Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-06-21T15:09:18Z No. of bitstreams: 1 rodrigoluizfabri.pdf: 7094274 bytes, checksum: 484f776d7feb93bcc95fb361ef5dc745 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-07T19:10:25Z (GMT) No. of bitstreams: 1 rodrigoluizfabri.pdf: 7094274 bytes, checksum: 484f776d7feb93bcc95fb361ef5dc745 (MD5) / Made available in DSpace on 2017-08-07T19:10:25Z (GMT). No. of bitstreams: 1 rodrigoluizfabri.pdf: 7094274 bytes, checksum: 484f776d7feb93bcc95fb361ef5dc745 (MD5) Previous issue date: 2013-03-01 / FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais / Mitracarpus frigidus (Willd. ex Reem Schult.) K. Schum. (Rubiaceae) é uma espécie nativa, amplamente distribuída por todo território brasileiro, com poucos relatos na literatura sobre seu potencial químico-biológico. Estudos preliminares relataram que a planta apresenta atividade antimicrobiana, leishmanicida e antioxidante, e tem como principais constituintes alcaloides, quinonas, flavonoides, terpenos e esteroides. Com o intuito de dar continuidade aos estudos envolvendo esta espécie, este trabalho teve como objetivos avaliar sua toxidade e seu potencial farmacológico, bem como e isolar e caracterizar suas substâncias bioativas. Do extrato metanólico das partes aéreas de M. frigidus (MFM) foram isolados os triterpenos ácido ursólico e ursolato de metila, e a naftoquinona psicorubrina. Os flavonoides rutina e campferol também foram identificados. Utilizando essas substâncias como marcadores, MFM foi padronizado e avaliado para as atividades esquistossomicida, anti-inflamatória, laxativa e citotóxica. Além disso, sua toxidade aguda (DL50) e subcrônica foram avaliadas. O potencial biológico e a composição química do extrato em hexano e do óleo essencial da espécie também foram analisados. MFM apresentou atividade esquistossomicida tanto in vitro quanto in vivo com considerável redução da carga parasitária. Também foi observada atividade anti-inflamatória aguda e crônica para os modelos testados, sendo a resposta aguda mais expressiva. MFM inibiu o processo de migração celular mediado pela inflamação e também diminuiu o processo oxidativo do organismo, evidenciado pela baixa concentração de MDA, catalase e mieloperoxidase. Em relação à expressão de ciclooxigenases, MFM inibiu COX-2. Atividade citotóxica contra células leucêmicas, HL60 e Jurkat, sem indução de apoptose também foi observada. Além disso, MFM induziu aumento do peristaltismo intestinal e da produção de fezes, o que pode estar relacionado com a presença de antraquinonas identificadas em MFM. A partir do estudo toxicológico agudo e subcrônico de MFM foi possível verificar que a planta tem baixa toxidade (DL50 > 2000 mg/Kg) e que não provocou alterações bioquímicas e hematológicas durante 42 dias de experimento. As substâncias ácido ursólico, ursolato de metila e psicorubrina, isoladas de MFM, apresentaram atividade citotóxica para células tumorais e para diferentes espécies de Leishmania. Da mesma forma, o extrato em hexano apresentou atividade antimicrobiana e leishmanicida, com especificidade e seletividade para formas intracelulares (amastigotas). O óleo essencial apresentou atividade antifúngica e leishmanicida. Linalol e acetato de eugenol foram identificados como as substâncias majoritárias. A partir deste trabalho, foi possível concluir que a espécie M. frigidus, que não possui relatos de uso na medicina popular, pode ser uma fonte alternativa de estudos para o tratamento de diversas doenças, como as provocadas por fungos, bactérias e parasitas, além de patologias relacionadas a processos inflamatórios. / Mitracarpus frigidus (Willd. ex Schult Reem.) K. Schum. (Rubiaceae) is a native species, widely distributed throughout the Brazilian territory, with few reports in the literature on its chemical-biological potential. Previous studies reported that the plant has antimicrobial, antioxidant and leishmanicidal, and that its main constituents are alkaloids, quinones, flavonoids, terpenes and steroids. In order to continue the studies involving this species, this work aimed to evaluate its toxicity and pharmacological potential, as well as to isolate and characterize its bioactive compounds. From the methanolic extract of the aerial parts of M. frigidus (MFM) the triterpenes ursolic acid and methyl ursolate, and the naphthoquinone psycorubrin were isolated. The flavonoids rutin and kaempferol were also identified. Using these compounds as markers, MFM was standardized and evaluated for schistosomicidal, anti-inflammatory, laxative and cytotoxic activity. Moreover, its acute (LD50) and subchronic toxicity were evaluated. The biological potential and chemical composition of the hexane extract and the essential oil of this species were also analyzed. MFM showed both in vitro and in vivo schistosomicidal activity with considerable reduction in parasite burden. Antiinflammatory activity was also observed for the acute and chronic models tested, with a more proeminent acute response. Those results demonstrated that MFM inhibited the inflammation cell mediated migration and also decreased the oxidative process of the organism, as evidenced by the observed low concentration of MDA, catalase and myeloperoxidase. Regarding the expression of cyclooxygenase, MFM inhibited COX 2. Cytotoxic activity against leukemia cells, HL60 and Jurkat, without apoptosis induction was also observed. Furthermore, MFM induced increase of intestinal peristalsis and production of faeces, which can be related to the presence of the anthraquinones identified in MFM. From the acute and subchronic toxicity study of MFM it was possible to verify that the plant has low toxicity (LD50 > 2000 mg/kg) and caused no hematological and biochemical changes during 42 days of experiment. The compounds ursolic acid, methyl ursolate and psycorubrin, isolated from MFM, showed cytotoxicity against different tumor cells and species of Leishmania. Likewise, the hexane extract showed antibacterial and antileishmanial properties, with specificity and selectivity for intracellular forms (amastigotes). The essential oil showed antifungal and leishmanicidal activity. Linalool and eugenol acetate were identified as its major compounds. In conclusion, the species M. frigidus, which has no reports on its use in traditional medicine, could be an alternative source of study for the treatment of various diseases such as those caused by fungi, bacteria and parasites, and pathologies related to inflammatory processes.

