy (GC/MS) and a cluster analysis was performed on the essential oil dataset. From eight samples (representing eight natural populations), two major chemotypes were identified: (i) a menthofuran rich type (51.4% - 61.6%); and (ii) a cis-piperitone epoxide (14.7% – 35.7%) and piperitenone oxide (14.6% - 65.7%) rich type. The constituent analysis showed quantitative variation with higher amounts of oxygencontaining monoterpenes ranging from 56.5% to 89.6% whilst the sesquiterpene hydrocarbons ranged from 4.4% to 16.7%. The essential oil from the different localities mostly showed moderate to good antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Bacillus cereus, Moraxella catarrhalis, Yersinia enterocolitica and Enterococcus faecalis. The oils were generally inactive against Escherichia coli and Salmonella typhimurium. Candida albicans and Cryptococcus neoformans indicated highest sensitivities for oil samples from Komukwane (3 mg/ml and 0.5 mg/ml respectively) and Prins Albert (0.5 mg/ml and 1.6 mg/ml respectively). The HPLC profiles of the methanol and chloroform (1:1) extracts were more conservative and less variable compared to the essential oils. Two major peaks corresponding to retention times of 22.39 min and 26.47 min were present in all eight samples. Most of the solvent extracts displayed moderate to good antimicrobial activity against Gram-positive pathogens, in particular against S. aureus, S. epidermidis and B. cereus with MIC values ranging from 0.5 mg/ml to 2 mg/ml in most cases. The extracts also demonstrated moderate to good activity against most of the Gram-negative pathogens, in particular against Y. eneterocolitica and M. catarrhalis, with MIC values ranging from 0.5 mg/ml to 2 mg/ml. These results may in part provide scientific evidence for the extensive use of Mentha longifolia in traditional healing.

Dissertação de Mestrado em Segurança Alimentar apresentada à Faculdade de Farmácia
Crithmum maritimum é uma planta da família Apiaceae, típica da zona do Mediterrâneo, usada como alimento fresco, na medicina tradicional e com aplicações mais recentes na indústria cosmética. As suas potenciais utilizações não estão totalmente descritas cientificamente, pelo que se justifica ampliar o seu estudo. Contudo, há já algumas atividades biológicas associadas ao seu óleo essencial. Neste contexto, o objetivo deste trabalho consistiu em valorizar o óleo essencial (OE) bem como a água residual da hidrodestilação (AHD), rica em compostos fenólicos, não só pela sua caraterização química como também pela avaliação das suas atividades antioxidante, anti-inflamatória e anti-fúngica. O OE foi obtido por hidrodestilação e a AHD resultante deste processo extrativo foi sujeita a evaporação a vácuo e liofilização, tendo sido posteriormente caraterizada com recurso a cromatografia em camada fina (TLC) e cromatografia de alta resolução com deteção de fotodíodos (HPLC-DAD). A AHD evidenciou um perfil rico em ácidos fenólicos do tipo hidroxicinâmico e pequenas quantidades de flavonóides. A caraterização do OE foi obtida por GC e GC-MS e permitiu identificar como constituintes maioritários γ-terpineno, sabineno e timol metil éter.Foram avaliadas as propriedades antifúngicas do OE e da AHD no que diz respeito a leveduras e fungos filamentosos (Candida spp., Cryptococcus neoformans, dermatófitos e Aspergillus spp.), conhecidos como agentes patogénicos de patologias fúngicas prevalentes. A avaliação da concentração mínima inibitória (MIC) e da concentração letal mínima (MLC) permitiu concluir que o OE de C. maritimum tem uma atividade significativa, especialmente em Cryptococcus neoformans (com MIC de 0.26 mg/mL e MLC de 0.52 mg/mL), Epidermophyton floccosum e Microsporum canis (MIC e MLC com valor de 0.52 mg/mL). Adicionalmente o óleo demonstrou inibir o tubo germinativo e a formação de biofilmes de C. albicans, sugerindo um potencial interesse do óleo essencial de C. maritimum para o tratamento de candidíase, associado à presença dos seus compostos maioritários. A AHD demonstrou não exercer atividade antifúngica para as estirpes testadas (MIC e MLC >5 mg/mL).Tendo em conta que dados de estudos prévios sublinhavam a atividade antioxidante diminuta do OE e um potencial antioxidante de relevância para extratos de compostos não voláteis, associados à presença de ácido clorogénico, fez-se um estudo de atividade antioxidante baseado na capacidade de neutralização do radical DPPH apenas para a AHD, tendo o extrato apresentado um valor de IC50 de 0.68 ± 0.16 mg/mL. O estudo realizado demonstrou também que esta atividade é essencialmente devida à presença de ácidos fenólicos, principalmente ao ácido clorogénico mas também ao flavonol presente no extrato.A atividade anti-inflamatória, relativamente à qual não havia qualquer estudo nem em OE nem em extratos fenólicos de C. maritimum, foi avaliada. Os resultados permitiram concluir que o OE apresentou um IC50 de 0.01225 ± 0.00192 mg/mL. O óleo essencial reduziu significativamente a produção de óxido nítrico em macrófagos estimulados com LPS a uma concentração de 0.003 mg/mL sem comprometer a viabilidade celular. Esta atividade do OE poderá ser atribuída à atividade dos compostos maioritários, já que estes demonstraram também ter atividade. Relativamente à AHD, esta não demonstrou ter atividade anti-inflamatória. Em conclusão, este trabalho permitiu avaliar pela primeira vez o potencial anti-inflamatório do óleo essencial de C. maritimum, bem como a sua atividade antifúngica no que diz respeito aos fatores de virulência de C. albicans. Permitiu ainda evidenciar, pela primeira vez, a composição fenólica da AHD, bem como o seu potencial biológico no que diz respeito à sua atividade antioxidante. Estes resultados salientam a importância do reaproveitamento da água resultante do processo de extração do óleo essencial de C. maritimum, como fonte de compostos bioativos, atribuindo assim um valor adicional a esta planta aromática. Todo o estudo evidencia a possibilidade de uma maior utilização de Crithmum maritimum, com aplicações alimentares e medicinais.
