O gênero Eupatorium L. (Asteraceae) apresenta muitas espécies comumente encontradas no Rio Grande do Sul e se caracteriza pela presença de óleos voláteis e outros derivados terpenoídicos entre seus metabólitos secundários. Neste trabalho, o óleo volátil de oito espécies de Eupatorium nativas do estado foi extraído por hidrodestilação em aparelho tipo Clevenger e sua composição química foi avaliada por cromatografia gasosa acoplada a espectrometria de massas (CG/EM). As espécies analisadas foram: E. inulifolium Kunth, E. polystachyum DC, E. picturatum Malme, E. tremulum Hook. & Arn, E. ivifolium L., E. laevigatum Lam., E. casarettoi (B.L.Rob.) Steyerm., E. itatiayense Hieron. e E. gaudichaudianum DC. Os óleos voláteis de partes aéreas de E. tremulum analisados antes, durante e após a floração, aplicando-se análises multivariadas para identificar compostos com variação significativa em cada estágio de desenvolvimento da espécie. Os extratos diclorometano das mesmas espécies foram avaliados na busca de lactonas sesquiterpênicas utilizando como metodologia a mensuração da absorção característica de lactonas no espectro Infravermelho e aplicação de métodos de extração específicos para essa classe de compostos. Foi realizado fracionamento dos óleos voláteis em cromatografia em coluna aberta (CC), cromatografia flash (CC flash) e cromatografia líquida de média pressão (MPLC) com monitoramento das frações por CG/EM para o isolamento de compostos não identificados apenas por seu índice de retenção linear e espectro de massas. Utilizando essa abordagem foi isolado e identificado por seus dados espectrais de RMN um furanossesquiterpeno, 3-oxoverboccidentafurano, descrito pela primeira vez em Eupatorium. Também foi realizado o fracionamento de extratos diclorometano (E. casaretoi e E. inulifolium) em CC flash e MPLC, com monitoramento das frações por CG/EM e cromatografia líquida de alta eficiência acoplada a detector de arranjo de diodos (HPLC-DAD) na tentativa de isolamento de lactonas sesquiterpênicas. Um composto foi isolado e sua completa elucidação estrutural está em andamento. Considerando as dificuldades encontradas no processo de elucidação estrutural da molécula isolada do extrato de E. inulifolium, mesmo com a abundância de dados de ressonância magnética nuclear (RMN) 13C disponíveis para consulta, e que não foi encontrada uma base digital dedicada a lactonas sesquiterpênicas reunindo esses dados em uma plataforma facilmente acessível e utilizável, foi realizada a transposição de uma base de dados da literatura para um formato digital amigável (baseada em Excel®), com a visualização e comparação de dados facilitadas graças à utilização de formas gráficas em lugar das formas tabulares de apresentação dos espectros comuns na literatura. / The genus Eupatorium L. (Asteraceae) contains several species that are common in Rio Grande do Sul State and it is characterized by essential oils and other terpenic derivatives among its secondary metabolites. Through this survey, it was evaluated the chemical composition of the essential oil from eight species of Eupatorium natives to the State and their chemical composition was analysed by gas chromatogaphy coupled to mass spectrometry. The following species were studied: E. inulifolium Kunth, E. polystachyum DC, E. picturatum Malme, E. tremulum Hook. & Arn, E. ivifolium L., E. laevigatum Lam., E. casarettoi (B.L.Rob.) Steyerm., E. itatiayense Hieron. and E. gaudichaudianum DC. Volatile oils from aereal parts of E. tremulum were analysed before, during and after blooming. Multivariate analyses were employed in order to identify compounds showing significant variation between each developmental stage. Dichloromethane extracts of these same species were evaluated for sesquiterpene lactones by measuring the typical infrared absorption for lactones and applying extraction methods directed to this class of compounds. The essential oils were fractionated by CC, CC flash and MPLC, with fractions being monitored by GC/MS for isolation of unknown compounds, whose retention index and mass spectrum were not enough for identification. Using this approach a furansesquiterpene, 3-oxo-verboccidentafuran, described for the first time for Eupatorium, was isolated and identified based on NMR spectral data. Dichloromethane extracts (E. casarettoi and E. inulifolium) were also fractionated by CC flash and MPLC, fractions were monitored by GC/MS and HPLC-DAD aiming the isolation of sesquiterpene lactones One compound was isolated and its structural elucidation is ongoing. Considering difficulties faced in the process of structural elucidation of the compound isolated from the extract of E. inulifolium, even with plenty of NMR 13C data available for search, and considering that no digital database dedicated to sesquiterpene lactones was found, that gathers these data in an easily accessible and user friendly platform, we performed the transposition of a bibliographic database to a more user friendly digital format (baseada em Excel®), with enhanced data visualization and comparison, thanks to the use of graphical rather than the tabular spectral presentation usually found in the literature.

