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Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xv, 136 leaves / The chemical and biological properties of the essential oil from Cymbopogon validus were investigated. Hydro-distillation was used to extract the oil from C. validus, the flower-heads, leaves, culms and rhizomes. The percentage oil yields obtained from the plant organs varied from 0.05 to 1.23%, with the greatest concentration found in the flower-heads and rhizomes, 1.23 and 1.12% respectively. A sensory evaluation of the oil revealed that the essential oil was slightly murky, pale yellow in colour, had a strong turpentine-like smell and remained liquid at room temperature. The oxidative stability of C. validus oil was evaluated by determining its Rancimat induction period (negative), peroxide value (60.56 meq/kg), iodine value (84.55), percentage free fatty acids (0.19%) and percentage cholesterol (3.03%). These results indicated that the oil was highly susceptible to oxidation. Chromatographic profiles of the oils from C. validus, as well as the plant organs were generated using gas chromatography-mass spectroscopy (GC-MS). Predominant compounds present in the oil included alpha-Cubebene, Camphene, Geraniol, Limonene, Myrcene, Palmitic acid and Sabinene. C. validus essential oil was also investigated for its antimicrobial (disk diffusion), antioxidant (1, 1-Diphenyl-1-Picrylhydrazyl (DPPH) assay), anti-inflammatory (5-lipoxygenase assay), anti-mosquito properties (insecticidal, larvicidal and repellency assays) and toxicity profile (Brine shrimp and Ames assays). The oil showed poor antimicrobial activity and inhibited the growth of only Gram positive bacteria with a minimum inhibitory concentration (MIC) of 0.0625 (vol/vol) for Bacillus, Micrococcus and Staphylococcus species. The oil also exhibited excellent antioxidant activity, scavenging more than 80% of DPPH free radicals and possesses anti-inflammatory activity (IC50=190 ppm). C. validus oil showed good adulticidal activity (53.7% mortality) and excellent larvicidal (100% mortality) and repellent activity (100% repellency) against Anopheles arabiensis mosquitoes. At high concentrations, the oil was toxic to brine shrimp larvae. However, when diluted it was safe and the minimum inhibitory concentration was 0.0001(vol/vol). The absence of revertant colonies at all essential oil concentrations in the Ames test suggest that the oil is not mutagenic. These results lead the way for exploiting C. validus oil as a multi-functional agent that has antibacterial, anti-oxidant, anti-inflammatory, and anti-mosquito properties.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2007 xv, 136 leaves / The chemical and biological properties of the essential oil from Cymbopogon validus were investigated. Hydro-distillation was used to extract the oil from C. validus, the flower-heads, leaves, culms and rhizomes. The percentage oil yields obtained from the plant organs varied from 0.05 to 1.23%, with the greatest concentration found in the flower-heads and rhizomes, 1.23 and 1.12% respectively. A sensory evaluation of the oil revealed that the essential oil was slightly murky, pale yellow in colour, had a strong turpentine-like smell and remained liquid at room temperature. The oxidative stability of C. validus oil was evaluated by determining its Rancimat induction period (negative), peroxide value (60.56 meq/kg), iodine value (84.55), percentage free fatty acids (0.19%) and percentage cholesterol (3.03%). These results indicated that the oil was highly susceptible to oxidation. Chromatographic profiles of the oils from C. validus, as well as the plant organs were generated using gas chromatography-mass spectroscopy (GC-MS). Predominant compounds present in the oil included alpha-Cubebene, Camphene, Geraniol, Limonene, Myrcene, Palmitic acid and Sabinene. C. validus essential oil was also investigated for its antimicrobial (disk diffusion), antioxidant (1, 1-Diphenyl-1-Picrylhydrazyl (DPPH) assay), anti-inflammatory (5-lipoxygenase assay), anti-mosquito properties (insecticidal, larvicidal and repellency assays) and toxicity profile (Brine shrimp and Ames assays). The oil showed poor antimicrobial activity and inhibited the growth of only Gram positive bacteria with a minimum inhibitory concentration (MIC) of 0.0625 (vol/vol) for Bacillus, Micrococcus and Staphylococcus species. The oil also exhibited excellent antioxidant activity, scavenging more than 80% of DPPH free radicals and possesses anti-inflammatory activity (IC50=190 ppm). C. validus oil showed good adulticidal activity (53.7% mortality) and excellent larvicidal (100% mortality) and repellent activity (100% repellency) against Anopheles arabiensis mosquitoes. At high concentrations, the oil was toxic to brine shrimp larvae. However, when diluted it was safe and the minimum inhibitory concentration was 0.0001(vol/vol). The absence of revertant colonies at all essential oil concentrations in the Ames test suggest that the oil is not mutagenic. These results lead the way for exploiting C. validus oil as a multi-functional agent that has antibacterial, anti-oxidant, anti-inflammatory, and anti-mosquito properties.

Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2016-09-27T22:26:12Z No. of bitstreams: 1 DISSERTA??O Val?ria Silva G?es.pdf: 1629398 bytes, checksum: 6e130737f6d57007b90ea3ae8de5042b (MD5) / Made available in DSpace on 2016-09-27T22:26:12Z (GMT). No. of bitstreams: 1 DISSERTA??O Val?ria Silva G?es.pdf: 1629398 bytes, checksum: 6e130737f6d57007b90ea3ae8de5042b (MD5) Previous issue date: 2015-03-31 / The Lamiaceae family has a great importance in the Brazilian semiarid region due to their species of pharmacological properties. Martianthus leucocephalus (Mart. Ex Benth.) JFB Pastore belongs to the Lamiaceae family and has had the antimicrobial activity of essential oil proven. However, this species lacks pharmacological studies that evaluate its biological potential. Thus, the aim of this study was to evaluate the effect on the central nervous system, the acute toxicity, as well as investigate the antinociceptive and anti-inflammatory of Martianthus leucocephalus extract. The possible mechanism of action of species was also studied. The experiments were performed in the Chemistry Laboratory of Bioactive Natural Products and in the Laboratory of Pharmacology at the State University of Feira de Santana. The effect on the central nervous system was evaluated through Behavioral Pharmacological Screening by Rota Rod test. The acute toxicity was determined using the protocol Organization for Economic Cooperation and Development (OECD - 420/2001). The antinociceptive activity was investigated in the tests of acetic acid, formalin and hot plate. The mechanism of action was determined using the opioid antagonist naloxone in the hot plate model. The study of anti-inflammatory activity was performed using the paw edema induced by carrageenan tests and capillary permeability induced by acetic acid. The results were analyzed by analysis of variance (ANOVA). Martianthus leucocephalus showed no acute toxicity or muscle relaxant effects action on the motor system. The antinociceptive and anti-inflammatory properties were found significantly compared to the control group animals. / A fam?lia Lamiaceae tem uma grande import?ncia na regi?o do semi?rido brasileiro devido ?s suas esp?cies detentoras de propriedades farmacol?gicas. Martianthus leucocephalus (Mart. ex Benth.) J.F.B. Pastore pertence ? fam?lia Lamiaceae e j? teve a atividade antimicrobiana do seu ?leo essencial comprovada. No entanto, a referida esp?cie carece outros de estudos farmacol?gicos que avaliem seu potencial biol?gico. Diante disso, o objetivo desse trabalho foi avaliar o efeito sobre o sistema nervoso central, a toxidade aguda, bem como investigar as atividades antinociceptiva e anti-inflamat?ria de extratos de Martianthus leucocephalus proveniente de cultivo. O poss?vel mecanismo de a??o da esp?cie tamb?m foi estudado. Os experimentos foram realizados no Laborat?rio de Qu?mica de Produtos Naturais e Bioativos e no Laborat?rio de Farmacologia da Universidade Estadual de Feira de Santana. O efeito sobre o sistema nervoso central foi avaliado por meio da Triagem Farmacol?gica Comportamental e pelo teste do Rota Rod. A toxidade aguda foi averiguada utilizando o protocolo Organization for Economic Cooperation and Development (OECD ? 420/2001). A atividade antinociceptiva foi investigada nos testes do ?cido ac?tico, formalina e placa quente. O mecanismo de a??o foi apurado utilizando o antagonista opioide naloxona no modelo da placa quente. O estudo da atividade anti-inflamat?ria foi realizado utilizando os testes do edema de pata induzido por carragenina e da permeabilidade capilar induzida por ?cido ac?tico. Os resultados obtidos foram analisados atrav?s da an?lise de vari?ncia (ANOVA). Martianthus leucocephalus n?o mostrou toxidade aguda nem a??o miorrelaxante sobre o sistema motor. As propriedades antinociceptiva e anti-inflamat?ria foram evidenciadas de forma significativa em rela??o aos animais do grupo controle.

Made available in DSpace on 2016-04-28T20:15:36Z (GMT). No. of bitstreams: 1 Tatiana Xavier de Almeida.pdf: 937180 bytes, checksum: e0cda53e5c0c9829836d98da482b3737 (MD5) Previous issue date: 2010-02-09 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Fungal and mycotoxin contamination in food is one of the greatest problems around the world. Some natural products, such as essential oils, have been shown to be effective inhibitors of fungal growth and production of their toxins. In this study, a total of 40 vegetal extracts were tested for inhibition on Aspergillus section Flavi growth. These extracts were obtained from ten herbs cultivated in Brazil: basil (Ocimum basilicum), cinnamon (Cinnamomum zeylanicum), clove (Eugenia caryophyllata), cumin (Cuminum cyminum), marjoram (Origanum majorana), nutmeg (Myristica fragrans), oregano (Origanum vulgare), rosemary (Rosmarinus officinalis), spearmint (Mentha piperita) and sweet fennel (Pimpinella anisum). Preliminary in vitro tests using the agar diffusion method showed that essential oils were more effective in inhibiting the fungal growth when compared with ethanol and aqueous extracts. Essential oils obtained from oregano and rosemary, clove and cumin ethanolic extracts and the essential hexanic oil of oregano and cinnamon were selected for advanced tests. Differents concentrations of plant material (0, 50, 100, 150, 300 and 600 μg/g) were added to culture medium. A suspension of 103 conidia per inoculation were used. Plates were incubated for ten days while daily measurements of colony diameters were made to obtain growth rate (mm/day) and lag phase (h) of toxigenic strains. The comparison of selected