This study involved the isolation, characterization, quantification and evaluation of the enantiomeric purity of linalool in the essential oil of aromatic species Lippia alba (Mill.) NE Brown (chemotype linalol-1,8-cineole), Ocimum basilicum L., Pelargonium graveolens L'Herit, Cinnamomum camphora Nees and Eberm var. linaloolifera Fujita, Aniba rosaeodora Ducke and Coriandrum sativum L., and carvone and limonene in the essential of Lippia alba (chemotype carvone-limonene). The quantification of monoterpenes was performed by GC-FID method for the internal standard. The analytical curves for quantification were linear in the range 1.0 to 10.0 mg mL-1 and showed appropriate values of correlation coefficients (0.996 to 0.999). The linalool was found with 91.38% (w/w) in essential oil of A. rosaeodora, 84.00% in C. camphora, 79.25% in L. alba, 79.00% in C. sativum, 73.33% in O. basilicum and 13.60% in P. graveolens. The carvone showed 58.13% and limonene showed 31.71% (w/w) of essential oil of L. alba. The enantiomeric purity analysis performed by GC-FID, using a column with enantioselective phase (β-cyclodextrin) and co-injection with commercial patterns, revealed the presence of the enantiomer (S)-(+)-linalool in L. alba and (R)-(-)-linalool in O. basilicum and C. camphora. The two enantiomeric forms of linalool were found in the P. graveolens, A. rosaeodora and C. sativum species. In the L. alba species (chemotype carvone-limonene) was observed a single enantiomeric form (R)-(-)-carvone and limonene showed the enantiomeric forms (S)-(-)-limonene and (R)-(+)-limonene, with predominance of the latter. The linalool was isolated from the essential oil of L. alba by TLC and subjected to analysis of GC-MS, GC-FID, 1H and 13C NMR and [α]D, which elucidated its structure and stereochemistry. The (S)-(+)-linalool isolated from L. alba (> 99%) was used to obtain the analytical curve. This isolation was necessary due to unavailability of acquire the (S)-(+)-linalool commercially pure. / Este estudo envolveu isolamento, caracterização, quantificação e avaliação da pureza enantiomérica do linalol nos óleos essenciais das espécies aromáticas Lippia alba (Mill.) N. E. Brown (quimiotipo linalol-1,8-cineol), Ocimum basilicum L, Pelargonium graveolens L Herit, Cinnamomum camphora Nees e Eberm var. linaloolifera Fujita, Aniba rosaeodora Ducke e Coriandrum sativum L. e da carvona e do limoneno no óleo essencial de Lippia alba (Mill.) N. E. Brown (quimiotipo carvona-limoneno). A quantificação dos monoterpenos foi realizada por CG-DIC pelo método do padrão interno. As curvas analíticas para a quantificação foram lineares no intervalo de 1,0 a 10,0 mg mL-1 e apresentaram valores adequados de coeficientes de correlação (0,996 a 0,999). O linalol foi encontrado com 91,38% (m/m) no óleo essencial de A. rosaeodora, 84,00% em C. camphora, 79,25% em L. alba, 79,00% em C. sativum, 73,33% em O. basilicum e 13,60% em P. graveolens. A carvona apresentou 58,13% e o limoneno 31,71% (m/m) no óleo essencial de L. alba (quimiotipo carvona-limoneno). As análises de purezas enantioméricas realizadas por CG-DIC, utilizando uma coluna com fase enantiosseletiva (β-ciclodextrina) e a co-injeção com padrões comerciais, revelaram a presença do enantiômero (S)-(+)-linalol em L. alba e do (R)-(-)-linalol em O. basilicum e em C. camphora. As duas formas enantioméricas do linalol foram encontradas nas espécies P. graveolens, A. rosaeodora e C. sativum. Na espécie L. alba (quimiotipo carvona-limoneno) foi observada uma única forma enantiomérica (R)-(-)-carvona e o limoneno apresentou as formas enantioméricas (S)-(-)-limoneno e (R)-(+)-limoneno com predominância desta última. O linalol foi isolado do óleo essencial de L. alba por CCD e submetido à análise por CG-EM, CG-DIC, RMN de 1H e 13C e [α]D, as quais elucidaram sua estrutura e estereoquímica. O (S)-(+)-linalol isolado de L. alba (> 99%) foi utilizado na obtenção da curva analítica. Esse isolamento foi necessário devido à indisponibilidade em adquirir o (S)-(+)-linalol comercialmente puro.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The sustainable development of northern agroecosystems requires, among other factors, the use of species adapted to region the edafoclimtic conditions. Plants able to support water deficits are more indicated, considering to no regular distribution of pluviometric precipitation the high evapotranspiration are characteristic northest region of Brazil. The present work was carried out to determine the ecofisiologic and biochemistry behavoir of three cultivars of basil, Ocimum basilicum L. (Maria Bonita, Mara and Genovese) cultivated under water stress. The ecofisiologics variables were: hidric potential of leaves, photossintesis, transpiration, stomatal conductance; the biochemitry variables were: soluble carbohydrates and proteins, totals cloroplasts, tenor of essential oil and percentage of linalool. The varibles were analyzed in plants with four months of aged, cultivated in 24 vases of 17L, in greenhouse, having two treatments: T0 (control) and T1 (experimental group), with four repetitions. The variables were evaluated at 08:30 to 10:30 hours every two days, during 15 days in October 2005. In base of results it was identifyed that basil presents mechanisms of adaptation under water stress minimizing the water lose. Mara’and Maria Bonita’cultivars presented more adaptaded under water stress in comparison with ‘Genovese’cultivar. In relation to the essential linalool content the three cultivars kept their yield even under water stress. It was observed the geraniol presence in Maria Bonita cultivar. / O desenvolvimento sustentável dos agroecossistemas nordestinos requer, dentre outros fatores, o uso de espécies adaptadas às condições edafomorfoclimáticas da região. Plantas capazes de suportar déficits hídricos são mais indicadas, uma vez que a distribuição irregular da precipitação pluviométrica, aliada a alta evapotranspiração são características do Nordeste brasileiro. O presente trabalho objetiva determinar o comportamento ecofisiológico e análise bioquímica de três cultivares de manjericão, Ocimum basilicum L.