Athrixia phylicoides (bush tea) is one of many plants from the Asteraceae family used as a traditional herbal medicine. With very few cultivated plants, natural growing plants currently serve as the main resource for plant material. The plant is not yet commercialised and its medicinal value is known and used only by a few people. With the long term aim at commercial scale propagation, this study consists of three parts. Firstly we developed a protocol for in vitro propagation of A. phylicoides. Secondly, the ultrastructure and morphology of leaves were studied microscopically and thirdly, comparisons were made between in vitro and ex vitro grown plants. Nodal segments of greenhouse plants were used to establish cultures. Better growth and less wilting was recorded on explants surface sterilised with NaOCl compared to Ca(OCl)2 after establishment. The addition of growth regulators IBA (indole-3-butyric acid) and BAP (6-benzylaminopurine) to the culture medium did not seem to affect the growth response of explants during the multiplication phase. Hyperhydricity was a problem throughout our trials. The development of hyperhydricity symptoms seems to be related to seasonal changes in the stock plant material used to initiate cultures, rather than the composition of growth medium or growth room temperatures. The occurrences of hyperhydricity symptoms were inconsistent and unpredictable. A rooting medium with added BAP and decreased sucrose levels resulted in a higher rooting percentage compared to the control medium, free from BAP and with a higher sucrose concentration, which yielded no rooting. However, in another experiment, in vitro rooting occurred spontaneously after subdividing and transfer of microshoots to fresh control medium. The addition of GA3 to the establishment medium (but not to the subsequently used multiplication and rooting media) yielded a slightly higher percentage of rooting. However, cultures initially established on GA3 medium yielded fewer roots per explant and roots were shorter than those of explants established on hormone free medium. The medicinal properties of plants are often linked to the production of essential oils. We hypothesised that the medicinal value of A. phylicoides can be linked to the production of the aromatic essential oils released by leaves. A microscopic study of leaves provided some preliminary insight of the mechanisms involved in the production of medicinally active products. Electron- and light microscopic examination of leaves were used to identify and study structures that are apparently involved in the production and secretion of essential oils. Two types of trichomes were identifyed – nonglandular and glandular trichomes. These glandular trichomes are multicellular with a subcuticular storage space and are present only on the adaxial surface of leaves. In the case of medicinal plants, it is essential that the medicinal properties of the plant are not altered by the method of propagation. This was our motivation for comparing the morphology and ultrastructure of leaves of plants that were grown in their natural environment to that of plants grown in vitro. Leaf surfaces of in vitro grown plants were smaller and the number of glandular trichomes per surface area was less on in vitro grown plants. There were no noticeable changes in the morphology of glandular trichomes. / Dissertation (MSc(Agric))--University of Pretoria, 2009. / Plant Production and Soil Science / unrestricted

Experiments were conducted to study variation in the amount of podophyllotoxin in J. virginiana across nine –physiographic regions of Mississippi. The aim was bioprospection of podophyllotoxin from J. virginiana across Mississippi. Experiments were also conducted to utilize biproducts (i.e. needles) as a source of podophyllotoxin and essential oil. Dual extraction of both from the same plant material was obtained in order to develop economic protocol for industrial utilization. Three different experiments were conducted for the bioprospection and byproduct utilization of J.virginana. Experiment I: Study of variation in the amount of podophyllotoxin across nine physiographic regions and the study of contributing factors for the variability. Experiment II: Explore different plant parts as source of podophyllotoxin Experiment III: Establish a dual extraction protocol for extraction of podophyllotoxin and essential oil from J. virginiana.