essential oils effect on A. flavus and A. parasiticus showed better inhibition by incorporating oregano essential oil in the culture medium at 600 μg/g concentration. Nevertheless, the hexanic essential oil of oregano showed no statistical difference with the control group, a change occurred in the plant product due to the extraction method. Among the evaluated ethanol extracts, clove had the best result. Aspergillus flavus showed a slight reduction of growth (4.5 to 3.5 mm/day) when compared to control group, but a large increase in the latency period (37 h at a 600 μg/g concentration). Aspergillus parasiticus showed gradual decreases as increased concentrations were assayed, except for the 150 μg/g concentration, and a significant increase on lag phase at the highest concentrations tested (45 and 48 h, respectively). The data presented here are very important for the continued efforts of microbiological control in food, especially when it comes to control fungi. / A contamina??o de alimentos por fungos e micotoxinas constitui um grande problema mundial. Alguns produtos naturais, como por exemplo, ?leos essenciais, s?o demonstrados como potenciais inibidores do crescimento f?ngico, bem como da produ??o de suas toxinas. Neste estudo, um total de 40 extratos de plantas foi avaliado como inibidores do desenvolvimento f?ngico de cepas pertencentes ao g?nero Aspergillus se??o Flavi. Tais extratos foram obtidos a partir de dez ervas, utilizadas especialmente como condimentos e cultivadas no Brasil: alecrim (Rosmarinus officinalis), canela (Cinnamomum zeylanicum), cominho (Cuminum cyminum), cravo-da-?ndia (Eugenia caryophyllata), erva-doce (Pimpinella anisum), hortel? (Mentha piperita), manjeric?o (Ocimum basilicum), manjerona (Origanum majorana), noz moscada (Myristica fragrans) e or?gano (Origanum vulgare). Ensaios preliminares in vitro utilizando o m?todo de difus?o em agar demonstraram que os ?leos essenciais foram os mais efetivos na inibi??o do crescimento f?ngico quando comparados com extratos etan?licos e aquosos. Foram selecionados os ?leos essenciais de or?gano e alecrim, os extratos etan?licos de cravo-da-?ndia e cominho e, os ?leos essenciais hex?nicos de or?gano e canela para ensaios mais aprofundados. Diferentes concentra??es de extratos de plantas arom?ticas (0, 50, 100, 150, 300 e 600 μg/g) foram adicionadas ao meio de cultivo. Utilizou-se uma suspens?o de 103 con?dios por inocula??o. As placas foram incubadas por dez dias enquanto medi??es di?rias do di?metro da col?nia eram realizadas a fim de obter velocidade de crescimento (mm/dia) e fase de lat?ncia (h) das cepas tox?genas. Comparando os ?leos essenciais selecionados, tanto A. flavus quanto A. parasiticus apresentaram uma melhor inibi??o atrav?s da incorpora??o do ?leo essencial de or?gano no meio de cultivo a uma concentra??o de 600 μg/g. Apesar disso, o ?leo essencial hex?nico de or?gano n?o demonstrou qualquer diferen?a estat?stica em rela??o ao grupo controle, havendo ent?o altera??o em algum componente do extrato devido a forma de extra??o. Entre os extratos etan?licos avaliados, o cravo-da-?ndia obteve o melhor resultado. Aspergillus flavus demonstrou uma discreta diminui??o de crescimento (4,5 a 3,5 mm/dia), quando comparado ao grupo controle, mas um grande aumento em sua fase de lat?ncia (37 h a 600 μg/g). Aspergillus parasiticus apresentou uma diminui??o gradativa conforme ocorria o aumento das concentra??es, exceto pela concentra??o de 150 μg/g, e um aumento significativo da fase lag nas maiores concentra??es testadas (45 e 48 h respectivamente). Os dados apresentados aqui s?o de extrema relev?ncia para os constantes esfor?os de controle microbiol?gico em alimentos, principalmente em se tratando de controle f?ngico.

Thesis (MSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Bacterial and fungal pathogens have developed numerous defence mechanisms against antimicrobial chemical agents, and resistance to old and new produced drugs are on the rise. Discovery of natural products derived from plants with diverse chemical structures and novel mechanisms of action to treat these notorious pathogens is a priority. Biotechnology (discussed in Chapter 1) has much to offer as a pharmacological tool and in the general study of medicinal plants. The Genus Salvia (Lamiaceae) has gathered much interest as these plants manufacture a diverse range of secondary metabolites including flavonoids, tannins and terpenoids. Of particular interest are the terpenoids which are largely implicated in the efficacy of Salvia plants as traditional medicines contributing to their pharmacological actions (discussed in Chapter 2). Due to the importance of these plants as herbal remedies, in this study, biotechnological techniques such as tissue culture and Agrobacterium-mediated transformation were applied on Salvia runcinata L.f., a South African medicinal plant, in an attempt to enhance the metabolomic profile and its bioactivity. Like so many other sages, S. runcinata has been used in folk medicine to treat a variety of ailments. Application of biotechnology was viewed as an important value adding platform for this species, assisting with its commercialisation for the cosmeceutical and pharmaceutical industries. Therefore the study had three foci: (1) to determine the seed germination behaviour and optimal conditions for micropropagation; (2) to develop a protocol that would be efficient whilst being simple for genetic transformation; and lastly, (3) to conduct phytochemical studies on in vitro generated S. runcinata transgenic hairy root and in vitro organ cultures by