(‘Maria Bonita’, ‘Mara’ e ‘Genovese’) quando cultivadas sob estresse hídrico. As variáveis ecofisiológicas foram: potencial hídrico foliar, fotossíntese, transpiração, condutância estomática; e as bioquímicas foram: carboidratos solúveis, proteínas solúveis, cloroplastos totais e teor de óleo essencial e de linalol no óleo essencial. As variáveis ecofisiológicas e bioquímicas foram analisadas quando as plantas apresentavam 4 meses de idade. Cultivadas em 24 vasos de 17 L, em estufa agrícola, sendo 2 tratamentos: T0 (testemunha) e T1 (grupo experimental), com 4 repetições. As variáveis ecofisiológicas foram avaliadas entre 08:30 a 10:30 horas, em dias alternados, durante 15 do mês de outubro de 2005. Com base nos resultados obtidos, constatou-se que o manjericão apresenta mecanismos de adaptação, que sob condição de estresse hídrico, diminuem as perdas de água. As cultivares ‘Mara’ e ‘Maria Bonita’ apresentaram adaptadas às condições de estresse hídrico quando comparadas com a cultivar ‘Genovese’. Em relação ao teor linalol as três cultivares mantiveram as suas produtividades mesmo sob condição de estresse. Observou-se também, a presença de geraniol no óleo essencial da cultivar Maria Bonita.

Tesis por compendio / [EN] In the present Doctoral Thesis, antimicrobial active packaging materials, at lab and at semi-industrial scale, have been developed with the aim to reduce the natural flora of peeled and cut fruit and extend its shelf life. Packaging prototypes have been developed for their further application. Prior to developing the active materials, the most suitable active agents were selected. To that end, the antimicrobial properties of the volatile active agents citral, hexanal and linalool and mixtures thereof were evaluated against typical microorganisms related to fruit spoilage, molds and yeast, concluding that the effectiveness of the mixture is higher than the sum of the effectiveness of the individual agents. Likewise, non-volatile antimicrobial agents such as potassium sorbate and sodium benzoate were selected, which are widely used in the food industry due to their antifungal properties. With the selected active agents, monolayer polypropylene (PP) films with different concentration of the active mixture citral, hexanal and linalool, at lab scale by means of extrusion, and bilayer films at semi-industrial scale with different active layer thickness by means of coextrusion were prepared. Besides, active packaging trays were developed at semi-industrial scale by thermoforming active sheets obtained by coextrusion of PP and ethyl vinyl acetate (EVA) compounds containing potassium sorbate and sodium benzoate as active agents. Mechanical, barrier and thermal properties of the developed active packaging materials, as well as their sealability and transparency were evaluated. In general, the materials' properties were not affected in a significant manner. However, active trays decreased in transparency due to the incorporation of non-volatile active agents. The release kinetics of the volatile and non-volatile active agents were studied at different temperatures, defining their diffusion coefficients by the adjustment to mathematic models based on Second's Law Fick. Among the volatile active agents, hexanal showed a higher diffusion coefficient, followed by citral and linalool. On the other hand, very small differences were observed between potassium sorbate and sodium benzoate diffusion coefficients, being of the same order of magnitude. In vitro tests were also performed at different temperatures to evaluate the antimicrobial properties of the developed materials. In general, the active packaging materials showed high antimicrobial properties which were enhanced with the increment of temperature. Once the properties of the developed materials were evaluated, in vivo tests with peeled and cut orange and pineapple were performed by packing these fruits with the active film, active tray and their combination (active packaging system). In general, the active packaging system improved the microbiological preservation of the fruit for longer times, between 2 and 7 days for orange and pineapple, respectively, and maintained quality parameters of the fruit at stable levels for longer times. Lastly, the safety of the active packaging materials was evaluated according to the European food contact materials and food legislation, and it was concluded that these materials were not of any safety concern for the consumers. / [ES] En la presente Tesis Doctoral se han desarrollado materiales de envase activo antimicrobiano, a escala laboratorio y a escala semi-industrial, con el objetivo de reducir la proliferación de la flora natural de la fruta pelada y cortada y extender su vida útil. Se han desarrollo distintos prototipos para su posterior aplicación industrial Previo al desarrollo de los materiales de envase, se ha realizado una selección de agentes activos más idóneos. Para ello se han estudiado mediante ensayos in vitro las propiedades antimicrobianas de agentes activos volátiles, citral, hexanal