Essential oils are natural products used to flavor food and beverages. With the increase in nutrition conscious consumers, manufacturers of food additives and food products are faced with the challenge of making healthy alternatives. In particular, food products going to market with label claims stating reductions in sugar and salt, organic certified, organic compliant, and all natural; moreover the ingredients used in flavors must meet these label claims as well. More often than not, the challenge in using ingredients that follow these requirements is the pricing, the sourcing and the variability among those sources. Variability is common in the ingredients coming from nature such as fruits and plants because the area of cultivation can vary by the soil at the sight of planting and/or the climate in the region. Pricing is also problematic in naturally grown ingredients because it is a matter of supply and demand. Stock could be depleted from natural disasters, disease carrying pest(s), pests that consume the crop, and/or other causes for scarce supply of crop(s). Essential oils are natural byproducts of fruit, peels, and leaves from plants that contribute to flavor formulae for a large variety of food products. Because the essential oils are a crop based commodity, every variety has inherent differences based on the growing conditions and their ripening stages [1]. Nevertheless, each type of oil has marker chemicals that make up the majority of its composition; these marker chemicals have the tendency to degrade over time based on their interaction with light, oxygen exposure, and temperature. For companies that manufacture flavorings, understanding the variability among sources of essential oils as well as the possible degradants of essential oils is valuable information to obtain because it is possible the variants and degradants will negatively impact the flavor profile. Flavor is without question the most important attribute of the food we consume and by default stability of said flavor(s) need to be understood [30]. The content in this dissertation involves the stability analysis of a common essential oil, Oil Mandarin Italian Select, from Citrus Reticulata Blanco. It has known off notes that form from unknown causes. Most common is the plastic note that has formed in carbonated products like soda. Studying this particular essential oil in various conditions is intended to shed light on what those degradants are and under which conditions they form to give mandarin oil an off-note when applied to high acid and carbonated beverage applications. Once the note is reproduced, a correlation between analytical data and sensory interpretation of the oil will be developed. Mandarin essential oil being in the Citrus genus is traditionally analyzed via gas chromatography (GC) because of the high quantities of volatile constituents that give an oil high aroma activity. The volatile fraction of mandarin oil to be studied includes stability of methyl-N-methylanthranilate (MNMA), a major component giving mandarin its distinct grapey character, as well as gamma terpinene, thymol, sinensal, alpha pinene, beta pinene, myrcene, para cymene, alpha terpineol, and beta caryophyllene. Each of these ten compounds contributes to the unique flavor profile of mandarin oils when compared to orange and tangerine essential oils [1]. It was the common knowledge that para cymene can be perceived as rancid in aroma and the many interconversions the terpenes make that cause para cymene formation in Citrus oils, which made monitoring the changes of this chemical in the three stability environments crucial. Attention is being paid to para-cymene, as a specific marker of degradation in Citrus. The data obtained from the applied stability studies were challenging to understand as the marker chemicals are volatile and sensitive to chemical change. In this work the chemical changes and trends were analyzed under various storage conditions. Significant statistical analyses were employed to help define criteria of usability. The analyses were required because of natural variants and apparent inconsistencies of the data. Dixon Q Test and the Z Test were applied to determine outliers. Additionally, the Bland Altman method was applied to compare storage conditions and to determine if this statistical approach could be used to define significant changes in the marker chemical stability. The Bland Altman plots suggest that each component met the statistical limits of agreement, meaning the samplings were not significantly changing, statistically speaking. A final approach to assess the analytical data of the mandarin oil for significant change was the mass balance of each marker chemical from week 0 to week 24. Instrumental fluctuations have an acceptable range of +/- 20% in the industry; hence, a significant change criterion for a chemical in the mass balance must be one that exceeded +/- 20%. Unlike classical statistic methods, the mass balance was indicative that significant change had occurred to the compounds in the three studies. Upon sensory analysis of the oil samples, display of plastic note, oxidation, and overall loss of characteristic mandarin notes, the mass balance was found to correlate best to the significant change detected by sensory evaluation of the oil samplings. Due to the inadequate number of validated methods on Citrus essential oil research and the absence of large groupings of terpenes validated in a unified methodology, reconciliation of mass balance is an underutilized method of assessment in the literature. As a final assessment of the GC method validated, a product containing the selected mandarin oil was analyzed to evaluate the ability of the method to separate the oil components within a significantly more complicated matrix than the initial samples. The method was successful though not all marker chemicals were detected due to their low formulation concentration being below the level of detection of the method. This should not be seen as a failure of the method. For the major components of the essential oil studied, the method was quantitatively successful, meeting industry requirements. / Chemistry