comparing these to glasshouse plants as potential therapeutic sources of natural compounds used in the treatment of infections in plants and humans. Data generated is thus summarised in three research chapters and Chapter 3 describes the formulated procedures assisting with in vitro seed germination and micropropagation of S. runcinata. The efficacy of smoke and scarification treatments for germination improvement was initially tested coupled to the evaluation of different hormonal combinations and different explant types which would aid with inducing adventitious shoot formation in vitro. The most effective germination treatment proved to be a 3 min exposure of seeds to 25% (w/v) H2SO4 combined with a concentration of 10-5 M smoke solution, resulting to more than 80% germination. Shoot proliferation was significantly higher using nodal explants with the addition of 4.43 μM BA. The protocol established in this part of the study is viable for large scale commercial production of S. runcinata as it would yield 1296 to 46656 viable plants in 4 to 6 months from one nodal explant. Micropropagation was applied also as a pre-emptive measure to ease pressure on the wild plants as the demand for S. runcinata is anticipated to increase due to its growing economic value as it is one of two South African sages with epi-α-bisabolol that is sought after by the pharmaceutical and cosmeceutical industries. This makes the protocol developed in this part of the study suitable for ex situ conservation of S. runcinata plantlets. Evaluations on the transgene transfer capacities of two different agropine strains (A4T and LBA 9402) of Agrobacterium rhizogenes to induce hairy root cultures of S. runcinata explants on nodal and leaf explants were conducted (reported in Chapter 4). Hairy roots formed 3 to 4 weeks after inoculation of the explants and these agropine strains showed different abilities for genetic transformation with the LBA 9402 strain producing significantly more roots on each explant compared to the A4T strain (P=0.0075). However, none of the LBA 9402 derived clones and only 2 clones generated through A4T transformation survived subculturing. The polymerase chain reaction (PCR) and reverse transcriptase-polymerase chain reaction (RT-PCR) confirmed the presence and transcription (respectively) of rol A, rol B, rol C and ags genes which are mobilised from the transfer-DNA (T-DNA) fragment of the root-inducing (Ri) plasmid of A. rhizogenes to the plant genome during transformation. The two A4T clones, termed here A4T3 and A4T5, were stably transformed, Southern blot analysis using rol A as a probe further validated the integration of one copy of the rol A gene. Transformed hairy roots, untransformed roots from tissue cultured plants, tissue culture-derived plants and glasshouse-grown plants were profiled for secondary metabolites by thin layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS) in Chapter 5. In this part of the study, it is clear that the use of tissue culture as a propagation system did not negatively affect the volatile compound profile of S. runcinata and plants had a similar essential oil content to that reported by Kamatou et al. (2008), leading to a conclusion that in vitro plants maintained their biochemical integrity even under an alternative micro-controlled environment. Similarly to others, Ri-transformation was explored as an avenue to alter secondary metabolism creating inter-clonal variation. Transformed clones were distinguishable, displaying more of some primary metabolites including sucrose, galactose, sorbose and fructose than the leaf extracts. With the current GC-MS methods used, this clear distinction was not obvious at the secondary metabolite level. In general, solvent extracts (acetone and methanol:dichloromethane (MetOH: DCM) (1:1 v/v) exhibited good to moderate antibacterial activity with the minimum inhibitory concentration (MIC) values ranging from 0.39 to 0.78 mg ml-1. However, in vitro plant cultures were the most potent against two Gram-negative bacterial strains: Escherichia coli (ATCC 11775) and Klebsiella pneumoniae (ATCC 13883), and two Gram-positive bacterial strains: Bacillus subtilis (ATCC 6051) and Staphylococcus aureus (ATCC 12600). The hairy root extracts did not show any activity against fungi, Fusarium subglutinans (MRC 0115) and Fusarium proliferatum (MRC 6908). Micropropagation therefore proves to be an interesting avenue for commercial production of S. runcinata, supplying plants with an improved pharmacological activity. Hence the biotechnological approach applied here is a viable strategy for the production of medicinal bioactives from S. runcinata. / AFRIKAANSE OPSOMMING: Bakterieë en fungi patogene het baie verskeie meganismes ontwikkel teen antimikrobiese chemiese agente, en weerstand teen ou en nuwe chemise stowwe is besig om te vergroot. Daarom is dit belangrik om natuurlike plantaardige produkte met diverse chemiese strukture en unieke werkings meganismes te ontdek waarmee hierdie berugte patogene beveg kan word. Biotegnologie (wat in Hoofstuk 1 bespreek word) kan gebruik word as 'n farmakologiese hulpmiddel in die algemene studie van plante. Die Klas (Genus) Salvia (Lamiaceae) het al baie aandag getrek aangesien hierdie plante 'n wye reeks sekondêre metaboliete vervaardig wat flavonoïede, tanniene en terpenoïede insluit. Veral van belang is die terpenoïde wat betrokke is by die doeltreffendheid van die Salvia plante as tradisionele medisyne, aangesien dit bydra tot hulle farmalogiese aksie (wat in Hoofstuk 2 bespreek word). Aangesien hierdie plante sulke belangrike