y linalool y diferentes mezclas de los mismos, frente a distintos microorganismos típicos del deterioro de las frutas, mohos y levaduras, concluyendo que la efectividad de la mezcla de los tres es superior a la suma de la efectividad de los activos de forma individual. Así mismo, también se han seleccionado antimicrobianos no volátiles como el sorbato potásico y benzoato sódico, los cuáles son ampliamente empleados en la industria alimentaria debido principalmente a sus propiedades antifúngicas. Con los agentes activos seleccionados, se han desarrollado películas monocapa de polipropileno (PP) con distintas concentraciones de la mezcla activa, citral, hexanal y linalool, a escala laboratorio, mediante técnicas de extrusión, y películas bicapa a escala semi-industrial con distintos espesores de capa activa mediante coextrusión. Por otra parte, se desarrollaron bandejas activas a escala semi-industrial mediante termoconformado de láminas obtenidas por coextrusión de compuestos de PP y etilvinilaceteto (EVA) con sorbato potásico o benzoato sódico como agentes antimicrobianos. Se han evaluado las propiedades mecánicas, barrera y térmicas de los materiales activos desarrollados, así como su sellabilidad y transparencia. En general, las propiedades de los polímeros no se vieron afectadas de manera relevante. Sin embargo, las bandejas activas perdieron su carácter transparente debido a la incorporación de los agentes activos no volátiles. Se ha estudiado la cinética de liberación de los compuestos activos volátiles y no volátiles a distintas temperaturas, determinando los coeficientes de difusión de los agentes activos mediante el ajuste a modelos matemáticos de difusión basados en la Segunda Ley de Fick. Entre los agentes volátiles, el hexanal mostró un mayor coeficiente de difusión seguido de citral y linalool. Por otra parte, no hubo apenas diferencia en los coeficientes de difusión del sorbato potásico y benzoato sódico, siendo éstos del mismo orden de magnitud. Igualmente, se han realizado diferentes experimentos in vitro a distintas temperaturas para determinar las propiedades antimicrobianas de los materiales desarrollados. En general, los materiales activos presentan una elevada capacidad antimicrobiana que se ve potenciada al aumentar la temperatura de exposición. Una vez evaluadas las características de los materiales desarrollados, se han efectuado ensayos de envasado de naranja y piña pelada y cortada con las películas y las bandejas activas y con la combinación del sistema de envase bandeja activa termosellada con la película activa. En general, el sistema de envase activo mejoró la conservación de la fruta por un mayor tiempo, entre 2 y 7 días para la naranja y piña, respectivamente, presentando una gran capacidad antimicrobiana y manteniendo los parámetros de calidad de la fruta en niveles estables por un mayor tiempo. Por último, se ha estudiado la seguridad de estos materiales de acuerdo a la legislación de materiales en contacto con alimentos y la legislación alimentaria europea, concluyendo que los materiales activos desarrollados no presentan preocupación para la seguridad de los consumidores. / [CA] En la present Tesi Doctoral s'han desenvolupat materials d'envasament actiu antimicrobià, a escala de laboratori i a escala semi-industrial amb l'objectiu de reduir la proliferació de la flora natural de la fruita pelada i tallada i estendre la seua vida útil. S'han desenvolupament diferents prototips per a la seua posterior aplicació industrial. Previ al desenvolupament dels materials actius, s'han seleccionat els agents actius mes idonis estudiant mitjançant assajos in vitro les propietats antimicrobianes d'agents actius volàtils, citral, hexanal i linalool i diferents mescles dels mateixos, enfront de diferents microorganismes típics de la deterioració de les fruites -floridures i llevats- concloent que l'efectivitat de la mescla dels tres és superior a la suma de l'efectivitat dels actius de forma individual. Així mateix, s'han seleccionat antimicrobians no volàtils, sorbat potàssic i benzoat sòdic, els quals son àmpliament empleats a l'industria alimentaria per les seues propietats antifúngiques. Amb els agents actius seleccionats, s'han desenvolupat pel·lícules monocapa de polipropilè (PP) amb diferents concentracions de la mescla activa, citral, hexanal i linalool, a escala laboratori, mitjançant tècniques d'extrusió, i pel·lícules bicapa a escala semi-industrial amb diferents espessors de capa activa mitjançant coextrusió. D'altra banda, s'han desenvolupat safates actives a escala semi-industrial mitjançant termoconformació de làmines obtingudes per coextrusió de compostos de PP i etil vinil acetat (EVA) amb sorbat potàssic o benzoat sòdic com a agents antimicrobians. S'han avaluat les propietats mecàniques, barrera i tèrmiques dels materials actius desenvolupats, així com la seua sellabilidad i transparència. En general, les propietats dels polímers no es van veure afectades de manera rellevant. No obstant això, les safates actives van perdre el seu caràcter transparent a causa de la incorporació dels agents actius no volàtils. S'ha estudiat la cinètica d'alliberament dels compostos actius volàtils i no volàtils a diferents temperatures, determinant els coeficients de difusió dels agents actius mitjançant l'ajust a models matemàtics de difusió basats en la Segona Llei de Fick. Entre els agents volàtils, l' hexanal va mostrar un major coeficient de difusió seguit de citral i linalool. D'altra banda, no va haver-hi a penes diferències en els coeficients de difusió del sorbat potàssic i