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Previous issue date: 2016-02-29 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Melocactus zehntneri (Britton & Rose) ? uma das muitas esp?cies vegetais de valor terap?utico que se encontra amea?ada pelas atividades humanas sobre o seu habitat. Ao considerar as rela??es etnofarmacol?gicas e quimiotaxon?micas, uma vez que M. zehntneri pertence ? fam?lia Cactaceae, os constituintes qu?micos da esp?cie se tornam atrativos especialmente no que tange os alcaloides isoquinol?nicos, classe de extremo interesse farmacol?gico enquanto mol?culas citot?xicas. Acredita-se que agregando valor cient?fico ? esp?cie novas pol?ticas p?blicas de conserva??o podem ser desenvolvidas. Este trabalho tem como objetivos avaliar a extra??o de alcaloides de M. zehntneri por macera??o, ultrassom (EAU), ultrassom com liquido i?nico (EAU-liquido i?nico) e extra??o por fluido supercr?tico (EFS) tendo como metas um processo sustent?vel e a valoriza??o do bioma Caatinga. Al?m disso, foram realizados experimentos de citotoxicidade com diferentes concentra??es de fra??es enriquecidas em alcaloides frente ?s linhagens HeLa e C33a respons?veis por carcinomas cervicais. As condi??es de extra??o de macera??o [tempo 24, 48 e 72 horas], EAU [amplitude (20%, 27%, 33% e 40%); tempo (15, 30, 45 e 60 minutos) e propor??o droga:solvente (1:10, 1:20 e 1:40, p/v)], EAU-liquido i?nico [concentra??o (0 mol/mL; 0,05 mol/mL; 0,1 mol/mL; 0,2 mol/mL e 0,3 mol/mL)] e EFS [press?o (100, 200 e 300 bar) e temperatura (35? C, 45? C e 55? C)] foram otimizadas para garantir o m?ximo de efici?ncia de extra??o. O rendimento global, Cromatografia em Camada Delgada (CCD) e espectrofotometria foram usadas como ferramentas de an?lises para selecionar as melhores condi??es de extra??o em cada t?cnica. Dessa forma, as amostras obtidas a partir de partes a?reas da planta e extra?das por macera??o (72 horas), EFS (300 bar e 55? C) EAU e EAU-liquido i?nico (27% de amplitude, 45 minutos e propor??o de 1:10, p/v) foram selecionadas e um m?todo em CLAE-UV/DAD desenvolvido para compara??o de perfis. Nessa t?cnica, ?rea e intensidade de absor??o simult?nea em 254 nm e 279 nm dos picos no cromatograma foram assumidos como correspondentes a alcaloides. Os resultados dos cromatogramas sugerem que a EFS ofereceu diferen?as qualitativas e uma maior efici?ncia para todos os compostos, quando comparado com as demais t?cnicas de extra??o; em contrapartida, para a macera??o, EAU e EAU-l?quido i?nico n?o foram observadas diferen?as qualitativas. Na atividade biol?gica foi observada a??o citot?xica frente ? linhagem C33a nas fra??es CHCl3 e BuOH obtidas de partes a?reas e ra?zes; ao passo que, essa atividade em linhagem HeLa apenas foi observada em fra??es clorof?rmicas obtidas de partes a?reas e ra?zes. A aplica??o de t?cnicas sustent?veis e os resultados experimentais apresentados no trabalho agregaram valor cient?fico para a esp?cie de Melocactus zehntneri. Assim, esperamos que essa contribui??o auxilie no desenvolvimento de pol?ticas p?blicas para preserva??o dessa esp?cie e valoriza??o do bioma Caatinga. / Melocactus zehntneri (Britton & Rose) is popularly known as ?coroa-de-frade? and it?s one of many plant species with therapeutic value which is threatened by human activities on their habitat. Taking into account ethnopharmacoly and chemotaxonomy the chemical constituents in M. zehntneri belong to the Cactaceae family become attractive especially regarding the isoquinoline alkaloids, which display pharmacological interest as cytotoxic molecules. It was believed that adding scientific value to the species new public policies for conservation could be developed. This work aimed to evaluate the extraction of alkaloids M. zehntneri by maceration, ultrasound (UAE), ultrasound with ionic liquid (UAE-IL) and supercritical fluid extraction (SFE) with the goals of achieve a sustainable process and the appreciation of the Caatinga biome. Furthermore, cytotoxicity experiments were performed with different concentrations of plant fractions against C33a and HeLa, cell lines responsible for cervical carcinomas. The maceration [time (24, 48 and 72 h)], UAE [power (20, 27, 33 and 40%); time (15, 30, 45 and 60 minutes) and liquid-solid ratio (1:10, 1:20, 1:40, w/v)], UAE-IL [concentration (0 mol/mL; 0.05 mol/mL; 0.1 mol/mL, 0.2 mol/mL and 0.3 mol/mL)] and SFE extraction conditions [pressure (100, 200 and 300 bar) and temperature (35, 45 and 55 ?C)] were optimizated to ensure maximum extraction efficiency. The global yield, Thin Layer Chromatography (TLC) and spectrophotometric analysis were used as tools to select the best extraction conditions for each technique. Thus, samples obtained from the aerial parts of plant and extracted by maceration (72h), SFE (300 bar and 55 ?C), UAE (27%, 45 min and 1:10, w/v) were selected and a HPLC-UV/DAD methodology developed to compare the chromatographic profiles of these samples. In this technique, the selection of peak in the chromatogram probably corresponding to alkaloids was performed based on the spectrum area and intensity of simultaneous absorption at 254 nm and 279 nm. In this technique, selecting the peak in the chromatogram corresponding to the probable alkaloid was performed based on the spectrum area and intensity of simultaneous absorption at 254 nm and 279 nm peaks. The chromatogram results propose qualitative differences and a higher efficiency in SFE when compared with other extraction techniques; on the other hand, in maceration, UAE and UAE-IL qualitative differences were not observed. In biological activity was observed cytotoxic action against the cell line C33a in CHCl3 and BuOH fractions obtained from the aerial parts and roots; whereas this activity in cell line HeLa was only observed in CHCl3 fractions obtained from the aerial parts and roots. The application of sustainable techniques and experimental results presented in the scientific work aggregated value for the Melocactus zehntneri.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/21491 |
| Date | 29 February 2016 |
| Creators | Brand?o, Gustavo Henrique Azevedo |
| Contributors | 00104164042, Arag?o, Cicero Fl?vio Soares, 62001728468, Barbosa, Euz?bio Guimar?es, 94373531153, Danielski, Leandro, 93610050900, Pereira, Camila Gambini, Giordani, Raquel Brandt |
| Publisher | PROGRAMA DE P?S-GRADUA??O EM CI?NCIAS FARMACEUTICAS, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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