Crithmum maritimum is a plant of the Apiaceae family, typical of the Mediterranean zone, used as fresh food, in traditional medicine and with more recent applications in the cosmetic industry. Its potential uses are not fully described scientifically, and it is therefore justified to extend its study. However, there are already some biological activities associated with its essential oil. In this context, the objective of this work was to evaluate the essential oil (EO) as well as the hydrodistillation residual water (HRW), rich in phenolic compounds, not only for its chemical characterization but also for the evaluation of its antioxidant, anti-inflammatory and antifungal activities. The EO was obtained by hydrodistillation and the HRW resulting from this extraction process was subjected to vacuum evaporation and freeze-drying and was later characterized by thin layer chromatography (TLC) and high resolution chromatography with photodiode detection (HPLC-DAD). HRW showed a profile rich in hydroxycinnamic phenolic acids and small amounts of flavonoids. The EO characterization was obtained by GC and GC-MS and allowed the identification of γ-terpinene, sabinene and thymol methyl ether as major constituents. The antifungal properties of the EO and HRW were evaluated for yeast and filamentous fungi (Candida spp., Cryptococcus neoformans, dermatophytes and Aspergillus spp.), known as pathogens of prevalent fungal pathologies. The evaluation of minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) allowed us to conclude that C. maritimum EO has a significant activity, especially in Cryptococcus neoformans (with MIC of 0.26 mg / mL and MLC of 0.52 mg/mL), Epidermophyton floccosum and Microsporum canis (MIC and MLC with a value of 0.52 mg/mL). In addition, the oil has proved to inhibit the germ tube and biofilm formation of C. albicans, suggesting a potential interest of C. maritimum essential oil for the treatment of candidiasis, associated with the presence of its major compounds. AHD demonstrated no antifungal activity for the strains tested (MIC and MLC> 5 mg/mL). Considering that data from previous studies highlighted the low antioxidant activity of EO and a relevant antioxidant potential for extracts of non-volatile compounds associated with the presence of chlorogenic acid, a study of antioxidant activity was conducted based on the neutralizing capacity of the radical DPPH only for HRW. As a result, the extract presented an IC50 value of 0.68 ± 0.16 mg/mL. The study also demonstrated that this activity is essentially due to the presence of phenolic acids, mainly chlorogenic acid but also to the flavonol present in the extract. The anti-inflammatory activity, for which there was no study in either the EO or phenolic extracts of C. maritimum, was evaluated. The results allowed to conclude that EO presented an IC50 of 0.01225 ± 0.00192 mg/mL. The essential oil significantly reduced the production of nitric oxide in LPS-stimulated macrophages at a concentration of 0.003 mg/mL without compromising cell viability. This activity of EO can be attributed to the activity of the major compounds, since these have also shown to have activity. Regarding HRW, it has not shown any anti-inflammatory activity. In conclusion, this work allowed to evaluate for the first time the anti-inflammatory potential of the essential oil of C. maritimum, as well as its antifungal activity with respect to the virulence factors of C. albicans. It also allowed to demonstrate, for the first time, the phenolic composition of HRW, as well as its biological potential with respect to its antioxidant activity. These results highlight the importance of the reuse of water resulting from the extraction process of the essential oil of C. maritimum as a source of bioactive compounds, thus giving an additional value to this aromatic plant. The whole study evidences the possibility of a greater use of Crithmum maritimum, with food and healthcare applications.