benzoat sòdic, sent aquests del mateix ordre de magnitud. Igualment, s'han realitzat diferents experiments in vitro a diferents temperatures per determinar les propietats antimicrobianes dels materials desenvolupats. En general, els materials actius presenten una elevada capacitat antimicrobiana que es veu potenciada en augmentar la temperatura d'exposició. Una vegada avaluades les característiques dels materials desenvolupats s'han efectuat assajos d'envasament de taronja i pinya pelada i tallada amb la safata, la pel·lícula activa i la seva combinació (sistema d'envàs actiu). En general, el sistema d'envàs actiu va millorar la conservació de la fruita per un major temps, entre 2 i 7 dies per a la taronja i pinya respectivament, presentant una gran capacitat antimicrobiana i mantenint els paràmetres de qualitat de la fruita en nivells estables per un major temps. Finalment, s'ha estudiat la seguretat d'aquests materials d'acord a la legislació de materials en contacte amb aliments i la legislació alimentària europea, concloent que els materials actius desenvolupats no presenten preocupació per a la seguretat dels consumidors. / Lara Lledó, MI. (2016). Antimicrobial packaging system for minimally processed fruit [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/61388 / TESIS / Premios Extraordinarios de tesis doctorales / Compendio

L’espace sensoriel des vins secs de Riesling a été établi en utilisant diverses méthodologies d’analyse sensorielle. Cela a été possible grâce à la comparaison avec des vins blancs secs de styles différents, en particulier avec des vins blancs secs de Bordeaux montrant une caractéristique aromatique forte du Sauvignon blanc. Deux panels d’experts, l’un en France, l’autre en Allemagne ont démontré que la typicité aromatique des vins Riesling est particulièrement associée à des descripteurs fruités comme les notes d’agrume et de fruits jaunes. Des corrélations ont aussi été établies entre les données sensorielles et l’analyse quantitative des composés volatils déterminés par chromatographie en phase gazeuse couplée à la spectrométrie de masse (CG-SM). En particulier, pour la première fois, il a été possible de démontrer que les teneurs en 3 sulfanylhexan 1-ol (3SH) sont significativement corrélées avec le jugement de typicité des vins de Riesling. Ensuite, l’impact de conditions viticoles (régime hydrique de la vigne, pratique de l’effeuillage) pendant la période végétative a été mesuré vis à vis des teneurs en précurseurs d’arômes des raisins et des moûts ainsi que pour des composants aromatiques des vins. Ainsi, les concentrations en composés volatils tels que le 3SH, le linalol et le 1,1,6-trimethyl-1,2-dihydronaphthalene (TDN) dans les vins sont évidemment influencées par des facteurs viticoles. Par ailleurs, dans une démarche de caractérisation de composés de l’arôme des vins de Riesling, un composé bien connu, le trans-cinnamate d’éthyle, a été identifié comme étant susceptible d’influencer l’arôme des vins et d’être impliqué dans la perception de leur typicité aromatique. La méthodologie appliquée pour arriver à ce résultat, a intégré à la fois le fractionnement par chromatographie en phase liquide de haute performance (CLHP) en associant une approche sensorielle comparative entre les extraits de vins Riesling et ceux de vins blancs secs de Bordeaux, puis la chromatographie en phase gazeuse en deux dimensions en mode de détection par olfactométrie et spectrométrie de masse de haute résolution (2D-CG-O-HRMS). Le couplage de ces deux approches a permis de constituer des outils suffisamment pertinents pour décrypter l’arôme du Riesling. L’ensemble de ces résultats a permis d’accroitre nos connaissances sur l’expression aromatique des vins de Riesling et les composés aromatiques impliqués ainsi que sur l’impact des conditions viticoles. / The sensory concept of typicality of dry Riesling wines was demonstrated using different sensory analysis methodologies. This was achieved by comparison to other styles of dry white wines especially from Bordeaux, which showed strong Sauvignon blanc character. ‘Fruity’ descriptors like ‘citrus fruit’ and ‘yellow fruit’ were shown to be related to dry Riesling wines’ typicality independently by two panels of wine experts. Analysis of volatiles by means of gas chromatography coupled to mass selective detection (GC-MSD) was realised in order to link sensory data to aroma compounds’ concentrations. For the first time, concentrations of the high potent aroma compound 3 sulfanylhexan-1-ol (3SH) were shown to correlate with typicality ratings for Riesling wines. The impact of viticultural conditions during the grape vine growing period on finished wines’ aroma was studied with respect to grape vine water status and leaf removal practices including precursor studies in berries and must. It was clearly shown that changing viticultural conditions influenced volatile aroma compounds’ concentrations in finished wines, especially for 3SH, linalool and 1,1,6 trimethyl-1,2-dihydronaphthalene (TDN). Additionally the well-known aroma compound trans-ethyl cinnamate was identified to be a possible impacting aroma compound implicated in Riesling wines’ aromatic typicality. The applied methodology using two-dimensional gas chromatography with olfactometric and high resolution mass spectrometric detection (2D-GC-O/HRMS) after a sensory guided fractionation using high pressure liquid chromatography (HPLC) in a comparative approach to dry white wines from Bordeaux, showed to be a powerful tool to decrypt Riesling wines’ aroma. The totality of results allowed improving the