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Previous issue date: 2016-09-30 / CAPES / This study aimed determinate the chemical composition of raw secretions expelled by two species of Aparasphenodon genus from compounds isolation, purification and structural identification. For this, individuals of A. brunoi (n=5) e A. arapapa (n=3) were collected in Ilha da Marambaia, municipality of Mangaratiba (RJ), and in municipality of Ilh?us (BA), respectively. The secretions were obtained by smooth electric stimulus and were translucent and little viscous, with approximated yield of 4 mg of sample/individual. For each species, was realized chromatography of thin layer, the revelator reagent ninhydrin indicated presence of amine groups after violet color appearing. Samples than were submitted with the same retention time (0.88 and 1.44 min) in both analysis, indicating presence of similar compounds in both species. In same way, samples purification realized by semi preparative liquid chromatography of high efficiency presented the same fractions number for both species. The structural determination was made by LC-MS/MS only for A. brunoi and reveled high collagen incidence and other uncharacterized proteins. Biological assays were realized in concentrations of 1?g.mL-1 and 10?g.mL-1 for both species. The assay of cellular viability did not present survival reduction of yeast, such as dosing of lipid peroxidation that did not show reaction with malonaldehyde in all tested cases. In other hand, the assay of mitochondrial damage reveled toxicity in concentration of 10 ?g.mL-1, reducing for discreet form the cellular viability in both samples. We conclude that both species have proteins as the main compound and, in a cellular level, are not harmful in 1 ?g.mL-1, but can promote damage in 10 ?g.mL-1. Besides that, as reveled profiles to both extracts, both chemical and biological assays were very close, the existence of a pattern in chemical composition of cutaneous secretions in amphibians of the same genus can be reinforced, even that species be in distinct environmental conditions. / O presente estudo buscou determinar a composi??o qu?mica das secre??es brutas expelidas por duas esp?cies de Aparasphenodon, a partir do isolamento, purifica??o e identifica??o estrutural destes compostos. Para tal, indiv?duos de A.brunoi (n=5) e A.arapapa (n=3) foram coletados na Ilha da Marambaia, Mangaratiba (RJ) e no munic?pio de Ilh?us (BA), respectivamente. As secre??es foram obtidas por est?mulo el?trico suave e mostraram-se translucidas e pouco viscosas, com rendimento aproximado de 4 mg de amostra/indiv?duo. Para cada esp?cie, foi realizada cromatografia em camada delgada; o reagente revelador ninidrina indicou presen?a de grupos am?nicos ap?s aparecimento da cor violeta. As amostras foram ent?o submetidas ? cromatografia liquida de alta efici?ncia anal?tica, sendo detectados dois picos majorit?rios com mesmo tempo de reten??o (0,88 e 1,44 min) em ambas as an?lises, indicando assim a presen?a de compostos semelhantes nas duas esp?cies. Da mesma forma, a purifica??o das amostras realizada por cromatografia liquida de alta efici?ncia semi-preparativa apresentou o mesmo n?mero de fra??es para ambas as esp?cies. A determina??o estrutural foi determinada por LC-MS/MS apenas para A. brunoi e revelou alta incid?ncia de col?geno e outras prote?nas n?o caracterizadas. Os ensaios biol?gicos foram realizados nas concentra??es de 1 ?g.mL-1 e 10 ?g.mL-1 para ambas as esp?cies. O ensaio de viabilidade celular n?o apresentou redu??o da sobreviv?ncia das leveduras, assim como a dosagem de peroxida??o lip?dica n?o mostrou rea??o com malonalde?do, em todos os casos testados. Por outro lado, o ensaio de dano mitocondrial revelou toxicidade na concentra??o de 10 ?g.mL-1, reduzindo de forma discreta a viabilidade celular, em ambas as amostras. Conclu?mos ent?o que as duas esp?cies possuem prote?nas como seu principal componente e, a n?vel celular, n?o s?o nocivas em 1 ?g.mL-1, mas podem promover danos quando em 10 ?g.mL-1. Al?m disso, como os perfis revelados para ambos os extratos, tanto nos ensaios qu?micos como nos biol?gicos, foram muito pr?ximos, a exist?ncia de um padr?o na composi??o qu?mica das secre??es cut?neas em anf?bios de um mesmo g?nero pode ser refor?ada, ainda que as esp?cies se encontrem em condi??es ambientais distintas.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:jspui/2014 |
| Date | 30 September 2016 |
| Creators | MONKEN, Francine Fran?a |
| Contributors | Chaves, Douglas Siqueira de Almeida, Riger, Cristiano Jorge, Chaves, Douglas Siqueira de Almeida, Salles, Cristiane Martins Cardoso de, Oliveira, Daniela Barros de |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Qu?mica, UFRRJ, Brasil, Instituto de Ci?ncias Exatas |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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