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-04-05T18:07:31Z No. of bitstreams: 1 Almeida, Kessiane Belshoff de [Dissertação, 2014].pdf: 1092616 bytes, checksum: ecf1d02bbd15139a6b65760edc3d1bb8 (MD5) / Made available in DSpace on 2017-04-05T18:07:31Z (GMT). No. of bitstreams: 1 Almeida, Kessiane Belshoff de [Dissertação, 2014].pdf: 1092616 bytes, checksum: ecf1d02bbd15139a6b65760edc3d1bb8 (MD5) / A nanoencapsulação de substâncias lipofílicas e sua posterior incorporação em formulações tópicas semissólidas oferece uma alternativa tecnologicamente viável para modular a permeação do ativo, melhorar sua distribuição na superfície da pele, reduzir sua toxicidade e conferir proteção frente a fatores extrínsecos. O polímero poli (ácido lático-co-glicolídeo) (PLGA) tem sido amplamente empregado na preparação de nanossistemas carreadores de fármacos em função de suas pronunciadas características de biocompatibilidade e biodegradabilidade. O presente estudo objetivou desenvolver um hidrogel para incorporação do óleo essencial de Cymbopogon citratus (OECc) encapsulado em nanopartículas poliméricas e avaliar sua atividade frente ao vírus Herpes simplex (HSV) tipos 1 e 2. Inicialmente, procedeu-se a extração de OECc por hidrodestilação, seguindo-se sua caracterização química por Cromatografia Gasosa de Alta Resolução acoplada ao Espectrômetro de Massas (CGAR-EM). Nanopartículas contendo OECc (NPOE) e nanopartículas branco (NP) foram preparadas pela técnica de emulsificação-difusão do solvente empregando PLGA, como polímero, e álcool polivinílico (PVA) como estabilizante. A distribuição de tamanho e o potencial zeta das partículas foram avaliados, respectivamente, pelas técnicas de espalhamento de luz dinâmico e mobilidade eletroforética, e sua eficiência de encapsulação determinada após extração com solvente, empregando metodologia analítica desenvolvida por espectrofotometria UV-Vis. Após preparo e caracterização, NPOE foram incorporadas a gel hidrofílico de Carbopol® Ultrez 10 NF (HNPOE), a fim de investigar a estabilidade da formulação nanoestruturada frente ao armazenamento e o perfil de liberação in vitro do óleo a partir da nanopartícula. Adicionalmente, hidrogel base (HB), hidrogel contendo o óleo livre (HOE) e o mesmo contendo nanopartículas branco (HNP) foram desenvolvidos como controles. A atividade inibitória das formulações frente HSV-1 e -2 sensíveis ao aciclovir foi avaliada por redução do título viral em placa, utilizando-se células Vero como sistema hospedeiro, após prévia determinação da citotoxicidade das amostras. O rendimento do processo extrativo foi de 0,37% e a análise qualitativa do óleo por CGAR-EM exibiu o citral como seu constituinte químico majoritário, correspondendo a 89,57 % da área relativa do cromatograma. Partículas em escala nanométrica e potencial zeta negativo foram obtidas, com um conteúdo de OECc de 58,59 ± 0,85 mg/g e eficiência de encapsulação de 28,48% ± 0,40%. Os hidrogéis desenvolvidos exibiram características apropriadas para aplicação cutânea, as quais se mantiveram inalteradas durante os 60 dias de armazenamento a 4 °C. Cabe salientar que as formulações HNPOE e HOE apresentaram redução significativa (p<0,05, t-Student) no conteúdo de óleo volátil em relação ao teor inicial, permanecendo subsequentemente constante durante o estudo de estabilidade. A análise do perfil de liberação in vitro de OECc a partir das formulações demonstrou padrão bifásico, com burst inicial e posterior fase de liberação sustentada, onde NPOE seguiu modelo cinético de Hixon-Crowell, e HNPOE e HOE modelo de Higuchi. Além disso, as mesmas exibiram comportamento anômalo, dependente dos mecanismos de difusão e erosão polimérica. Na avaliação da atividade antiviral, HNPOE foi capaz de inibir mais eficientemente ambas as estirpes virais em concentração não citotóxica de óleo, inferior às empregadas nas demais formulações. Estes resultados evidenciam o potencial do nanogel em proteger, modular a liberação e otimizar a atividade do óleo essencial frente ao vírus Herpes simplex / The nanoencapsulation of lipophilic substances and their subsequent incorporation into semisolid topical formulations offers a technologically feasible alternative to modulate the permeation of active, improve its distribution on the surface of the skin, reduce their toxicity and confer protection against extrinsic factors. The polymer poly (lactic acid-co-glycolide) (PLGA) has been widely employed in the preparation of drug carriers nanosystems due to their pronounced biocompatibility and biodegradability. The present study aimed to develop semisolid hydrogel incorporating of Cymbopogon citratus essential oil (CcEO) associated with polymeric nanoparticles and assess their activity against Herpes simplex virus (HSV) types 1 and 2. Initially, it was proceeded to extract the CcEO by hydrodistillation, following their chemical characterization by High-Resolution Gas Chromatography coupled to Mass Spectrometer (HRGC-MS). Nanoparticles containing CcEO (NPEO) and white nanoparticles (NP) were prepared by emulsification-diffusion of the solvent using PLGA as polymer and polyvinyl alcohol (PVA) as a stabilizer. The size distribution and zeta potential of the particles were determined, respectively, by dynamic light scattering and electrophoretic mobility, and its encapsulation efficiency was given by solvent extraction technique, using analytical methodology developed by UV-Vis spectrophotometry. After preparation and characterization, NPEO were incorporated into a hydrophilic gel Carbopol® Ultrez 10 NF (HNPEO) in order to investigate the stability of the nanostructured formulation across the storage and in vitro release profile of the oil from the nanoparticle. In addition, hydrogel base (HB), hydrogel containing free oil (HEO) and the same unloaded nanoparticles (HNP) were developed as controls. The inhibitory activity of the formulations against HSV-1 and -2 sensitive to acyclovir was assessed by viral titer reduction using Vero cells as a host system after prior determination of the cytotoxicity of the samples. The yield of the extraction process was 0.37% and the qualitative analysis of the oil by HRGC-MS showed the citral as its major chemical constituent, accounting for 89.57% of the chromatogram relative area. Particles on the nanometer scale and negative zeta potential were obtained with CcEO content of 58.59 ± 0.85 mg/g and encapsulation efficiency of 28.48 ± 0.40%. The developed