knowledge on aromatic typicality of Riesling wines and the imparted aroma compounds and how those concentrations could be influenced by changing viticultural conditions. / Das eigene sensorische Konzept der Typizität von trockenen Rieslingweinen wurde mittels unterschiedlicher sensorischer Methoden nachgewiesen. Dies wurde insbesondere durch den Vergleich mit trockenen Weißweinen anderer Stile, vornehmlich mit trockenen Weißweinen aus Bordeaux, welche eine ausgeprägte Sauvignon blanc Stilistik aufwiesen, erzielt. Es konnte mithilfe von zwei unabhängigen sensorischen Panels, bestehend aus Weinexperten, gezeigt werden, dass die aromatische Typizität von trockenen Rieslingweinen mit „fruchtigen“ Deskriptoren, wie „Zitrus“ oder „gelbe Früchte“ zusammenhängt. Die mittels Gaschromatographie, gekoppelt mit massenspektrometrischer Detektion, (GC-MSD) gemessenen Konzentrationen flüchtiger Verbindungen wurden mit den sensorischen Daten korreliert. Hierbei wurde zum ersten Mal gezeigt, dass die Konzentrationen des hoch potenten Aromastoffes 3 Sulfanylhexanol (3SH) mit der wahrgenommenen Typizität von Rieslingweinen im engen Zusammenhang stehen. Der Einfluss weinbaulicher Bedingungen während der vegetativen Phase auf das Aroma von Weinen, einschließlich der Aromastoffvorstufen in Beeren und Most, wurde hinsichtlich der Wasserversorgung der Rebe und verschiedener Entblätterungspraktiken untersucht. Es konnte gezeigt werden, dass sich ändernde weinbauliche Bedingungen die Gehalte von Aromastoffen in den Weinen beeinflussen, insbesondere die von 3SH, Linalool und 1,1,6 Trimethyl-1,2-dihydronaphthalene (TDN). Zusätzlich hierzu wurde der Aromastoff trans-Zimtsäureethylester als möglicher beteiligter Aromastoff in der Wahrnehmung der aromatischen Typizität in Rieslingweinen identifiziert. Der hierzu angewandte vergleichende Ansatz mit trockenen Weißweinen aus Bordeaux unter Verwendung einer sensorisch geleiteten Fraktionierung mittels Hochdruck-Flüssigkeits-Chromatographie (HPLC) und anschließender zweidimensionalen Gaschromatographie gekoppelt mit olfaktorischer und hochauflösender massenspektrometrischer Detektion erwies sich als sehr gut geeignetes Werkzeug zur Dechiffrierung des Aromas von Rieslingweinen. Die Gesamtheit der Ergebnisse ermöglichte es das Wissen über die aromatische Typizität von Rieslingweinen und die beteiligten Aromastoffe zu erweitern, und einen Einblick zu bekommen wie diese Konzentrationen durch weinbauliche Faktoren beeinflusst werden.

Pevaporation is a method for separating volatile components from liquid mixtures at ambient temperatures. The paint processing industry uses Hansen solubility parameters (HSP) to indicate polymer solubility. The potential of this method to predict solvent-polymer affinity was investigated for screening potential membrane materials for the pervaporation of a model solution containing linalool and linalyl acetate (major components of lavender essential oil), in ethanol. Published HSP values were collated for various polymers, and statistically analysed to determine variations in HSP values for polymer species. An investigation of published research into pervaporation of organic/organic binary solutions separated by homogeneous membranes indicated that the solvent whose HSP value was closest to that of the polymer would preferentially permeate. This relationship did not always hold for halogenated solvents or aqueous/organic solutions. Conflicting literature regarding the relationship between solvent uptake by polymers and HSP relative energy differences was resolved using a logarithmic relationship between these two parameters. The following membranes were selected, using their HSP to indicate their potential to interact with lavender oil components: Polyamide (PA: 26.9 micro;m), Polycarbonate (PC: 20.5 micro;m), Poly(ether imide) (PEI: 29.2 micro;m), Poly(ether sulphone) (PES: 27.6 micro;m), Polyethylene (HDPE: 10 micro;m, LDPE: 13-30 micro;m), Polyimide (PI: 30.0 micro;m), Poly(methyl methacrylate) (PMMA: 50 micro;m), Polypropylene (PP: 15.9 micro;m), and Poly(tetrafluoro ethylene) (PTFE: 26.7 micro;m). The HSP (dispersive, polar hydrogen bonding components) for each membrane were calculated using the mean value obtained from swelling experiments, group contribution (calculated using Hoftyzer-Van Krevelen, Hoy and Beerbower methods), refractive indices (dispersive component), dielectric constants (polar component), and published HSP values. Pervaporation experiments investigated the effect of membrane thickness, process temperature, permeate pressure, impinging jet heights, feed flow rates and concentrations, and pre-soaking the membrane; on flow rate and selectivity in a polyethylene membrane. Membrane thickness was the dominant factor in membrane selectivity; the thinnest membranes (11.3-14.8 micro;m) had much poorer selectivity than membranes gt;24.7 micro;m. Temperatures between 22-34ordm;C, permeate pressure lt;10 kPa, impinging jet heights between 0.36-3.36 mm, feed flow rates between 541-1328 mL/min and concentrations between 1.78-6.01 % v/v of linalool and linalyl acetate in ethanol did not significantly affect selectivity. Flow rates increased with operating temperature, permeate pressure, and impinging jet heights. However, feed flow rate and concentration had no effect on membrane flux rate. Pre-soaking the membrane reduced the time to reach steady-state. Selected membranes were further investigated under standard operating conditions (permeate temperature 30ordm;C, permeate pressure lt;10 kPa, impinging jet height 1.36 mm, feed flow rate 804 mL/min and feed concentration