As a response of the need for a natural antimicrobial to replace sodium benzoate's use as a preservative in beverages, twenty eight compounds known to have antimicrobial activity were evaluated to quantify their solubility. Twenty three of the compounds evaluated are components of plant essential oils and the remaining five compounds are alkyl esters of para-hydroxybenzoic acid. The test compounds were evaluated for aqueous solubility as well as their solubility in an acid-based beverage mixture. The compounds were found to be practically insoluble (< 100 mg/L), very slightly soluble (100 mg/L – 1,000 mg/L) or slightly soluble (1,000 mg/L to 10,000 mg/L). o-Methoxycinnamaldehyde, trans,trans-2,4-decadienal, cinnamic acid, and citronellol were complexed with α- and β-cyclodextrin and evaluated through phase solubility analyses. The complexes formed showed improved aqueous solubility for all compounds. Complexation with α-CD resulted in an increase of aqueous solubility of o-methoxycinnamaldehyde by 10-fold, trans,trans-2,4-decadienal by 3.2-fold, cinnamic acid by 6.3-fold, and citronellol by 8-fold. In addition, complexation with β-CD resulted in an increase of aqueous solubility of o-methoxycinnamaldehyde by 1.6-fold, trans,trans-2,4-decadienal by 3.1-fold, cinnamic acid by 1.7-fold, and citronellol by 1.6- fold. The storage stability of the α-CD complexes of o-methoxycinnamaldehyde trans,trans-2,4-decadienal and citronellol were evaluated for 7 days in an acid-based beverage solution by SPME GC-MS. The complexes exhibited varying levels of degradation throughout the duration of the study all. The concentration of o-methoxycinnamaldehyde detected by SPME GC-MS decreased by 61.7%. Similarly, the concentration of trans,trans-2,4-decadienal and that of citronellol decreased by 62.7% and 43% respectively. Additionally, a comparison by UV/Vis of the storage stability of the complexes stored in glass and PET containers was performed. The storage stability comparison proved that absorption into the PET polymer membrane did not occur. / Master of Science

O uso de óleos essenciais na indústria de fragrância tem crescido a cada ano. A indústria de cosméticos, em geral, tem buscado este tipo de ingrediente com o objetivo de agregar atributos únicos e melhores aos seus produtos e assim entregar, ao consumidor final, diferenciais que poderão valorizar suas formulações, além de simplesmente perfumar. O benefício de usar óleos naturais ou acordes de seus principais componentes em formulações de fragrâncias é que estes se tornam composições mais ricas, e em muitos casos, capazes de proporcionar perfumação prolongada ao produto, quando comparados àqueles sintetizados. A presença de compostos variados nos óleos, como terpenos e resinas, ajudam a promover singularidades à fragrância e até mesmo servem como inspiração às criações de produtos com descrições olfativas de produtos naturais. Croton tricolor (Euphorbiaceae), segundo avaliação inicial de um perfumista, apresentou excelentes características olfativas, o que facilitou seu uso nos estudos. A análise do óleo essencial de C. tricolor por cromatografia à gás com detector de ionização de chamas (CG/DIC) e cromatografia à gás acoplada à espectrometria de massas (CG/EM) permitiu quantificar e identificar seus principais marcadores, bem como utilizar estas informações para construção de acorde olfativo, reproduzindo o odor do produto natural. O óleo essencial de C. tricolor apresentou como principais componentes o biciclogermacreno (15,4%), espatulenol (10,5%) e o alfa-pineno (6,8%). A partir destes dados foi construído um acorde aromático que reproduziu o odor original, contendo alfa-pineno (7%), álcool fenetílico (5%), óleo de cedro (5%), óleo de laranja (0,5%), limoneno (1%), eucaliptol (0,5%) e nerolidol (0,3%). Os compostos majoritários, biciclogermacreno e espatulenol, não foram adicionados por não serem disponíveis comercialmente, por isso, usaram-se outras matérias-primas semelhantes em perfil olfativo e ofertadas no mercado, como foi o caso do uso de álcool fenetílico, óleo de cedro e óleo de laranja na criação do acorde. Os resultados evidenciaram que a criação de acordes é um método economicamente viável para a reprodução de odores naturais que podem ser utilizados em formulações de fragrâncias, além de ajudar a resolver problemas na produção e comercialização de óleos essenciais como, sazonalidade e reprodutibilidade. / The use of essential oils in the fragrance industry has grown every year. The cosmetics industry has generally sought this type of ingredient, with the aim of providing its products with differentiated benefits (unique and better attributes), and thus delivering to the final consumer a differential that can enhance