hydrogels exhibit suitable characteristics for cutaneous application, which remained unchanged during the 60 days of storage at 4°C. Is worth emphasizing that the HNPEO and HEO formulations showed a significant reduction (p<0.05, Student's t test) in the content of volatile oil in relation to the initial rate, subsequently remaining constant during the stability study. The analysis of the in vitro release profile CcEO from the formulations showed a biphasic pattern with an initial burst and subsequent sustained release phase, which followed NPEO kinetic model Hixon-Crowell, and HNPOE and HOE Higuchi model. Moreover, they exhibited anomalous behavior, dependent on the mechanisms of diffusion and polymer erosion. In the evaluation of antiviral activity, HNPEO was able to more efficiently inhibit both viral strains in the non-cytotoxic oil concentration lower than used in the other formulations. These results highlight the potential of nanogel in protecting, modulating the release profile and improve the activity of the essential oil against Herpes simplex virus

Made available in DSpace on 2014-08-20T14:38:04Z (GMT). No. of bitstreams: 1 tese_eduardo_negri_mueller.pdf: 1762619 bytes, checksum: a9e8ea647471ee1e308c249dfa91f25d (MD5) Previous issue date: 2011-11-16 / The aim of this study was to evaluate the microenvironment of the ear canal of dogs and determine the effect of Rosmarinus officinalis L. (rosemary) and Triticum vulgare (wheat) in healthy ears and in the treatment of experimental noninfectious external otitis of Wistar rats. For analysis of the microenvironment of the ear canal 141 healthy dogs were used, 30 to evaluate the ear canal pH and 111 to measure the ear temperature. Dog ears with absent to mild, moderate or intense cerumen were studied, and the auditory canal was washed for pH measurements. The rectal temperature (TR) was measured with a mercury thermometer and the ear temperature with an infrared thermometer. The difference between TR and TAM (average ear temperature), was classified in the following scores: A=TAM<TR until 1.2°C and B=TAM<TR more than 1.2°C, and these results were related to ear type. For the evaluation of the effects of plant extracts in the experimental external otitis in Wistar rats an inoculation of 80μL of croton oil 5% in acetone was performed and in 24 hours the clinical score of the ears was evaluated. The animals were divided in 5 groups and the topic treatment was realized for up to seven days with rosemary aqueous extract 25% in propylene glycol (Group I), rosemary essential oil 25% (Group II), wheat aqueous extract 0.2% in propylene glycol (Group III), propylene glycol (Group IV) and NaCl 0.9% (Group V). At days four (4d), six (6d) and ten (10d), the ears of four animals were clinically evaluated and the animals were euthanized for hystopathological evaluation. For the evaluation of the effects of plant extracts on the healthy ear 48 Wistar rats were distributed in five groups according previously described and classified for clinical score. In the analysis of the microenvironment of the ear canal the pH values ranged from 5.0 to 8.0 and there was a negative correlation between cerumen quantity and pH (p=0.0271). The ear type did not affect the TAM and predominated ears score B regardless of the type. In the evaluation of the treatment for experimental otitis groups I, II and V had an increase in clinical score until the fourth day then decreased gradually. Groups III and IV had the scores lower tha day 0 for almost all experimental period. Group III showed a progressive decrease; group II showed the highest score at days four and six and the group V at day ten. There was hyperplasia in all groups at the end of the experiment, and in groups II and IV it was more frequent. All groups had otitis with predominantly histiocytic infiltrate at the beginning, although at 10 days with a suppurative infiltration in groups II, III and V. In healthy ears at four days, all groups showed an increase of clinical scores, which was higher in groups I (2.6), II (3.6) and III (2.4), but only the essential oil of rosemary 25% in propylene glycol was topically irritant. There was a progressive decrease of the scores in groups I, II and III starting at day four and at day 10 the highest score was observed in groups I (2.0) and II (2.0). It is concluded that the pH of the ear canal of dogs is neutral and can acidify with increased amount of cerumen; the average ear temperature is lower than the rectal temperature by 1.53°C±0.75 and the conformation of the ear does not influence the average ear temperature; aqueous extract of Rosmarinus officinalis L. 25% and aqueous extract of Triticum vulgare 0.2% reduced the clinical scores, while the essential oil of Rosmarinus officinalis L. 25% is not effective in noninfectious otitis externa in Wistar rats and the aqueous extract of Rosmarinus officinalis L. 25% and the aqueous extract of Triticum vulgare 0.2% are non-irritating topics, while the essential oil of Rosmarinus officinalis L. 25% irritates the auditory epithelium. / Objetivou-se avaliar o microclima do canal auditivo de cães e determinar o efeito do Rosmarinus officinalis L. (alecrim) e do Triticum vulgare (trigo) em orelhas hígidas e no tratamento da otite externa experimental não infecciosa de ratos Wistar. Para análise do microclima do canal auditivo foram considerados 141 cães hígidos, destes 30 utilizados para avaliação do pH do canal auditivo e 111 para aferição das temperaturas auditivas. Os cães foram distribuídos em dois grupos de acordo com a quantidade de cerúmem ausente a leve e moderado a intenso e o canal auditivo foi lavado para aferição do pH. A diferença entre a temperatura retal (TR), aferida com termômetro coluna de mercúrio e a temperatura auditiva média (TAM), aferida com termômetro infravermelho, foi avaliada para classificação em escores: A=TAM<TR em até 1,2ºC e B=TAM<TR em mais de 1,2ºC, relacionando-as com a conformação da orelha. Para avaliação do efeito dos extratos vegetais na otite externa, 64 ratos Wistar foram submetidos à inoculação no conduto auditivo de 80µL de óleo de cróton 5% em acetona e, em 24h as orelhas foram avaliadas por escore clínico. Os animais foram divididos em cinco grupos, sendo o tratamento tópico realizado por até sete dias com extrato aquoso de alecrim 25% em propilenoglicol (grupo I), óleo essencial de alecrim 25% em propilenoglicol (grupo II), extrato aquoso de trigo 0,2% em