of 5% v/v of linalool and linalyl acetate in ethanol). PMMA completely disintegrated in feed solution, and PC was too brittle to make an effective homogeneous membrane. PA, PC, PEI and PTFE had the highest efficiency (selectivity x flow rate) in their homogeneous form. However, PEI, PI and PTFE had the greatest selectivity, thus further trials should be done to improve stability and flow rates through these membranes. Pervaporation selectivity did not always follow trends predicted by HSP. Although polymers such as PA, PEI, PES, and PI preferentially permeated linalool as predicted, PC, PP and PTFE did not preferentially permeate linalyl acetate. This may have been due to the difference in size and diffusivity of these molecules (linalyl acetate, the larger molecule, did not follow the sorption selectivity predictions), or reliability of literature HSP values and those calculated by group contribution. This research shows that HSP is a good screening method for pervaporation membranes, especially where the molecules being separated are of comparable size. Polymers that have HSP close to the desired component and not to other components tend to have the best selectivity and flux characteristics. However, diffusion is an important factor, and is not completely accounted for by HSP. Recommendations for further research include: carrying out pervaporation analyses of selected polymers using pure lavender essential oil; modifying polymers to form asymmetric or composite membranes with improved permeation characteristics; and potential use of thin channel inverse gas chromatography to determine a more accurate HSP which includes diffusivity.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Lippia alba (Mill.) N. E. Brown is a native medicinal plant of South America. Known popularly as false-melissa , its leaves and roots are used in the treatment of some illnesses, including some of infective etiology. The presence of essential oils, among other metabolites with recognized antimicrobial activity, can explain its medicinal use. This paper describes the chemical analyses and antimicrobial activity of the essential oils from Lippia alba at different seasons of the year, as well as the bio-guided fractionation, the isolation and the structure elucidation of its main constituents. Lippia alba was cultivated in São Luiz Gonzaga, RS, Brazil, and its leaves were collected in 2005 and 2006 in the middle period (January, April, July and October) of each season. A voucher specimen (SMDB n° 10.050) was deposited in the herbarium of the Department of Biology, UFSM. The extraction of the essential oils was performed by the hydrodistillation method of fresh leaves. The oils were analyzed by GC-MS and their constituents were identified by the comparison of Kovat s retention indexes and mass spectra with literature data. The essential oil was fractionated by CC over silica gel for the isolation of germacrene D-4-ol and linalool, both identified by GC-MS, 13C and 1H NMR, DEPT-135, COSY and HETCOR. The antimicrobial activity of the essential oils, fractions and isolated compounds were determined by the broth microdilution method. The essential oil showed qualitative and semi-quantitative variability in relation to the seasons of the year. The yield ranged from 0.3 to 0.6%, being lower in winter and higher in summer, autumn and spring. The essential oil was composed of monoterpenoids (69 - 84%) and sesquiterpenoids (6 - 24%), being most of them oxygenated and hidrocarbonated, respectively. Moreover, the major compounds (> 15%) were not the same between the years of the analysis. The seasonal variability influenced the antimicrobial activity of the essential oils. With the exception of P. aeruginosa, all other microorganisms were susceptible to the essential oil. The most sensitive ones were S. cerevisiae (MIC and MFC of 800 μg/mL for the oil obtained in January), S. aureus (MIC of 800 μg/mL and MBC of 1600 μg/mL for the oils obtained in April and July) and K. pneumoniae (MIC and MFC of 1600 μg/mL for the oil extracted in July). In view of the antimicrobial activity, winter and summer are the best seasons for the collection and extraction of essential oils. The bio-guided fractionation of the essential oil showed that the fractions containing greater amounts of oxygenates are more active than those containing hydrocarbon compounds. Germacrene D-4-ol presented activity against S. cerevisiae (MIC and MFC of 200 μg/mL), P. zopfii (MIC and MFC of 400 μg/mL) and S. aureus (MIC and MBC of 800 μg/mL), while linalool was inactive against the tested microorganisms up to 3200 mg/mL. / Lippia alba (Mill.) N. E. Brown é uma planta medicinal nativa da América do Sul. Conhecida popularmente como falsa-melissa , suas folhas e raízes são empregadas no tratamento de várias doenças, incluindo algumas de etiologia infecciosa. A presença dos óleos essenciais, entre outros metabólitos com reconhecida atividade antimicrobiana, pode explicar o seu uso como medicinal. Este trabalho descreve a analise química e a atividade antimicrobiana dos óleos essenciais de Lippia alba nas diferentes estações do ano, bem como o fracionamento bio-guiado, o isolamento e a elucidação estrutural dos seus principais constituintes. Lippia alba foi cultivada no município de São Luiz Gonzaga, RS, Brasil, e suas partes aéreas foram coletadas no período médio de cada estação (janeiro, abril, julho e outubro) durante o ano de 2005 e 2006. Material testemunha (SMDB n° 10.050) encontra-se depositado no Herbário do Departamento de Biologia da UFSM. Para a realização dos experimentos, foram