their formulations, besides simply perfuming their products (theirs). The benefit of using natural oils or accords of their major components in fragrance formulations makes them richer, and in many cases capable of providing prolonged perfuming to the final product when compared to synthesized products. The presence of varied compounds in the oils, such as terpenes or (and) resins, helps to promote differentiated fragrance characteristics (singularities/peculiarities to the fragrance), or (and) even to serve as inspiration for product creations with olfactory descriptions of natural products. Croton tricolor (Euphorbiaceae), accordingly to an initial evaluation of a perfumer, presented excellent olfactory characteristics, which facilitated its use in the studies. The analysis of the C. tricolor essential oil by gas chromatography with flame ionization detector (GC/FID) and gas chromatography coupled to mass spectrometry (GC/MS) allowed to quantify and identify their main markers, as well as to use this information to construct an olfactory accord that reproduces the odor of natural products. The C. tricolor essential oil had as main components bicyclogermacrene (15.4%), spathulenol (10.5%) and alpha pinene (6.8%). From these data, an aromatic accord was created reproducing the original odor containing alpha pinene (7%), phenethyl alcohol (5%), cedar oil (5%), orange oil (0.5%), limonene (1%), eucalyptol (0.5%) and nerolidol (0.3%). Some of the major compounds, bicyclogermacrene and spatulenol, could not be added because they were not commercially available. In these cases (therefore), other products (raw materials) with similar olfactory characteristics (olfactory profile) and commercially viable (offered on the market) were used, such as the use of phenethyl alcohol, cedar oil and orange oil in the accord creation. The results showed that accord creation is an economically viable method for the reproduction of natural odors that can be used in fragrance formulations and may help to solve problems that exist in the production and commercialization of essential oils such as production seasonality and reproducibility.

No presente estudo o óleo essencial da Aniba duckei Kostermans (Pau Rosa) cultivada em ambiente natural foi extraído quantitativamente, por hidrodestilação, de folhas secas e galhos finos. Foram determinadas propriedades físicas e químicas e o rendimento, incluindo o estudo do tempo de extração, da extração com força iônica controlada e com várias constantes dielétricas. Promoveu-se a acetilação do óleo essencial para obtenção do acetato de linalila e verificou-se aspectos cinéticos da reação. As técnicas de espectroscopia no ultravioleta, infravermelho, de massas e ressonância magnética nuclear de 13C foram empregadas para a identificação do componente mais abundante e seu derivado, pela modificação química. Usou-se a cromatografia gasosa para a quantificação, pelo método do padrão externo, do principal componente e de seu derivado obtido por acetilação. Os valores das propriedades físicas determinadas estão concordantes com aqueles encontrados nas espécies nativas. Os rendimentos médios foram de 1,3 e 1,2% para folhas secas e galhos finos, respectivamente. Os espectros no IV, RMN 13 C e EM confirmaram a presença majoritária do linalol. A concentração deste foi de 72,3 e 78,1 % no óleo essencial extraído de folhas secas e galhos finos, respectivamente. No óleo essencial adquirido da Destilaria de Presidente Figueiredo/AM (extraído da madeira) a concentração foi de 90,3 %. Obteve-se pela acetilação do óleo essencial da destilaria 49,0 % de acetato de linalila, fornecendo 54% de rendimento. Os resultados obtidos mostraram uma grande possibilidade para a exploração do óleo de folhas e galhos finos, com a incisão de alto valor agregado neste produto. / In the present study the essential oil of the Aniba duckei Kostermans (Pau Rosa) cultivated in natural environment was extracted quantitatively, by hidrodistillation, from dry leaves and small branches. Physical and chemical properties and the yield of essential oil were determined. The study of the time of extraction, extraction with controlled ionic strength and with several dieletrics constant were done. Acetilation of the essential oil was promoted to obtain the linalyl acetate and kinetic aspects of the reaction were verified. The spectroscopic techniques in the ultraviolet, infrared, of masses and nuclear magnetic resonance of 13C were used for the identification of the major component and its derivatives. The gas chromatography was used for the quantification, by the method of the external standard, of the major component and its derived obtained by acetilation. The values of the physical properties are according to ones found in the native species. The medium yield were 1,3 and 1,2% for dry leaves and small branches, respectively. IR, RMN 13C and MS spectra confirmed the majority presence of the linalool. Its concentration was 72,3 and 78,1% in the extracted essential oil of dry leaves and small branches, respectively. In the essential oil acquired of the Distillery in Presidente Figueiredo/AM (extracted from wood) the concentration was 90,3%. The linalyl acetate obtained by acetilation of the essential oil from distillery was 49,0%, supplying 54% of yield. The obtained results showed a great possibility for the exploration of the oil from leaves and small branches, with the incision of high value joined in this product.