kruie is, word daar in hierdie studie, biotegnologiese tegnieke soos die kweek van weefsel en Agrobacterium-bemiddelde transformasie op Salvia runcinata L.f. toegepas om die metabologiese profiel en die bioaktiwiteit daarvan te verbeter. Soos baie van die salies is S. runcinata tradisioneel dikwels gebruik om allerhande siektetoestande te behandel. Die toepassing van biotegnologie word beskou as 'n belangrike manier om waarde by te voeg sodat hierdie plant kommersieei deur die kosmetiese en farmakeutiese bedrywe gebruik kan word. Daarom is daar op drie dinge gefokus: (1) die ontkiemings gedrag van saad en die optimale toestande vir mikrovoortplanting (2) die ontwikkeling van protokol wat eenvoudig maar doeltreffend is vir genetiese transformasie, en die (3) fito-chemise studies op in vitro genereerde S. runcinata transgeniese harige wortels en in vitro orgaan kwekings deur om hulle te vergelyk met kweekhuis plante as potentiële terapeutiese bronne van natuurlike samestellings vir die behandeling van infeksies in beide plante en mense. Die data wat gegenereer is, is opgesom in drie hoofstukke, en in Hoofstuk 3 word die prosedures wat gebruik word in die in vitro saad ontkieming en die mikro voortplanting van S. runcinata, bespreek. Die doeltreffendheid van rook en skarifikasie behandeling vir die verbetering van ontkieming is eers getoets en gekoppel aan die evaluering van verskillende hormoonkombinasies en verskillende eksplant tipes wat lei tot die formasie van uitloopsels in vitro. Daar is gevind dat die effektiefste behandeling vir ontkieming, 'n 3-minuut blootstelling van saad aan 25% (w/v) H2SO4 gekombineer met 'n konsentrasie 10-5 M rook oplossing is. Dit het gelei tot meer as 80% ontkieming. Daar was baie meer uitloopsels toe nodale eksplante gebruik is met die byvoeging van 4.43 μM BA. Die proktokol wat hier gevestig is, kan op groot skaal gebruik word vir die kommersiële produksie van S. runcinata, want 1296 tot 46656 lewensvatbare plante kan binne 4 ot 6 maande van een nodale eksplant gemaak word. Mikro voortplanting is toegepas as 'n voorkomende maatreel om die druk op die natuur te verminder omdat daar verwag word dat die vraag na S. runcinata sal toeneem na gelang die groeiende ekonomiese waarde daarvan toeneem. Dit is een van twee Suid-Afrikaanse salies met epi-α-bisabolol wat deur die farmakeutiese en die kosmetiese bedrywe gebruik word. Dit beteken dat die protokol wat hier ontwikkel is, geskik is vir die ex situ bewaring van S. runcinata plante. Die transgeen oordrag van twee verskillende agropien tipes (A4T and LBA 9402) van Agrobacterium rhizogenes is geevalueer (en in Hoofstuk 4 beskryf). Harige wortels het 3 tot 4 weke na die inenting van die eksplante gevorm en hierdie agropien tipes het verskillende vermoëns vir genetiese transformasie getoon, met die LBA 9402 tipe wat baie meer wortels op elke eksplant voorgebring het in vergelyking met die A4T tipe (P=0.03116). Geen van die LBA 9402-afgeleide klone en slegs 2 klone wat deur A4T transformasie genereer is, het oorleef. The polimerase ketting reaksie (PCR) en die teenoorgestelde trenskriptasie-polimerase (RT-PCR) ketting reaksie het die teenwoordigheid en transkipsie (onderskeidelik) van rol A, rol B en rol C en ags gene, wat oorgedra word deur die oordrag DNA (T-DNA) fragment van die wortel induserende (Ri) plasmied van A. rhizogenes na die plant genoom tydens transformasie, bevorder. A4T klone, hier A4T3 and A4T5 genoem, is stabiel transformeer. Southern blot ontleding het met die gebruik van rol A, die integrasie van een kopie van die rol A geen, bevestig. In Hoofstuk 5 is transformeerde harige wortels, ongetransformeerde wortels van weefsel gekweekte plante, weefsel gekweekte plante, en kweekhuis plante deur dun-laag chromatografie (TLC) en gas-chromatografie-massa spektrometrie (GC-MS) geprofiel vir sekondêre metaboliete. In hierdie deel van die studie is dit duidelik dat die gebruik van weefsel kwekery as 'n voortplantsisteem nie 'n negatiewe effek gehad het op die vlugtige samestelling profiel van S. runcinata nie en dat plante 'n sootgelyke essentiële olie inhoud het as wat deur Kamatou et al. (2008) bevind is. Dit lei tot die gevolgtrekking dat in vitro plante hulle biochemiese integriteit behou selfs onder alternatiewe mikro-beheerde omgewings. Ri-transformasie is ondersoek as 'n manier om sekondêre metabolisme te verander om interkloon variasie te skep. Getransformeerde klone kon uitgeken word, aangesien dit meer primêre metaboliete soos sukrose, galaktose en fruktose insluit as die blaar ekstrakte. Hierdie verskil was nie met die huidige GC-MS metodes so duidelik sigbaar op die sekondêre metabolitiese vlak nie. Oor die algemeen toon ekstraksie met asetoon en methanol dichlorometaan (MetOH: DCM) (1:1 v/v) goeie tot gemiddelde antibakteriese aktiwiteit met die minimum remmende konsentrasie (MIC) waardes van 0.39 tot 0.78 mg ml-1. Die in vitro plant kulture het egter sterker weerstand gebied teen twee Gram-negatiewe bakteriese tipes: Escherichia coli (ATCC 11775) en Klebsiella pneumoniae (ATCC 13883), en teen twee Gram-positiewe bakteriese tipes: Bacillus subtilis (ATCC 6051) en Staphylococcus aureus (ATCC 12600). Die harige wortel ekstrakte het geen aktiwiteit teen die swamme, Fusarium subglutinans (MRC 0115) en Fusarium proliferatum (MRC 6908) getoon nie. Mikro-voortplanting is dus 'n interessante manier om S. runcinata kommersieel te produseer aangeien die plante verbeterde farmalogiese aktiwiteit toon. Die biotegnologiese benadering wat hier toegepas word, is 'n praktiese strategie vir die produksie van geneesmiddels van S. runcinata.

