Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFPB |
Texto Completo: | https://repositorio.ufpb.br/jspui/handle/tede/6840 |
Resumo: | Propolis is an heterogeneous mixture of substances, produced by melliferous bees from plants in close proximity with the bee´s hives. It is usually classified according to its botanical origin and chemical composition that vary according to geographical origin. Studies have shown that red propolis have physicochemical and biological properties that are diverse from other propolis types. There is a scarcety of studies with Brazilian red propolis, and no study with the red propolis from Igarassu (PE, Brazil), and thus its botanical origin, chemical characterization, and pharmacological activities have not been determined. The aim of this study was to perform a bioassay-guided fractionation of two propolis samples, collected from Igarassu, in order to determine the substances responsible for its antimicrobial, antioxidant, antileukemic and leishmanicidal activities, as well as its palynological profile. The samples of propolis were collected in Igarassu-PE, in natura, in May 2012 in two apiaries. They were submitted to palynological analyses to identify the pollen types and their frequency in the samples. Then, samples in natura were powdered, extracted with 96% ethanol and concentrated to obtain the crude ethanol extract. This extract was dissolved in a mixture of methanol:water (1:1, v/v) and partitioned with hexane and ethyl acetate, obtaining the hexane, ethyl acetate and hydroalcoholic fractions. The acetate fraction was submitted to column chromatography with Sephadex LH20 for isolation of the compounds. The propolis in natura was subjected to preliminary phytochemical screening. The ethanol extract was evaluated for identification of compounds by HPLC-MS. The propolis in natura, ethanol extract, hexane fraction, ethyl acetate and hydroalcoholic were submitted to HPLC analysis for identification and quantification of phenolic compounds. A total of 28 different pollen types belonging to 16 families and 30 genera were found in the samples. The most frequent were: Serjania and Dalbergia (sample 1 April 2011), Fabaceae 1 and Dalbergia (sample 2 April 2011), Stigmaphyllon (sample 1 and 2 May 2012). It was found the presence of flavonoids and steroids in propolis in natura. Twenty five compounds were identified in the crude ethanol extract of the samples 1 and 2. The major compounds belong to the class of flavonols, flavanas, isoflavones and prenylated benzophenones pterocarpans, demonstrating the usefulness of the technique of LC - Orbitrap - FTMS. The chemical profile of samples 1 and 2 of propolis are characterized by the presence of benzophenone. In propolis in natura, ethanol extract, hexane fraction, ethyl acetate and hydroalcoholic, we were able to identify and quantify 8 substances, 1 phenolic acid and 7 flavonoids. In the ethyl acetate fraction of sample 1 the isoflavone formononetin was isolated. It is observed that during the fractionation, starting from propolis in natura crushed to formononetin isolated substance, there was an increase in fungistatic power. It is observed that with fractionation, starting from propolis in natura to formononetin isolated substance, there was an increase in fungistatic power. Regarding antioxidant activity against DPPH radical, it was observed that all samples showed antioxidant power, with the exception of isolated formononetin. In descending order, in relation to antioxidant power, the most nonpolar samples had higher activity than polar samples. Both propolis in natura as crude ethanol extract showed strong activity against Leishmania promastigotes of L. amazonensis. The antileukemic activity on the cell line HL- 60 also increased with fractionation. |
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Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, BrasilFracionamento biomonitoradoPrópolis vermelhaPerfil palinológicoBioassay-guided fractionationRed propolisPalynological profileFormononetinCIENCIAS BIOLOGICAS::FARMACOLOGIAPropolis is an heterogeneous mixture of substances, produced by melliferous bees from plants in close proximity with the bee´s hives. It is usually classified according to its botanical origin and chemical composition that vary according to geographical origin. Studies have shown that red propolis have physicochemical and biological properties that are diverse from other propolis types. There is a scarcety of studies with Brazilian red propolis, and no study with the red propolis from Igarassu (PE, Brazil), and thus its botanical origin, chemical characterization, and pharmacological activities have not been determined. The aim of this study was to perform a bioassay-guided fractionation of two propolis samples, collected from Igarassu, in order to determine the substances responsible for its antimicrobial, antioxidant, antileukemic and leishmanicidal activities, as well as its palynological profile. The samples of propolis were collected in Igarassu-PE, in natura, in May 2012 in two apiaries. They were submitted to palynological analyses to identify the pollen types and their frequency in the samples. Then, samples in natura were powdered, extracted with 96% ethanol and concentrated to obtain the crude ethanol extract. This extract was dissolved in a mixture of methanol:water (1:1, v/v) and partitioned with hexane and ethyl acetate, obtaining the hexane, ethyl