utilizadas as folhas frescas. A extração dos óleos essenciais foi realizada pelo método da hidrodestilação. Os óleos foram analisados por CG-EM e seus constituintes identificados pela comparação dos índices de retenção de Kovats e espectros de massas com dados da literatura. O óleo essencial também foi fracionado por CC sobre gel de sílica até o isolamento do germacreno D-4- ol e do linalol, ambos identificados por CG-EM, RMN 13C e 1H, DEPT-135, COSY e HETCOR. A atividade antimicrobiana dos óleos essenciais, das frações e das substâncias isoladas foi determinada pelo método de microdiluição em caldo. O óleo essencial apresentou variabilidade qualitativa e semiquantitativa em relação às estações do ano. O rendimento variou na ordem de 0,3 a 0,6%, sendo menor no inverno, e maior no verão, outono e primavera. O óleo essencial foi composto de monoterpenóides (69 - 84%) e sesquiterpenóides (6 24%), sendo a maioria deles oxigenados e hidrocarbonados, respectivamente. Os compostos majoritários (>15%) não foram os mesmos entre os diferentes anos de análise. A variabilidade sazonal, por sua vez, influenciou a atividade antimicrobiana dos óleos essenciais. Com exceção de P. aeruginosa, todos os outros microorganismos foram sensíveis ao óleo essencial de pelo menos uma estação. Os microorganismos com menor CIM foram S. cerevisiae (CIM e CFM de 800 μg/mL para o óleo obtido em janeiro), S. aureus (CIM de 800 μg/mL e CBM 1600 μg/mL para os óleos obtidos em abril e julho) e K. pneumoniae (CIM e CBM de 1600 μg/mL para o óleo extraído em julho). Tendo em vista a atividade antimicrobiana, o inverno e o verão constituem as melhores épocas para a coleta e extração dos óleos essenciais. O fracionamento bio-guiado do óleo essencial mostrou que frações contendo maiores quantidades de compostos oxigenados são mais ativas do que aquelas contendo compostos hidrocarbonados. Germacreno D-4-ol apresentou atividade contra S. cerevisiae (CIM e CFM de 200 μg/mL), P. zopfii (CIM e CFM de 400 μg/mL) e S. aureus (CIM e CBM de 800 μg/mL), ao passo que linalol foi inativo contra os microorganismos testados até a concentração de 3200 mg/mL.

Algunas plagas causan daños importantes en productos y granos almacenados, lo cual conlleva consecuentemente a pérdidas de producción y calidad en estos productos.Las principales plagas del arroz almacenado en España son, Sitophilus oryzae L. (Coleoptera Curculionidae), Rhyzopertha dominica Fabricius (Coleoptera: Bostrichidae), y Cryptolestes pusillus Schönherr (Coleoptera: Cucujidae). Las dos primeras son plagas primarias que atacan directamente el grano y resultan bastante destructivas debido a que sus larvas se alimentan y desarrollan dentro de él. C.pusillus es, sin embargo, una plaga secundaria que se beneficia de granos que ya están dañados y rotos.Actualmente, el uso de fumigantes e insecticidas de síntesis sigue siendo el principal método de lucha para controlar las plagas de almacén, si bien recientemente (dadas las continuas restricciones al uso de agroquímicos) existe un gran interés en la utilización de otras alternativas tales como el control biológico, el almacenamiento a bajas temperaturas, o los tratamientos con calor entre otros.Igualmente muchos productos obtenidos principalmente de plantas y que derivan del metabolismo secundario de las mismas ofrecen una fuente de bioinsecticidas que podrían representar una alternativa ecológica frente a los insecticidas de síntesis ya que su uso masivo e indiscriminado ha ocasionado problemas tales como la aparición de resistencias en determinadas especies de insectos frente a diferentes materias activas, desequilibrios ecológicos y problemas medioambientales sin olvidar el riesgo que entrañan para la salud humana.Con este trabajo se ha pretendido estudiar la actividad plaguicida de los aceites esenciales extraídos de tres plantas: Coriandrum sativum L. (Umbelliferae), Carum carvii L. (Umbelliferae) y Ocimum basilicum L. (Labiatae) y su posterior fraccionamiento para identificar dentro de estos aceites los compuestos químicos responsables de esta actividad insecticida sobre tres plagas de almacén de arroz (S. oryzae, R.dominica y C.pusillus).Del estudio de estos monoterpenoides, linalol, S-carvona y estragol resultaron tener una alta actividad insecticida sobre estas plagas. Sin embargo el E-anetol fue más selectivo para R.dominica y C.pusillus, así como el limoneno, γ-terpineno, geraniol y eucaliptol sólo resultaron activos frente a C.pusillus.Algunos monoterpenoides podrían actuar de sinergistas potenciando la actividad de otros, como podría ser el caso del alcanfor, acetato de geranilo y E-anetol con linalol en R.dominica y C.pusillus, o el caso del metoxicinamaldehido, p-anisaldehido y linalol que pueden tener efecto sinergista sobre el estragol.Este trabajo también abarcó el estudio de un posible modo de acción de estos monoterpenoides, la inhibición de la acetilcolinesterasa, para alcanzar un mayor entendimiento del comportamiento de estas sustancias en el interior del insecto.Así se pudo observar como la mayoría de monoterpenoides estudiados inhibían en cierta medida esta enzima, siendo fenchona, S-carvona y linalol los monoterpenoides que mayor inhibición originaron.Del mismo modo se observó como fenchona, γ-terpineno, geraniol y linalol inhibían competitivamente la acetilcolinesterasa, mientras que S-carvona, estragol y alcanfor producían una inhibición mixta para esta enzima.Sin embargo no se observó inhibición de la acetilcolinesterasa por parte del E-anetol a las concentraciones de monoterpenoides ensayadas.Para completar este trabajo se examinó