propilenoglicol (grupo III), propilenoglicol (grupo IV) e com NaCl 0,9% (grupo V). Aos quatro (4d), seis (6d) e dez dias (10d) as orelhas de quatro animais de cada grupo foram avaliadas clinicamente e, os mesmos foram eutanasiados para histopatologia. A avaliação do efeito dos extratos vegetais na orelha hígida foi realizada utilizando 48 ratos Wistar, os quais foram distribuídos em cinco grupos de acordo com o tratamento, conforme descrito acima e classificados em escore clínico. Na análise do microclima do canal auditivo observou-se que o pH variou de 5,0 a 8,0 e houve correlação negativa entre quantidade de cerúmen e pH (p=0,0271). Ainda, o tipo de orelha não influenciou a TAM e predominaram orelhas no escore B independente do tipo. Na avaliação do tratatmento da otite externa experimental os grupos I, II e V aumentaram os escores aos quatro dias, em seguida decresceram. Os grupos III e IV mantiveram os escores menores que o dia zero por quase todo o período experimental. O grupo III apresentou diminuição progressiva, o II apresentou o escore mais elevado aos quarto e seis dias e o grupo V aos 10 dias. Havia hiperplasia em todos os grupos e no final do experimento, nos grupos II e IV em maior freqüência. Todos os grupos tinham otite no início com predomínio de infiltrado histiocitário, embora aos 10 dias com infiltrado supurativo nos grupos II, III e V. Nas orelhas hígidas aos quatro dias, todos os grupos apresentaram aumento dos escores clínicos sendo maior nos grupos I(2,6), II(3,6) e III(2,4), porém somente o óleo essencial de alecrim 25% em propilenoglicol apresentou efeito irritante tópico. Houve uma diminuição progressiva dos escores nos grupos I, II e III a partir do dia quatro e no dia 10 o maior escore foi observado nos grupos I (2,0) e II (2,0). Concluise que o pH do canal auditivo de cães é neutro e pode acidificar com o aumeno da quantidade de cerúmen; a temperatura auditiva média é menor que a retal 1,53°C±0,75 e a conformação da orelha não influencia na temperatura auditiva média em cães; o extrato aquoso de Rosmarinus oficinalis L. 25% e o extrato aquoso de Triticum vulgare 0,2% reduzem os escores clínicos, enquanto o óleo essencial do Rosmarinus oficinalis L. 25% não é efetivo na otite externa não infecciosa em ratos Wistar e, o extrato aquoso de R. officinalis L. 25% e o extrato aquoso de T. vulgare 0,2% não são irritantes tópicos, enquanto que o óleo essencial de R. officinalis L. 25% irrita o epitélio auditivo.

Made available in DSpace on 2014-08-20T13:59:06Z (GMT). No. of bitstreams: 1 tese_milene_conceicao_lima.pdf: 1117046 bytes, checksum: 61412e1bba4614df5d623d5245902770 (MD5) Previous issue date: 2013-04-05 / Several abiotic factors can coordinate or alter metabolic processes of growth and development plants such light, temperature, water availability, climate, soil nutrition and inorganic compounds. The objective of this study was to evaluate performance of A. millefolium L. (yarrow) against abiotic factors. To achieve this objective were have conducted two experiments in which the first was observed antimicrobial activity in vitro and essential oil yield of yarrow grown with nitrogen levels (0, 75 and 150 kg urea ha-1) and subjected different levels of shading 0% (full light) and 70% (70% shading). The second experiment were observed growth characteristics, production photosynthetic pigments, antioxidant enzymes activity, concentrations hydrogen peroxide, malondialdehyde, ascorbate and deidroascorbato, antioxidant activity by method DPPH and secondary metabolites synthesis in yarrow cultivated under light conditions (full light and blue, red and black shade net) and subjected different collection periods (eight weeks shading, two and four weeks after shading). The results show that oil yield extracted from flowers was stimulated by medium nitrogen level and full light increased the oil yield extracted from leaves. The highest essential oil yield was observed in reproductive structures. Antimicrobial activity of oil was confirmed against microorganisms species. Yeast strains were inhibited by the lowest concentration. The dry mass of leaves and flowers was favored in plants grown under full light. The content of chlorophyll a and b were higher in plants cultivated under blue net, total chlorophyll content was lower in plants under full light and plants two weeks after shading was increase of chlorophyll a, b and total, equivalent did not shaded plants. Catalase activity increased plants acclimated to high light intensity and low light did not stimulate ascorbate peroxidase, but increased superoxide dismutase and content of hydrogen peroxide. Lipid peroxidation was reduced presence of the spectrum blue and red light and low light intensity generated by nets black and blue, reduced accumulation of ascorbate. Content of deidroascorbato increased acclimation process to sunlight and antioxidant activity was more pronounced extracts of flowers. Levels total phenols and flavonoids were stimulated by full light. Oil yield extracted from flowers was higher than leaves. Major metabolite oil extracted from leaves was farnesol and flowers were farnesol and chamazulene. Results for medicinal species A. millefolium L. indicated that light intensity, light quality, collection periods and nitrogen fertilization influence growth, photosynthetic pigments production, secondary metabolites and antioxidant response. / Diversos fatores abióticos podem coordenar ou alterar os processos metabólicos de crescimento e desenvolvimento em plantas como a luz, temperatura, disponibilidade hídrica, clima, nutrição do solo e compostos inorgânicos. O objetivo deste estudo foi avaliar o comportamento de A. millefolium L. (mil-folhas) frente aos fatores abióticos. Para atingir este objetivo foram realizados dois experimentos, na qual no primeiro foi verificada a atividade antimicrobiana in vitro e rendimento do óleo essencial de mil-folhas cultivada com concentrações de nitrogênio (0, 75 e 150 Kg ureia ha-1) e submetida a diferentes níveis de sombreamento 0% (luz plena) e 70% (sombreamento 70%). No segundo experimento foram avaliadas características de crescimento, produção de pigmentos fotossintéticos, atividade de enzimas antioxidantes, teores de peróxido de hidrogênio, malondialdeído, ascorbato e deidroascorbato, atividade antioxidante pelo método DPPH e síntese de metabólitos secundários em mil-folhas cultivada sob condições de luminosidade (luz plena e sombreamento com