acetate and hydroalcoholic fractions. The acetate fraction was submitted to column chromatography with Sephadex LH20 for isolation of the compounds. The propolis in natura was subjected to preliminary phytochemical screening. The ethanol extract was evaluated for identification of compounds by HPLC-MS. The propolis in natura, ethanol extract, hexane fraction, ethyl acetate and hydroalcoholic were submitted to HPLC analysis for identification and quantification of phenolic compounds. A total of 28 different pollen types belonging to 16 families and 30 genera were found in the samples. The most frequent were: Serjania and Dalbergia (sample 1 April 2011), Fabaceae 1 and Dalbergia (sample 2 April 2011), Stigmaphyllon (sample 1 and 2 May 2012). It was found the presence of flavonoids and steroids in propolis in natura. Twenty five compounds were identified in the crude ethanol extract of the samples 1 and 2. The major compounds belong to the class of flavonols, flavanas, isoflavones and prenylated benzophenones pterocarpans, demonstrating the usefulness of the technique of LC - Orbitrap - FTMS. The chemical profile of samples 1 and 2 of propolis are characterized by the presence of benzophenone. In propolis in natura, ethanol extract, hexane fraction, ethyl acetate and hydroalcoholic, we were able to identify and quantify 8 substances, 1 phenolic acid and 7 flavonoids. In the ethyl acetate fraction of sample 1 the isoflavone formononetin was isolated. It is observed that during the fractionation, starting from propolis in natura crushed to formononetin isolated substance, there was an increase in fungistatic power. It is observed that with fractionation, starting from propolis in natura to formononetin isolated substance, there was an increase in fungistatic power. Regarding antioxidant activity against DPPH radical, it was observed that all samples showed antioxidant power, with the exception of isolated formononetin. In descending order, in relation to antioxidant power, the most nonpolar samples had higher activity than polar samples. Both propolis in natura as crude ethanol extract showed strong activity against Leishmania promastigotes of L. amazonensis. The antileukemic activity on the cell line HL- 60 also increased with fractionation.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA própolis é uma mistura heterogênea de compostos e é produzida por abelhas a partir de vegetais encontrados próximos às colmeias. É classificada de acordo com a origem botânica e a composição química que pode variar entre as regiões. Estudos têm demonstrado que a própolis vermelha possui características físico-químicas e biológicas diferenciadas. No estado de Pernambuco, até o momento são poucos os trabalhos realizados com a própolis vermelha, e não há na literatura estudos com a própolis vermelha de Igarassu PE, com isso sua origem botânica, caracterização química e suas atividades farmacológicas, ainda não estão definidas. O objetivo deste estudo foi realizar um fracionamento biomonitorado de duas amostras de própolis vermelha, coletadas em Igarassu PE, a fim de determinar os compostos responsáveis pelas atividades antimicrobiana, antioxidante, antileucêmica e antileishmania, além de traçar um perfil palinológico. As amostras de própolis vermelha foram coletadas em Igarassu-PE, na forma in natura, em maio de 2012, em dois apiários. Foram submetidas à análise palinológica, para identificação dos grãos de pólen e dos tipos polínicos presentes. Em seguida, as amostras in natura foram trituradas, extraídas em etanol 96% e concentradas obtendo-se o extrato etanólico bruto seco. Esse extrato foi suspenso em uma mistura de metanol-água (1:1) e fracionado por separação líquido-líquido com hexano e acetato de etila, obtendo-se as frações hexânica, acetato de etila e hidroalcoólica. A fração acetato foi refracionada em coluna de Sephadex LH-20 para isolamento dos compostos. A própolis in natura foi submetida ao screening fitoquimico preliminar. O extrato etanólico foi avaliado para identificação dos compostos por CLAE-EM. A própolis in natura, extrato etanólico, frações hexânica, acetato de etila e hidroalcoólica foram submetidas a análise do perfil cromatográfico por CLAE, análise do perfil fenólico, identificação e quantificação de compostos fenólicos e avaliação das atividades farmacológicas. Nas amostras in natura, foram encontrados 28 tipos polínicos, pertencentes a 16 famílias e 30 gêneros botânicos. Os tipos polínicos que apareceram em maior frequência foram: Serjania e Dalbergia (amostra 1- abril de 2011), Fabaceae 1 e Dalbergia (amostra 2-abril de 2011), Stigmaphyllon (amostra 1 e 2-maio de 2012). Verificou-se a presença de flavonoides e esteroides na própolis in natura. Foram identificados 25 compostos no extrato etanólico bruto das amostras 1 e 2 de própolis vermelha, onde as principais substâncias identificadas pertencem à classe dos flavonols, flavanas, isoflavonas, pterocarpanos e benzofenonas preniladas, demonstrando a utilidade da técnica de LC - Orbitrap - FTMS. O perfil químico das amostras 1 e 2 de própolis vermelha são caracterizados pela presença de benzofenonas. Na própolis in natura, extrato etanólico, frações hexânica, acetato de etila e hidroalcoólica, foram identificadas e quantificadas 8 substâncias, sendo 1 ácido fenólico e 7 flavonoides. Na fração acetato de etila da amostra 1, foi isolada a isoflavona formononetina. Observa-se que ao longo do fracionamento, partindo