O óleo essencial de Citrus latifolia Tanaka (limão Tahiti) possui grande valor comercial e aplicações em diversos tipos de indústrias. A extração enzimática de óleos vegetais fixos aumenta o rendimento, consome pouca energia e causa um mínimo impacto ambiental, porém poucos estudos avaliaram o seu emprego para a extração de óleos essenciais de plantas aromáticas. Em vista disso, este trabalho teve como objetivo contribuir para o desenvolvimento de uma tecnologia enzimática para a extração do óleo essencial de cascas de C. latifolia Tanaka, além de avaliar a percentagem de compostos bioativos, como cumarinas, nos resíduos dos processos de extração. Para tanto, foram realizados pré-tratamentos enzimáticos nas cascas, através de um complexo constituído principalmente por celulases, caracterizado em papel filtro (FPA), e posterior extração do óleo por hidrodestilação. Nos três primeiros experimentos foram avaliados o efeito da carga enzimática (5, 7 e 12 FPA/g de substrato), o efeito do tempo de pré-tratamento enzimático (1, 2 e 3 horas) e o efeito do pré-tratamento em cascas trituradas sobre o rendimento e a composição do óleo essencial obtido por 1 hora de hidrodestilação. Em outro experimento foi avaliada a possibilidade de redução no tempo de hidrodestilação através do pré-tratamento enzimático, utilizando-se cargas de 5, 7 e 12 FPA/g em cascas trituradas, calculando-se o rendimento em óleo durante a hidrodestilação em diferentes intervalos de tempo (10, 15, 20, 30, 40, 50 e 60 minutos). As cumarinas foram extraídas com hexano dos resíduos líquidos e sólidos das hidrodestilações através de extração líquido-líquido e soxhlet, respectivamente. As análises dos compostos dos óleos essenciais e das cumarinas foram feitas por cromatografia gasosa (GC) e cromatografia gasosa acoplada à espectrometria de massas (GC/MS). Nos três primeiros experimentos descritos não foram verificadas diferenças estatísticas entre os rendimentos em óleo das amostras pré-tratadas enzimaticamente e os controles, demonstrando que o pré-tratamento enzimático não alterou o rendimento em óleo obtido por 1 hora de hidrodestilação. Porém, a análise dos componentes dos óleos permitiu observar um aumento nas proporções de alguns compostos oxigenados (p<0,05), responsáveis pela qualidade do aroma. No experimento onde foi avaliada a diminuição do tempo de hidrodestilação, verificou-se que o pré-tratamento com carga de 5 FPA/g atingiu o rendimento máximo em torno de 30 minutos do processo de destilação, enquanto que no controle o rendimento máximo foi em torno de 50 minutos. Utilizando-se a carga de 7 FPA/g, o rendimento máximo foi atingido em torno de 20 minutos, o mesmo ocorrendo com a carga de 12 FPA/g, enquanto que no controle foi em torno de 40 e 50 minutos, respectivamente. Estes resultados indicam que o pré-tratamento enzimático reduz o tempo de hidrodestilação apresentando vantagens do ponto de vista de custo energético e de integridade dos componentes do óleo, visto a redução do tempo de exposição do material vegetal à temperatura de ebulição da água, preconizando as bases para um processo tecnológico que envolve pré-tratamento enzimático associado à hidrodestilação para ser empregado na obtenção de óleo essencial de C. latifolia Tanaka. Em relação às cumarinas, os resultados obtidos indicaram que a percentagem da maioria desses compostos não foi alterada quando as amostras sofreram pré-tratamento enzimático. Entretanto, propõe-se que os resíduos gerados pela hidrodestilação podem ser aproveitados e utilizados como fonte de cumarinas. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2015-09-21T19:26:39Z No. of bitstreams: 1 Dissertacao Juliane Cassini.pdf: 948341 bytes, checksum: 92bfa933404cea773a26604ae93106f9 (MD5) / Made available in DSpace on 2015-09-21T19:26:39Z (GMT). No. of bitstreams: 1 Dissertacao Juliane Cassini.pdf: 948341 bytes, checksum: 92bfa933404cea773a26604ae93106f9 (MD5) / Citrus latifolia Tanaka (Tahiti lime) essential oil has a high commercial value and a large application in different industries. The enzymatic extraction of vegetable oils increases yields extraction, energy efficient and cause minimal environmental impact, but few studies have evaluated its use for the extraction of essential oils from aromatic plants. As a result, this study aimed to contribute to the development of enzymatic technology for extraction of essential oil from peels of C. latifolia Tanaka, and assess the proportion of bioactive compounds such as coumarins, in wastes extraction processes. To do so, it was performed enzymatic pretreatment of the peels, through a complex composed primarily of cellulases, characterized in filter paper (FPA), and subsequent extraction of oil by hydrodistillation. In the first three experiments were evaluated the effect of enzyme loading (5, 7 and 12 FPA/g substrate), the effect of time of enzymatic pretreatment (1, 2 and 3 hours) and the effect of pretreatment on crushed peels, on yield and composition of essential oil obtained by 1 hour hydrodistillation. In another test, it was evaluated the possibility of reducing the time hydrodistillation by enzymatic pretreatment, using loads of 5, 7 and 12 FPA/g of crushed peels, calculating the oil yield during the hydrodistillation at different intervals time (10, 15, 20, 30, 40, 50 and 60 minutes). The coumarins were extracted from the liquid and solid wastes from hydrodistillations with hexane through liquid-liquid extraction and soxhlet, respectively. The analysis of the compounds of essential oils and coumarins were made by gas chromatography (GC) and gas chromatography-mass spectrometry (GC / MS). In the first three experiments described not statistical differences were found between the oil yields of enzymatically pretreated samples and controls, demonstrating that the enzymatic pretreatment did not affect the oil yield obtained by 1 hour hydrodistillation. However, the analysis of the oil components allowed to observe an increase in the proportions of some oxygenated compounds (p < 0,05), responsible for the quality of aroma. In the experiment where was evaluated the short-time hydrodistillation, it was found that pretreatment with a load of 5 FPA/g reached the maximum yield of about 30 minutes from the distillation process, while in the control the maximum yield was about 50 minutes. Using the load of 7 FPA/g, the maximum yield was reached in about 20 minutes, the same happened with the load of 12 FPA/g, while in the control was around 40 and 50 minutes, respectively. These results indicate that enzymatic pretreatment reduces the time hydrodistillation with advantages in terms of cost efficiency and integrity of the oil components, since the reduction in exposure time of the plant material at a temperature of boiling water, pointing out the foundation for a technological process that involves enzyme pretreatment associated with hydrodistillation to be employed in obtaining essential oil of C. latifolia Tanaka. For the coumarins, the results indicated that the percentage of most of these compounds was not altered when the samples were enzymatically pretreated. However, it is proposed that the waste generated by hydrodistillation can be recovered and used as a source of coumarins.