A planta Syzygium aromaticum L. produz os botões secos do cravo da índia, do qual se extrai o óleo essencial. O componente majoritário deste óleo é o eugenol, cujas algumas de suas propriedades farmacológicas são atividades fungicida, bactericida, anestésico, entre outros. No entanto, o eugenol (EUG) foi convertido a acetato de eugenila (AE) pela adição do íon acetato, na ligação do grupo hidroxila. Esta modificação foi realizada devido aos problemas relacionados à solubilidade, o que estabelece uma difícil relação com sua biodisponibilidade, e então, limitando sua utilização em preparações farmacêuticas. Em seguida, uma mistura eutética foi preparada empregando o acetato de eugenila como princípio ativo, e o polietilenoglicol 3350 (PEG) como carreador hidrofílico. O diagrama de fases apontou a mistura eutética com composição de 80% de AE e 20% PEG. As características térmicas no ponto eutético foram de 21,33&deg; C para temperatura de pico e 5,71Jg-1 a energia envolvida no processo de fusão. Dessa maneira, a taxa de permeabilidade e a solubilidade do acetato de eugenila serão aprimoradas, contribuindo para o aumento de sua biodisponibilidade e assim, viabilizando sua utilização em formulações farmacêuticas. / The Syzygium aromaticum L. plant produces the cloves buds which are dried. From these dried buds the essential oil is extracted. The major compound is eugenol. Some of eugenol\'s pharmacological properties include fungicidal, bactericidal, anesthetic, and other activities. However, eugenol (EUG) was modified to eugenyl acetate (EA) by adding the acetate group to the bound hydroxyl. The modification was performed in order to eliminate the solubility problems which cause a difficult relationship with bioavailability. The solubility problem restrains its use in pharmaceutical formulations. A eutectic mixture was prepared using the eugenyl acetate as the principal active ingredient and a polyethyleneglycol 3350 (PEG) as the hydrophilic carrier. The phase diagram demonstrated that the eutectic mixture occurred with 80% EA and 20% PEG composition. The thermal characteristics at the eutectic point were 21.33&deg;C for the peak temperature and 5.71Jg-1 for the energy of fusion. Thus, the rate of permeability and solubility of eugenyl acetate will be enhanced. This will contribute to an increase in its bioavailability, thus promoting its use in pharmaceutical formulations.

A Lippia sidoides é planta largamente difundida nas práticas da medicina popular no Brasil, sendo utilizada como anti-séptico de uso tópico, fato justificável pela presença de timol. Como estratégia para a obtenção de novos produtos com atividade antimicrobiana, a partir de fonte vegetal, o presente estudo visou o desenvolvimento tecnológico de extratos secos padronizados e a encapsulação do óleo essencial dessa planta, através da técnica de secagem por atomização (spray drying). Para obtenção dos extratos secos padronizados, utilizou-se metodologia capaz de avaliar a qualidade da matéria-prima vegetal, das ações de transformação, dos produtos intermediários e do produto final. A solução extrativa foi obtida a partir das folhas secas e moídas dessa planta, empregando-se o método de extração por maceração dinâmica, avaliando-se alguns fatores que influenciavam sua eficiência. A solução extrativa selecionada foi submetida à secagem por atomização, investigando-se a combinação de adjuvantes tecnológicos sobre o desempenho do equipamento e sobre características físico-químicas do produto obtido. Além disso, avaliou-se a atividade antifúngica in vitro dos extratos secos, confirmando-se o efeito antimicrobiano dos mesmos. Para a encapsulação do óleo essencial de Lippia sidoides, utilizou-se a técnica de spray drying (método físico) e a inclusão molecular com -ciclodextrina (método químico) com subseqüente secagem por spray drying. Na microencapsulação por spray drying foi utilizado um conteúdo de óleo de 20 e 25% em relação ao material de parede. Como material de parede foram empregadas diferentes proporções de maltodextrina DE 10 e goma-arábica, sendo as emulsões de alimentação atomizadas até à concentração de 60% de sólidos totais. A