malha azul, vermelha e preta) e submetida a diferentes períodos de coleta (oito semanas de sombreamento, duas e quatro semanas após o sombreamento). Os principais resultados mostram que o rendimento do óleo extraído de flores foi estimulado pela concentração nitrogenada intermediária e o extraído de folhas foi favorecido pela luz plena, sendo maior nas estruturas reprodutivas. A atividade antimicrobiana do óleo foi confirmada frente aos microrganismos, sendo as cepas fúngicas inibidas pela menor concentração. A massa seca das folhas e flores foi favorecida nas plantas cultivadas com luz plena. Os teores de clorofila a e b foram maiores nas plantas cultivadas com a malha azul, o conteúdo de clorofila total foi menor nas plantas sob luz plena e em plantas com duas semanas após o sombreamento ocorreu um incremento de clorofila a, b e total, equivalendo-se às plantas não sombreadas. A atividade da catalase aumentou em plantas aclimatadas em alta intensidade de luz e a baixa luminosidade não estimulou a ascorbato peroxidase, mas incrementou a superóxido dismutase e o teor de peróxido de hidrogênio. A peroxidação lipídica foi reduzida na presença dos espectros de luz azul e vermelha e a baixa intensidade de luz, gerada pelas malhas preta e azul, reduziu o acúmulo de ascorbato. O conteúdo de deidroascorbato aumentou em processo de aclimatação à luz solar e a atividade antioxidante foi mais acentuada em extratos de flores. Os teores de fenois e flavonoides totais foram estimulados pela luz plena. O rendimento do óleo extraído de flores foi maior em relação ao de folhas. O metabólito majoritário do óleo extraído de folhas foi o farnesol e os extraídos de flores foram o farnesol e camazuleno. Os resultados encontrados para a espécie medicinal A. millefolium L. indicaram que a intensidade luminosa, qualidade de luz, períodos de coleta e fertilização nitrogenada influenciam no crescimento, produção de pigmentos fotossintéticos, de metabólitos secundários e resposta antioxidante.

The Myrtaceae family has 145 genera. In Brazil are recorded 23 genera and about 1,013 species. Several plants of this family are used in food stuffs, for example, Psidium guajava L. fruits, commonly known as guava. In folk medicine Psidium guajava L. leaves are used for colic, colitis, diarrhea, dysentery, and stomach illness. The objective of this study was to evaluate the biological effects of essential oil (OEPGP) and hydro alcoholic extract (HEPG) of Psidium guajava var. pomifera L. To evaluate the OEPGP bioinseticide effect Drosophila melanogaster were used. The flies exposure to OEPGP was performed by a fumigation protocol: Adult flies (males and females) were placed in glass vials containing a filter paper soaked in 1% sucrose. Then, a filter paper was set on the inner side of the cover containing the OEPGP (0, 3, 7.5, 15, 23.5 and 30 μg/ml). The flies mortality was assessed 6, 12, 24 and 48 h. In turn, to assess antioxidant and chelating action against inorganic mercury, DPPH, TBARS, and 1,5 diphenylcarbazone assays were carried out using 0, 0.1, 0.2, and 0.4 mM HgCl2 and 0, 25, 33, 50, 100 and 200 μg/ml of EHAPGP. Cell growth was assessed using Sacharomices cerevisiae, which were exposed to concentrations of 100, 125 and 150 mM of HgCl2 and 0, 0.25, 0.5, 1, 1.25, 2.5, and 5 μg/ml of HEPG. Our results showed that OEPGP has a bioinseticide action when flies were exposed to concentrations of 23.5 and 30 μg/ml. Locomotor activity was impaired at concentrations of 15, 23.5, and 30 μg/ml during the first 6 hours of treatment. The OEPGP caused an increase in TBARS levels and in antioxidant enzymes activities. In turn, the HEPG has high concentrations of phenols and flavonoids and showed antioxidant activity, as demonstrated in the DPPH and TBARS assays. The chelating action of HEPG was proven by the test of 1.5 difenilcarbazone and the dosages of Hg levels by ICP-AES. The HEPG partially prevented the cell death caused by HgCl2. Our study indicated that HEPG has antioxidant and HgCl2 protection purposes, while the essential oil presents a bioinseticide action to trigger a pro-oxidant action. / A família Myrtaceae possui 145 gêneros. No Brasil são registrados 23 gêneros e cerca de 1.013 espécies. Muitas plantas desta família são utilizadas na alimentação, como os frutos da Psidium guajava L., popularmente conhecidos como goiaba. As folhas da goiabeira são utilizadas na medicina popular para cólicas, colite, diarréia, disenteria e dor de barriga. O objetivo deste trabalho foi avaliar os efeitos biológicos do óleo essencial (OEPGP) e do extrato hidro alcóolico (HEPG) das folhas de Psidium guajava var. pomifera L. Para avaliar a ação bioinseticida do OEPGP foram utilizadas Drosophila melanogaster. A exposição das moscas ao OEPGP foi realizada por um protocolo de fumigação: moscas adultas (machos e fêmeas) foram colocadas em frascos de vidro, contendo um papel filtro embebido em 1% de sacarose. Um papel filtro foi fixado no lado interior da tampa para aplicação de diferentes doses de óleo essencial (0, 3, 7,5, 15, 23,5 e 30 μg/mL). A mortalidade das moscas foi avaliada 6, 12, 24 e 48 h. Por sua vez, para avaliar a ação antioxidante e quelante contra o mercúrio foram elaborados ensaios de DPPH, TBARS e 1,5-difenilcarbazona, nas concentrações de 0, 0,1, 0,2 e 0,4 mM de HgCl2 e concentrações de 0, 25, 33, 50, 100 e 200 μg/mL do HEPG. O crescimento celular foi avaliado utilizando Sacharomices cerevisae, os quais foram expostos a concentrações de 100, 125 e 150 μM de HgCl2 e 0, 0,25, 0,5, 1, 1,25, 2,25 e 5 μg/mL de HEPG. Os resultados mostram que o OEPGP apresenta uma ação bioinseticida quando as moscas foram expostas às concentrações de 23,5 e 30 μg/mL. A atividade locomotora foi comprometida nas concentrações de 15, 23,5, e 30 μg/mL nas primeiras 6 h de tratamento. O OEPGP causou aumento nos níveis de TBARS e na atividade das enzimas antioxidantes GST e CAT. Por sua vez, o HEPG apresenta elevadas concentrações de fenóis e flavonoides e apresentou atividade antioxidante, comprovada nos ensaios de TBARS e DPPH. A ação quelante do HEPG foi comprovada pelo ensaio da 1,5-difenilcarbazona e pelas dosagens dos níveis de Hg por ICP-AES. O HEPG evitou parcialmente a morte celular causada por HgCl2. Nosso estudo indicou que HEPG tem efeitos antioxidantes e de proteção contra o estresse oxidativo causado pelo HgCl2, enquanto que o óleo essencial apresenta uma ação bioinseticida por desencadear uma ação pró-oxidante.