de igual forma la capacidad que tenían estos bioinsecticidas de generar resistencia en estas tres plagas así como el mecanismo de resistencia implicado en el desarrollo de este proceso. Para ello se fueron seleccionando las poblaciones de insectos mediante la aplicación de los diferentes monoterpenoides a dosis crecientes durante 7 generaciones. De este modo se pudo calcular el factor de resistencia en cada una de las plagas y para cada uno de los monoterpenoides comparando las concentraciones letales 50 de las poblaciones seleccionadas con las poblaciones iniciales (sensibles).A continuación se analizaron tres posibles sistemas de detoxificación enzimáticos gracias al uso de sinergistas para estudiar el mecanismo de resistencia que podría estar involucrado.De tal forma se observó como estos monoterpenoides inducían lentamente resistencias resultando ventajosos en un futuro para el control de estas plagas.Esta Tesis ha englobado un estudio íntegro y profundo de estos insecticidas ecológicos desde la extracción de los aceites esenciales, seguido de la identificación de compuestos puros (CG-EM) con actividad insecticida junto con el estudio de un modo de acción de estos insecticidas, la selección de resistencia en las poblaciones de insectos y los posibles mecanismos de resistencia que pudieran estar implicados en este proceso. / Some pests cause serious damage to stored grains and stored products and consequently production and quality losses in these products.The rice weevil, Sitophilus oryzae L. (Coleoptera: Curculionidae), the lesser grain borer, Rhyzopertha dominica Fabricius (Coleoptera: Bostrichidae) and Cryptolestes pusillus Schönherr (Coleoptera: Cucujidae) were the main damaging pests found in stored rice in Spain.S.oryzae and R.dominica are primary pests attacking directly the intact grain and are quite destructive because their larvi feed and develop inside the grain whereas C.pusillus is a secondary pest which benefits from grains previously damaged.At the present time, organic synthetic pesticides are still the main method to control stored grain pests, however, recently (due to restriction in agrochemicals use) there is a great interest in using other altenatives such as biological control, storage at low-temperatures, or heat treatment.Likewise, many products obtained mainly from plants and derived from secondary metabolism have insecticidal activity against insects, such as monoterpenoids, which present a broad variety of bioinsecticide products which could turn out to be an ecologic alternative to synthetic pesticides since the majority of alternative products are not harmful for the human healthy and they become less environmentally damaging, exhibiting a low impact on the environment.In addition, it cannot be ignored the different difficulties related to resistance due to several active compounds from organic pesticides found in some species of insects.With this work, we have considered remarkable to study the insecticide activity of essential oils extracted from three plants: Coriandrum sativum L. (Umbelliferae), Carum carvii L. (Umbelliferae) and Ocimum basilicum L. (Labiatae) and carry out a bioassay-guided fractionation of their essential oils to identify which compounds were responsible for the volatile toxicity shown on three stored rice pests (S. oryzae, R. dominica and C. pusillus).Linalool, S-carvone and estragole turned out to have a high insecticide activity on these pests. Nevertheless E-anetol was more selective to R. dominica and C. pusillus, being only active on C. pusillus limonene, γ-terpinene, geraniol and eucalyptol.Some monoterpenoids were found as synergists, increasing the activity of the other ones, like for instance, camphor, geranyl acetate and E-anethole with linalool in R.dominica and C.pusillus or metoxycinnamaldehyde, p-anysaldehyde and linalool which could activate to estragole.In this work, the inhibition of acetylcholinesterase as a posible mode of action was studied as well, to reach a clear understanding about the action of these products inside the insects.The majority of monoterpenoids inhibited the enzyme acetylcholinesterase being fenchone, S-carvone and linalool the monoterpenoids that produced a higher inhibition.Furthermore, it was observed how fenchone, γ-terpinene, geraniol and linalool showed a competitive inhibition whereas S-carvone, estragole and camphor produced a mixed inhibition for this enzyme. However the enzyme acetylcholinesterase was not inhibited by E-anethole.To finish up this work, the selection for monoterpenoid resistance on these pests as well as the metabolic mechanisms implicated was studied.The resistant strains were selected from susceptible insect populations and survivors were reared separately for each monoterpenoid and successive generations were treated with higher concentrations. These populations were selected until seven times.As a result we could calculate the resistance factor on each pest (comparing lethal concentration 50 values of susceptible and resistant strains).Next, three enzymatic systems detoxifying these monoterpenoids were analysed to study the metabolic mechanism implicated.In this way we could realize that all monoterpenoids induced resistance slowly, concluding that these pesticides will be appropriated to control these pests in the future.This Thesis has concerned a study in depth about ecological insecticides from extraction of essential oils, identification of compounds (GC-MS) with insecticide activity, mode of action and study of resistance and mechanism of insecticide resistance involved in this process.