Cunila spicata Benth. é uma planta aromática e medicinal utilizada popularmente no sul do Brasil como béquica, peitoral e sudorífera, e que apresenta comprovada atividade antiviral e anticonvulsiva. C. spicata é uma das 12 espécies sul-americanas do gênero Cunila (Lamiaceae) sendo encontrada em banhados e borda de mata de galeria no sul do Brasil, Argentina e Uruguai. Dentro deste contexto, no presente trabalho foi avaliada a variabilidade genética e química de populações de C. spicata coletadas nas regiões nordeste e sudeste do Rio Grande do Sul, com vistas a formação de bancos de germoplasma, determinação de estratégias de preservação e uso sustentável. Os marcadores ISSR e RAPD mostraram-se eficientes na determinação de variabilidade genética desta espécie, permitindo a identificação de todas as populações e indivíduos avaliados. As populações de C. spicata caracterizam-se como grupos geneticamente estruturados. Entretanto, não foi constatada formação de agrupamentos dentro da espécie, nem relação entre as distâncias genéticas e geográficas, ou entre as populações originarias das duas regiões geográficas amostradas. Por outro lado, a análise da composição dos óleos essenciais das 10 populações permitiu separar as mesmas em dois quimiotipos caracterizados por elevada concentração de α-terpineol/limoneno, e linalol, respectivamente. As populações da região sudeste enquadraram-se dentro do quimiotipo linalol, enquanto as da região nordeste no quimiotipo α-terpineol/limoneno. A relação geográfica e química é indicativa da existência de pressão de seleção quanto ao tipo de óleo essencial da região de ocorrência. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2016-12-01T16:00:30Z No. of bitstreams: 1 Dissertacao Marcos Roberto Albuquerque.pdf: 1631570 bytes, checksum: d32cbb1d10de9a5f37eb7dd664ddc20d (MD5) / Made available in DSpace on 2016-12-01T16:00:30Z (GMT). No. of bitstreams: 1 Dissertacao Marcos Roberto Albuquerque.pdf: 1631570 bytes, checksum: d32cbb1d10de9a5f37eb7dd664ddc20d (MD5) Previous issue date: 2016-12-01 / Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq. / Cunila spicata Benth. is a medicinal and aromatic plant popularly used in South Brazil as bequic, expectorant and sudiriferous, and that has confirmed antiviral and anti-convulsive activities. C. spicata is one of the 12 South-American species of the genus Cunila( Lamiaceae) been currently found in swamps and borders of gallery forests in South Brazil, North Argentina and Uruguay. In this context, the present work aimed to evaluate the genetic and chemical variability of C. spicata populations collected at the northeast and southeast regions of Rio Grande do Sul, to subside the construction of a germplasm bank, to determine conservation strategies, and to develop a sustainable use of this species. ISSR and RAPD markers proved to be efficients for the determination of the genetic variability of this species, allowing the identification of all the populations and individuals. C. spicata populations were characterized as genetically structured groups. However, clusters within the species were not evident, and no relations were detected between geographic and genetic distances, or between populations of the two regions. Conversely, the analysis of essential oil composition allowed the populations in two chemotypes, characterized by high concentration of α-terpineol/limonene, and linalool, respectively. The populations of the southeast region belong to the linalool chemotype, and those from the northeast region to the a-terpineol/limonene chemotype. This geographical and chemical relation is indicative of different selection pressures in the two regions.