En Afrique sud- saharienne, plusieurs méthodes de protection des stocks alimentaires sont utilisées ; parmi elles, les insecticides synthétiques qui représentent un risque réel pour la santé humaine. Pour apporter une alternative à ces insecticides chimiques potentiellement dangereux, le présent travail a pour objectif de développer des bio-insecticides en formulant des poudres par adsorption d’huiles essentielles sur des argiles. Deux argiles naturelles du Cameroun ont été utilisées comme adsorbants des composés terpéniques des huiles essentielles de Xylopia aethiopica et Ocimum gratissimum pour la préparation d’insecticides. Dans le but d’améliorer leur capacité d’adsorption, ces argiles ont été traitées par la soude et l’acide sulfurique. Elles ont également été modifiées par des solutions de polycations d’aluminium ou de fer de rapport molaire varié ainsi que par des cations d’alkylammoniums. Une bentonite commerciale a été utilisée comme référence dans le suivi des modifications. Ces différentes matrices ont été caractérisées avant et après modifications par diffraction des rayons X, adsorption-désorption d’azote, spectroscopie infra rouge à transformée de Fourier, analyses thermogravimétriques et différentielles, et photométrie de flamme. Il ressort de la caractérisation que l’échantillon de Wak est majoritairement constitué de kaolinite, et celui de Maroua de montmorillonite. Le traitement par l’acide sulfurique et par les polycations métalliques entraînent une augmentation de la surface spécifique des argiles, tandis que la soude et les cations d’alkylammoniums la réduisent fortement. La surface spécifique de la montmorillonite passe de 82 m².g-1 à 4,5 m².g-1 après traitement au céthyl triméthyl ammonium (CTMA). Les diffractogrammes montrent une augmentation de la distance interfoliaire des argiles étudiées après modification aux cations alkylammoniums. La distance interfoliaire augmente de 5,5 Å et de 10,6 Å respectivement pour la montmorillonite et la bentonite traitée par le CTMA. Les argiles de type smectite après modifications aux polycations métalliques, présentent en revanche un étalement du pic caractéristique de l’espace interfoliaire. Les cations alkylammoniums entraînent donc une intercalation effective des molécules de CTMA et phényl triméthyl ammonium (PTMA) entre les feuillets de montmorillonite et de bentonite tandis que les polycations métalliques entraînent une exfoliation du matériau argileux. Les argiles-alkylammoniums présentent les plus grandes capacités d’adsorption des composés terpéniques malgré les plus faibles surfaces spécifiques. Ainsi, face aux composés terpéniques, la capacité d’adsorption des argiles ne dépend pas uniquement de la surface spécifique mais également de l’espacement interfoliaire de l’adsorbant ainsi que de l’affinité des molécules d’adsorbât vis-à-vis de l’adsorbant. Des tests insecticides ont montré que les formulations à base de la Mont-CTMA présentent une toxicité plus stable que celle préparée à partir de l’argile brute (Mont-Na). La formulation Mont-Na-HE perd la totalité de son activité insecticide au bout de 30 jours de conservation dans les boîtes ouvertes. La formulation Mont-CTMA-HE par contre n’en perd qu’environ 60% dans les mêmes conditions. La rémanence de la formulation varie avec l’adsorbant utilisé ; car l’effet insecticide de l’huile essentielle d’O. gratissimum persiste pendant 107 jours lorsqu’elle est fixée sur la Mont-CTMA, tandis que fixée sur l’argile brute, elle perd son activité au bout de 45 jours. Ces résultats nous permettent d’affirmer que les argiles modifiées augmentent la durée de l’effet insecticide des huiles essentielles et peuvent être utilisées pour une application industrielle dans la production des bio-insecticides. / In sub-Saharan Africa, several methods to protect food stocks are used ; amongst them are synthetic insecticides. These synthetic insecticides pose high real risk to human health. This study aims at providing an alternative to the dangerous synthetic chemical insecticides, by developing bioinsecticides formulating powders through adsorption of essential oils on Cameroonian clays. Two natural clays collected in Cameroon were used as adsorbents of terpene compounds of Xylopia aethiopica and Ocimum gratissimum essential oils, for the preparation of insecticides. The clay fractions were obtained from soil aggregates by sedimentation. In order to improve their adsorption capacity, these clays were treated with sodium hydroxide and sulfuric acid. They were also modified with solutions of aluminum, iron polycations with molar ratio varied and alkylammoniums cations. A commercial bentonite was used as reference for these modifications. These different adsorbents materials were characterized before and after modifications by X-ray diffraction, nitrogen adsorption-desorption, infrared Fourier transform spectroscopy, thermal analysis, and flame photometry. According to the characterizations of the clay materials, Wak sample contained more kaolinite clay, while Maroua sample contained more montmorillonite clay. The sulfuric acid and metallic polycations caused an increase in the specific surface of clays, while sodium hydroxyl and alkylammoniums cations greatly reduced the SBET. For example, the Mont-Na+ SBET decreased from 82 m². g-1 to 4.5 m².g-1 after treatment with cetyl trimethyl ammonium (CTMA). Diffractogrammes showed spacing of smectites interlayer (montmorillonite and bentonite) after treatments with alkylammoniums cations at 5.5 Å and 10.6 Å respectively, for the interlayer space of Mont- CTMA and Bentonite- CTMA clays. After treatments with metallics polycations, smectite clays show a broadening of characteristic peak. Alkylammoniums cations led to an effective insertion of CTMA molecules and phenyl trimethyl ammonium (PTMA) between the interfoliar space of montmorillonite and bentonite while the metal polycations led to an exfoliation of the same clays. The interreticular spacings of kaolinite remain unchanged after any treatments. The kinetic model of pseudo second order is applicable to the adsorption of terpenic compounds by clays. Finding also showed that spread is not the only limiting factor in the process of adsorption of terpene compounds. Alkylammoniums-clays have the largest adsorption capacities of terpene compounds despite the lower specific surface they present. All this allows us to maintain that in the presence of terpenic compounds, the adsorption capacity of clays depends not only on the SBET but more on the interlayer spacing of the adsorbent and on the affinity of the adsorbate molecules toward the adsorbent. Concerning bioassays, it appears that the formulations made with Mont- CTMA presented a more stable toxicity than Mont-Na. Mont-Na-EO formulation loses all its insecticidal activity after 30 days of storage in the open boxes ; while Mont-CTMA- EO formulation only loses about 60% under the same conditions. Moreover, the Mont-Na-EO loses 50% of its insecticidal power (LD50) after 6 days ; while Mont-CTMA- EO loses the same quantity after 16 days when stored in open boxes. The remnance effect of the formulations based on essential oil varied with the adsorbent used. Because the insecticidal effect of O. gratissimum essential oil persisted during 107 days when it is adsorbed on Mont-CTMA, when adsorbed on Mont-Na, it loses all its activity for about 45 days. These results allow us to assert that modified clays increase the duration of the insecticidal effect of essential oils and can be used for industrial application in the production of bio-insecticides based on essential oils.