eficiência da encapsulação foi avaliada através da quantidade específica de óleo essencial encapsulado nas micropartículas, sendo obtido valor máximo de 65%, dependente das condições experimentais empregadas. Observou-se existir teor de sólidos ótimo (50%) e relação entre o aumento da retenção de óleo total e as maiores concentrações de goma-arábica nas emulsões de alimentação, sendo que todas as micropartículas demonstraram atividade antifúngica. A inclusão molecular do óleo essencial, através de sua complexação com moléculas de -ciclodextrina exibiu eficiência de encapsulação de até 70%. As diferentes proporções de óleo essencial e -ciclodextrina testadas influenciaram esses resultados, sendo que menores retenções foram observadas quando maiores quantidades de óleo foram adicionadas às suspensões de alimentação. Pelas análises térmicas foi possível demonstrar a mais alta estabilidade térmica dos produtos encapsulados (micropartícula/complexo de inclusão) comparados ao óleo essencial original. Os dados adquiridos durante os estudos de desenvolvimento de produtos secos, a partir de Lippia sidoides, indicaram o potencial desses como agentes antimicrobianos naturais para fins medicinais, representando assim, alternativa para o aproveitamento da espécie vegetal pelo setor farmacêutico. / Lippia sidoides is an aromatic shrub widely used in a folk medicine in Brazil as local antiseptic, justified by thymol presence as the major constituent of its essential oil. As strategy for production of new antifungal herbal products, the present study aimed the technological development of Lippia sidoides standardized spray dried extracts as well as the encapsulation of its essential oil by spray drying/ molecular inclusion. In order to obtain the standardized dried extract, process control parameters of manufacturing operations were established and they were continuously tracked to provide reproducibility from batch-to-batch and to assure productsquality. Extraction (maceration) from Lippia sidoides leaves was used to produce thymol-containing liquid extracts. Optimal extraction conditions were determined and the selected extractive solution was spray dried. Effect of different carrier ratios on physicochemical and antifungal properties of dried extracts was evaluated. Lippia sidoides dried extract showed an important antifungal effect against the tested strains. Lippia sidoides essential encapsulation was carried out by spray drying technique (physical method) and molecular inclusion within -cyclodextrin (chemical method) followed by spray drying of the slurries in order to produce inclusion complex in a powder form. For microencapsulation by spray drying, maltodextrin DE 10 and gum arabic in different were used as carrier. Content of essential oil related to carrier was 20 and 25% in weight and the emulsions were atomized from 30% up to 60% of total solid concentration. Encapsulation efficiency was estimated through determination of the content of essential oil in the microcapsules and a maximum value obtained was 65%, depending on experimental parameters adopted. An optimal solid content of the encapsulating composition (50%) was observed. The increase of gum arabic amount in the infeed emulsion was related to the increase in the total oil retention in the microparticles. Antifungal activities of microparticles were evaluated, evidencing their potential as important antifungal agent. For inclusion complex formation between essential oil and -cyclodextrin, the encapsulation efficiency was up to 70%. The entrapment ability was influenced by the different essential oil: -cyclodextrin ratios tested. A decreasing tendency in the total oil content was observed, when the initial amount of added oil was increased. The greater thermal stability of the encapsulated products (microparticles/ inclusion complexes) in comparison to the original oil was confirmed by thermal analysis. The finding acquired during the development of Lippia sidoides dried products indicated their potential as natural antimicrobial agent for medicinal propose and provided evidences which support the use of such plant specimen by pharmaceutical industry.