Orientador: Lauro Euclides Soares Barata / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-10T11:52:46Z (GMT). No. of bitstreams: 1 Lupe_FernandaAvila_M.pdf: 1167669 bytes, checksum: 16d83b8cd46e624a76028cf31d1de6f3 (MD5) Previous issue date: 2007 / Resumo: O presente trabalho envolveu o estudo da composição química dos óleos essenciais de cinco plantas aromáticas Aeollanthus suaveolens, Aniba canelilla, Aniba rosaeodora, Lippia alba e Ocimum basilicum por Cromatografia Gasosa acoplada a Espectrometria de Massas (CG-EM). Amostras destas plantas - óleo essencial, planta fresca, seca e resíduo da extração do óleo essencial - foram extraídas por Microextração em Fase Sólida em Espaço confinado (HS-SPME) e analisadas por CG-EM. Os óleos foram extraídos por arraste a vapor. Os óleos essenciais ricos em linalol de A. rosaeodora (madeira e folhas), L. alba e O. basilicum foram avaliados por CG-EM com coluna HP-5 e a composição dos enantiômeros de linalol determinada usando coluna quiral. Com o resultado dessa comparação, foi avaliada a substituição do óleo essencial da madeira de A. rosaeodora pelos óleos essenciais estudados. Um estudo sazonal de L. alba para definir as concentrações e quiralidade do linalol encontrado foi feito com seis coletas. As principais moléculas dos óleos essenciais e/ou extratos de A. canelilla (1-nitro-2-feniletano e metileugenol) e A. suaveolens (massoia lactona) foram isoladas utilizando técnicas de fracionamento como: destilação, hidrodestilação, extração ácido-base, cromatografia em coluna e cromatografia em camada delgada preparativa, e identificadas por RMN de H e C. A semi-síntese da massoia lactona foi feita a partir de uma molécula comercial (d-decalactona). Ensaios biológicos - antifúngico e anti bacteriano - foram realizados pela técnica de difusão com discos com amostras de A. canelilla e A. suaveolens / Abstract: This work involved the chemical composition study of the following five aromatics plants Aeollanthus suaveolens, Aniba canelilla, Aniba rosaeodora, Lippia alba and Ocimum basilicum using Gas Chromatography / Mass Spectrometry (GC-MS). Samples of these plants - essential oils, fresh and dried plants as well as extraction residues - were extracted using Headspace Solid Phase Microextraction (HS-SPME) and analyzed with GC-MS. The oils were extracted by steam distillation. The essential oils of A. rosaeodora (wood and leaves), L. alba and O. basilicum rich in linalool were evaluated by GC-MS using HP-5 column, while the linalool enantiomer composition using a chiral column. From the results of this comparison the replacement of A. rosaeodora essential oil from its wood by the plants listed above was evaluated. A seasonal study of L. alba to define linalool concentrations and chirality was made using six different samples. The main molecules of the essential oils and/or extracts of A. canelilla (1-nitro-2-phenyethane and methyl eugenol) and A. suaveolens (massoia lactone) were isolated using separation techniques as distillation, hydrodistillation, acid-base extraction, column chromatography and preparative thin layer chromatography, and identified by NMR H and C. The semi-synthesis of the massoia lactone was obtained from a commercial molecule (d-decalactone). Biological assays - anti-fungal and antibacterial - with samples of A. canelilla and A. suaveolens were conducted using the diffusion technique in discs / Mestrado / Quimica Organica / Mestre em Química

Cunila spicata Benth. é uma planta aromática e medicinal utilizada popularmente no sul do Brasil como béquica, peitoral e sudorífera, e que apresenta comprovada atividade antiviral e anticonvulsiva. C. spicata é uma das 12 espécies sul-americanas do gênero Cunila (Lamiaceae) sendo encontrada em banhados e borda de mata de galeria no sul do Brasil, Argentina e Uruguai. Dentro deste contexto, no presente trabalho foi avaliada a variabilidade genética e química de populações de C. spicata coletadas nas regiões nordeste e sudeste do Rio Grande do Sul, com vistas a formação de bancos de germoplasma, determinação de estratégias de preservação e uso sustentável. Os marcadores ISSR e RAPD mostraram-se eficientes na determinação de variabilidade genética desta espécie, permitindo a identificação de todas as populações e indivíduos avaliados. As populações de C. spicata caracterizam-se como grupos geneticamente estruturados. Entretanto, não foi constatada formação de agrupamentos dentro da espécie, nem relação entre as distâncias genéticas e geográficas, ou entre as populações originarias das duas regiões geográficas amostradas. Por outro lado, a análise da composição dos óleos essenciais das 10 populações permitiu separar as mesmas em dois quimiotipos caracterizados por elevada concentração de α-terpineol/limoneno, e linalol, respectivamente. As populações da região sudeste enquadraram-se dentro do quimiotipo linalol, enquanto as da região nordeste no quimiotipo α-terpineol/limoneno. A relação geográfica e química é indicativa da existência de pressão de seleção quanto ao tipo de óleo essencial da região de ocorrência. / Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPq. / Cunila spicata Benth. is a medicinal and aromatic plant popularly used in South Brazil as bequic, expectorant and sudiriferous, and that has confirmed antiviral and anti-convulsive activities. C. spicata is one of the 12 South-American species of the genus Cunila( Lamiaceae) been currently found in swamps and borders of gallery forests in South Brazil, North Argentina and Uruguay. In this context, the present work aimed to evaluate the genetic and chemical variability of C. spicata populations collected at the northeast and southeast regions of Rio Grande do Sul, to subside the construction of a germplasm bank, to determine conservation strategies, and to develop a sustainable use of this species. ISSR and RAPD markers proved to be efficients for the determination of the genetic variability of this species, allowing the identification of all the populations and individuals. C. spicata populations were characterized as genetically structured groups. However, clusters within the species were not evident, and no relations were detected between geographic and genetic distances, or between populations of the two regions. Conversely, the analysis of essential oil composition allowed the populations in two chemotypes, characterized by high concentration of α-terpineol/limonene, and linalool, respectively. The populations of the southeast region belong to the linalool chemotype, and those from the northeast region to the a-terpineol/limonene chemotype. This geographical and chemical relation is indicative of different selection pressures in the two regions.