Les huiles essentielles (HE) de lavande et de lavandin appartiennent au patrimoine de la région méditerranéenne. Comme tout produit naturel ou synthétique à valeur ajoutée, ces HE doivent être contrôlées pour justifier la qualité du produit. Ce contrôle qualité nécessite la mise en place d'une méthodologie analytique fiable. Dans cette étude, une nouvelle approche utilisant les techniques spectroscopiques et chromatographiques et le traitement de données à l'aide d'outils chimiométriques permet de différencier les HE de lavande et de lavandin par origine variétale. Cette discrimination s'effectue grâce à leur empreinte spectrale ou chromatographique. L’intérêt d’utiliser la chromatographie chirale associée à la détection polarimétrique et de la signature chiroptique acquise a également été étudié. Cette étude nous a permis d’identifier des marqueurs métabolomiques qui sont primordiaux pour caractériser les variétés. Les résultats montrent qu'il est possible de discriminer les HE de lavande et de lavandin selon leurs variétés avec une bonne justesse sur l'ensemble des techniques utilisées. / Lavender and lavandin essential oils (EOs) belong to the heritage of the Mediterranean region. Like any natural or synthetic product with an added value, these EOs must be controlled to justify the quality of the product. This quality control requires the establishment of a reliable analytical methodology. In this study, a new approach using spectroscopic and chromatographic techniques for data processing associated to chemometric tools allows to discriminate lavender and lavandin EOs. This discrimination is carried out thanks to their spectroscopic or chromatographic fingerprints. The interest to use the chiral chromatography combined with polarimetric detection and of acquired chiroptical signature was also studied. This methodology has allowed us to identify metabolomic markers which are paramount to characterize the varieties. The results show that it is possible to discriminate the lavender and lavandin EOs according to their varieties with good accuracy on all of the techniques used.

The genus Lippia Houst. (family Verbenaceae Juss.) has been used medicinally for centuries in various cultures. Numerous traditional medicinal applications of the South African Lippias are known, and among these is Lippia scaberrima Sond., the "Beukesbossie". Products recovered from L. scaberrima include essential oils by means of batch microwave distillation and a herbal tea manufactured by various methods. Microwave heating occurs through direct absorbtion of input energy. The whole sample is heated simultaneously with a heating rate much faster than for water distillation. Microwave distillation times can be expected to be less than corresponding water distillation times. A large number of parameters influence essential oil composition and yield. Various geographical, climatic, botanical, production and other parameters were identified and listed. The natural distribution of airdried leaves and flower heads and water as solvent was used in a domestic type microwave oven. Microwave distillation production parameters were studied and optima determined with respect to maximum oil yield. The batch microwave distillation production parameters studied (irradiation time, ratio of water used to plant material used, plant material moisture content, solvent recycling, physical size of plant material, addition of a surface tension modifier, soaking as pretreatment, load size, distillation kinetics and alternative solution medium namely ethanol) lead to a better understanding of microwave essential oil production. It enabled design of prototype continuous microwave distillation apparatus. Mobile, continuous microwave distillation equipment enables extraction of essential oils in a rural environment. It allows rapid investigation of a large variety of plants, and retrieval of essential oils in an economical and uncomplicated manner. Water distillation represents a classical technique for essential oil retrieval whereas microwave distillation is probably the most modern method. Physical and chemical properties of microwave and water distilled oils were determined. This enabled comparison of the microwave technique to a standard, industrially accepted production method. Tea was manufactured from L. scaberrima by means of the green tea, Rooibos tea and black tea methods as a first iteration in the production of a commercial tea. Drying curves was developed as well as appropriate regions of production variables like withering- and fermentation temperatures and times. / Dissertation (PhD(Ing.Chem))--University of Pretoria, 2002. / Chemical Engineering / unrestricted

Les apiaceae peuvent être définies en tant qu' Aroma-Tincto-Oleo-Crop (ATOC), plantes qui contiennent à la fois une huile végétale et une huile essentielle. Appliquer le concept d'agroraffinage aux ATOC revient à proposer un procédé séquentiel alliant une co-extraction huile végétale et huile essentielle à une valorisation des résidus en tant que source de molécules biosourcées et de susbtrat pour la formulation d'agromatériaux . Les objectifs de cette thèse seront donc d'étudier la faisabilité biologique et technologique d'application du concept d'ATOC-raffinage à la coriandre (Coriandrum sativum L.). Le chapitre I présente l'état de l'art bibliographique sur l'extraction et l'analyse des huiles végétales et huiles essentielles de coriandre tandis que dans le chapitre II sont décrits les matériels et méthodes mis en oeuvre au cours de la thèse tant au niveau échantillonnage, extraction, analyse que traitement des données. Le chapitre III est centré sur l'étude des différents paramètres biologiques pouvant influencer la bioaccumulation des huiles végétales et huiles essentielles dans la coriandre (différentes variétés, différents organes de la plante, différents stades de développement biologique) et leur impact sur l'activité anti-oxydante des extraits obtenus à partir des résidus d'extraction. Dans le chapitre IV, la technologie d'extrusion (mono-vis et bi-vis) a été appliquée aux graines de coriandre dans le but d’évaluer la faisabilité du pressage mécanique du fruit de la coriandre pour l’extraction d'une huile végétale aromatisée. L’influence des conditions expérimentales sur le rendement d'extraction en huile végétale (diamètre de buse et distance buse-vis (extrudeur mono-vis) ou