A 1,3-dimetilamilamina (DMAA) é um estimulante que passou a ser adicionada aos suplementos alimentares e compostos emagrecedores a partir de 2006, sendo amplamente consumida por atletas e militares americanos. No entanto, após relatos de toxicidade a DMAA foi proibida por agências regulatórias do Brasil e Estados Unidos. Porém, mesmo após a sua proibição, a DMAA ainda pode ser encontrada em suplementos alimentares. A sua origem foi relacionada ao óleo essencial de Pelargonium graveolens, e, no entanto, inúmeros autores questionaram os resultados originais e a sua origem natural. Adicionalmente, os óleos essenciais de espécies de Pelargonium tiveram a sua atividade antimicrobiana reportada frente a bactérias e fungos. Assim, considerando os aspectos abordados, este trabalho teve como objetivo determinar a presença de DMAA nos óleos essenciais de Pelargonium spp. por GC-MS, DART-MS/MS e LC-MS/MS; assim como nas folhas das mesmas espécies, utilizando a extração por headspace, previamente otimizada, seguida de análise por GC-MS. Também se propôs a investigar a atividade antifúngica dos óleos essenciais de P. graveolens de diferentes origens e desenvolver uma formulação contendo uma nanoemulsão do óleo para o tratamento de candidíase vaginal. Por fim, teve como objetivo desenvolver metodologia de screening para avaliar a presença de DMAA e outros estimulantes em suplementos alimentares apreendidos, através de DART-MS/MS. Os resultados revelaram que a DMAA não está presente nos óleos essenciais de diferentes espécies de Pelargonium spp. obtidos por hidrodestilação, do Rio Grande do Sul. Após a otimização através de desenho experimental, a técnica de headspace provou ser eficaz na extração dos constituintes voláteis presentes nas folhas e, no entanto, a DMAA não foi detectada, assim como nos óleos essenciais comerciais de P. graveolens do Brasil, China, Egito, África do Sul, Albânia e Ilhas Reunião. Os óleos essenciais apresentaram atividade antifúngica frente às cinco espécies de Candida. Ainda, este efeito antifúngico apresentou melhores resultados com a nanoformulação contendo o óleo essencial. A análise de screening por DART-MS/MS se mostrou eficaz na detecção de DMAA, efedrina, sinefrina, cafeína, sibutramina e metilfenidato, em amostras de suplementos alimentares apreendidos, apresentando resultados positivos para todos os estimulantes. Com base nos resultados obtidos e nos objetivos propostos, verificou-se que mesmo após a 12 utilização de três técnicas analíticas distintas e uma nova alternativa para extração dos constituintes voláteis, a DMAA não foi econtrada nos óleos essenciais e nas folhas das espécies de Pelargonium, corroborando com outros estudos realizados, e indicando que a sua origem não é natural nestas espécies. A formulação final contendo a nanoemulsão com o óleo essencial apresentou atividade antifúngica superior a do óleo essencial livre. As análises das amostras apreendidas mostraram que mesmo após a sua proibição pelas agências regulatórias, os suplementos contendo DMAA e outros estimulantes ainda são comercializados, representando um grande risco para a saúde dos seus usuários. / 1,3-dimethylamylamine (DMAA) is a stimulant that started to be added in dietary supplements and weight loss compounds since 2006 and is widely consumed by athletes and the USA army. However, after reports of toxicity DMAA has been banned by regulatory agencies in Brazil and United States. However, even after its prohibition, DMAA still can be found in dietary supplements. Its origin was related to the essential oils of Pelargonium graveolens, and, however, many authors questioned the results and its natural origin. In addition, the essential oils of species of Pelargonium, had their antimicrobial activity reported against bacteria and fungi. Considering the aspects mentioned, this work aimed to determine the presence of DMAA in the essential oils by GC-MS, DART-MS/MS and LC-MS/MS; as well as in the leaves of the same species using the headspace extraction, previously optimized, followed by analysis through GC-MS. It has also been proposed to investigate the antifungal activity of essential oils of P. graveolens from different origins and develop a formulation containing an oil nanoemulsion for the treatment of vaginal candidiasis. Finally, it aimed to develop a screening method to evaluate the presence of DMAA and other stimulants in seized dietary supplements by DART-MS/MS. The results showed that DMAA is not present in the Rio Grande do Sul’s essential oils of Pelargonium spp. obtained by hydrodistillation. After optimization through experimental design, the headspace technique proved to be effective in extracting volatile constituents present in the leaves and, however, DMAA was not detected, as well as in commercial essential oils of P. graveolens from Brazil, China, Egypt, South Africa, Albania and Reunion Islands. The essential oils presented antifungal activity against five Candida species. Furthermore, this antifungal effect presented better results with the nanoformulation containing essential oil. DART-MS/MS screening was effective in detection of DMAA, ephedrine, synephrine, caffeine, sibutramine and methylphenidate in seized dietary supplements, showing positive results for all stimulants. Based on the results obtained and proposed objectives, it was verified that even after using three different analytical techniques and a new alternative for volatile constituents extraction, DMAA was not found in essential oils and leaves of Pelargonium spp., corroborating with other studies carried out, and indicating that its origin is not natural in these species. The final formulation containing the nanoemulsion with the essential oil had antifungal activity superior compared to 14 dispersed essential oil. The analysis of seized samples showed that even after its prohibition by regulatory agencies, supplements containing DMAA and other stimulants are still commercialized, representing a major health risk for their users.