configuration de vis, coefficient de remplissage et température de pressage (extrudeur bi-vis)) a été étudiée tandis que la faisabilité de la valorisation du résidu solide d'extraction en agromatériaux (thermo-pressage) a été montrée. / Apiaceae could be defined as Aroma Tincto Oleo Crops (ATOC), e.g. plants containing both vegetable oil and essential oil. Applying agroreffinery concept to ATOC led to propose a sequential fractionation process coupling co-extraction of vegetal oil and essential oil to a valorization of by-product residues as biosourced active molecules and substrates for designing agromaterials. The aim of this thesis is to determine the biological and technological feasability of application of the ATOC-refinery concept to coriander (Coriandrum sativum L.) Chapter I reports a bibliographic state of the art study on extraction and characterisation of coriander vegetal oil and essential oil while chapter II describes materials and methods setting up during the thesis for sampling, extraction, analysis and data processing. Chapter III focus on the study of major various biological parameters influencing bioaccumulation of vegetal oil and essential oil in coriander (different plant varieties, different plant organs, different biological stages) and their impact on anti-oxidant activity of extracts obtained from extraction residues. In chapter IV, coriander fruits are processed by extrusion technology (mono screw and twin-screw extruder) in order to evaluate the feasability of mechanical pressing for extracting a flavored vegetal oil. Influence of operating parameters on vegetal oil extraction yields (nozzle diameter and nozzle/screw distance (single-screw extruder) or screw configuration, device’s filling coefficient and pressing temperature (twin-screw extruder)) is studied while the feasability of valorization of extraction cake as agromaterial (thermopressing) was stated.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / A lot of vegetable species synthesize and accumulate organic substances, such as essential oils, that can be turned in alternative income for rural producers. Pelargonium graveolens L. is a native aromatic species from South Africa, commonly known as geranium. Its essential oil is largely used in perfume, cosmetics and fragrances industries, besides the aromatherapy. The conventional vegetative propagation methods of geranium do not present considerable success, which difficult the production of transplants in large scale. This way it is important to look for alternative production techniques, such as micropropagation, for this commercially important species. The aims of this work were to establish a protocol for micropopagation and acclimatization of geranium, besides investigating of essential oil content and chemical composition. The completely randomized design was used. For the micropropagation assays were tested different concentrations and immersion times of sodium hypochlorite and mercury chloride, different types of explants and MS salts concentrations, besides testing different concentrations of the plant growth regulators BAP, NAA, kinetin and IAA at presence and absence of light. For the acclimatization assay different substrates were tested. To analyze the essential oil we use plants propagated by rooting of cuttings and by micropropagation. Sodium hypochlorite at 1.2% for 12 minutes can be used for disinfestation of geranium explants, as well as mercury chloride at 0.09 and 0.08% for 12 and 14 minutes, respectively. The leaf explant is more effective for geranium micropropagation using MS medium with 39.8% of its salts. BAP and IAA proportionate the best results for in vitro multiplication and regeneration and number of shoots per explant. For acclimatization the substrates vermiculite and weekly addition of solution with salts of MS medium, coconut dust + Biosafra® (3-12-6) (12 g.L-1) + lime stone (1 g.L-1) and coconut dust + Biosafra® (3-12-6) (12 g.L-1) + lime stone (1 g.L-1) + vermiculite (1:1) can be used. There is a difference in geranium essential oil content and chemical composition between plants obtained from rooted cuttings and micropropagation. / Muitas espécies vegetais sintetizam e acumulam substâncias orgânicas, tais como os óleos essenciais, que podem se tornar alternativa de renda para muitos agricultores. O Pelargonium graveolens L. é uma espécie aromática nativa da África do Sul, popularmente conhecida como gerânio. Seu óleo essencial é amplamente empregado em indústrias de perfumes, cosméticos e fragrâncias, além da aromaterapia. Os métodos convencionais de propagação vegetativa do gerânio não apresentam sucesso considerável, o que tem dificultado a produção de mudas em larga escala. Dessa forma, é importante buscar técnicas alternativas de propagação, como a micropropagação, dessa espécie comercialmente importante. Os objetivos do presente trabalho foram estabelecer um protocolo de micropropagação e aclimatização da espécie, além de verificar o teor e a composição química do seu óleo essencial. Foi utilizado o delineamento inteiramente casualizado. Nos ensaios de micropropagação foram testadas diferentes concentrações e tempos de imersão de hipoclorito de sódio e cloreto de mercúrio, diferentes tipos de explantes e concentrações de sais MS, além de testar os reguladores de crescimento BAP, ANA, cinetina e AIA em diferentes concentrações na ausência e presença de luz. Para o ensaio de aclimatização foram testados diferentes substratos. Por fim, para a análise do óleo essencial foram utilizadas plantas propagadas através da estaquia e micropropagadas. O hipoclorito de sódio a 1,2% por 12 minutos pode ser usado na desinfestação dos explantes de gerânio, assim como o cloreto de mercúrio nas concentrações de 0,09 e 0,08% por 12 e 14 minutos, respectivamente. O explante foliar é o mais eficiente para a micropropagação em meio MS com 39,8% dos sais. Para a multiplicação in vitro o uso de BAP e AIA proporciona os melhores resultados para a regeneração e para o número de brotos por explante e na aclimatização pode-se usar os substratos vermiculita com adição semanal de sais MS, pó de coco + Biosafra® (3-12-6) (12 g.L-1) + calcário (1 g.L-1) e pó de coco + Biosafra® (3-12-6) (12 g.L-1) + calcário (1 g.L-1) + vermiculita (1:1). Há diferença no teor e composição química do óleo de gerânio proveniente de plantas propagadas por estaquia e micropropagadas.