da própolis in natura triturada até a substância isolada formononetina, houve um aumento no poder fungistático. Em relação a atividade antioxidante, medida pelo método de DPPH, observou-se que todas as amostras apresentaram poder antioxidante, com exceção da substância isolada. Em ordem decrescente, em relação ao poder antioxidante, as amostras mais apolares tiveram maior atividade que as amostras polares. Tanto a própolis in natura triturada assim como o extrato etanólico bruto apresentaram forte atividade antileishmania sobre as formas promastigotas de L. amazonensis. A atividade antileucêmica também aumentou ao longo do fracionamento, sobre a linhagem celular HL-60.Universidade Federal da ParaíbaBRFarmacologiaPrograma de Pós-Graduação em Produtos Naturais e Sintéticos BioativosUFPBCunha, Emidio Vasconcelos Leitão dahttp://lattes.cnpq.br/0731580374184115Oliveira, Eduardo de Jesushttp://lattes.cnpq.br/1293090609428232Neves, Michelline Viviane Marques das2015-05-14T13:00:09Z2018-07-21T00:24:49Z2015-03-272018-07-21T00:24:49Z2014-08-25info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfNEVES, Michelline Viviane Marques das. Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil. 2014. 184 f. Tese (Doutorado em Produtos Naturais e Sintéticos Bioativos) - Universidade Federal da Paraíba, João Pessoa, 2014.https://repositorio.ufpb.br/jspui/handle/tede/6840porinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2018-09-06T01:40:45Zoai:repositorio.ufpb.br:tede/6840Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2018-09-06T01:40:45Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil |
title |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil |
spellingShingle |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil Neves, Michelline Viviane Marques das Fracionamento biomonitorado Própolis vermelha Perfil palinológico Bioassay-guided fractionation Red propolis Palynological profile Formononetin CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil |
title_full |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil |
title_fullStr |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil |
title_full_unstemmed |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil |
title_sort |
Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil |
author |
Neves, Michelline Viviane Marques das |
author_facet |
Neves, Michelline Viviane Marques das |
author_role |
author |
dc.contributor.none.fl_str_mv |
Cunha, Emidio Vasconcelos Leitão da http://lattes.cnpq.br/0731580374184115 Oliveira, Eduardo de Jesus http://lattes.cnpq.br/1293090609428232 |
dc.contributor.author.fl_str_mv |
Neves, Michelline Viviane Marques das |
dc.subject.por.fl_str_mv |
Fracionamento biomonitorado Própolis vermelha Perfil palinológico Bioassay-guided fractionation Red propolis Palynological profile Formononetin CIENCIAS BIOLOGICAS::FARMACOLOGIA |
topic |
Fracionamento biomonitorado Própolis vermelha Perfil palinológico Bioassay-guided fractionation Red propolis Palynological profile Formononetin CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
Propolis is an heterogeneous mixture of substances, produced by melliferous bees from plants in close proximity with the bee´s hives. It is usually classified according to its botanical origin and chemical composition that vary according to geographical origin. Studies have shown that red propolis have physicochemical and biological properties that are diverse from other propolis types. There is a scarcety of studies with Brazilian red propolis, and no study with the red propolis from Igarassu (PE, Brazil), and thus its botanical origin, chemical characterization, and pharmacological activities have not been determined. The aim of this study was to perform a bioassay-guided fractionation of two propolis samples, collected from Igarassu, in order to determine the substances responsible for its antimicrobial, antioxidant, antileukemic and leishmanicidal activities, as well as its palynological profile. The samples of propolis were collected in Igarassu-PE, in natura, in May 2012 in two apiaries. They were submitted to palynological analyses to identify the pollen types and their frequency in the samples. Then, samples in natura were powdered, extracted with 96% ethanol and concentrated to obtain the crude ethanol extract. This extract was dissolved in a mixture of methanol:water (1:1, v/v) and partitioned with hexane and ethyl acetate, obtaining the hexane, ethyl acetate and hydroalcoholic fractions. The acetate fraction was submitted to column chromatography with Sephadex LH20 for isolation of the compounds. The propolis in natura was subjected to preliminary phytochemical screening. The ethanol extract was evaluated for identification of compounds by HPLC-MS. The propolis in natura, ethanol extract, hexane fraction, ethyl acetate and hydroalcoholic were submitted to HPLC analysis for identification and quantification of phenolic compounds. A total of 28 different pollen types belonging to 16 families and 30 genera were found in the samples. The most frequent were: Serjania and Dalbergia (sample 1 April 2011), Fabaceae 1 and Dalbergia (sample 2 April 2011), Stigmaphyllon (sample 1 and 2 May 2012). It was found the presence of flavonoids and steroids in propolis in natura. Twenty five compounds were identified in the crude ethanol extract of the samples 1 and 2. The major compounds belong to the class of flavonols, flavanas, isoflavones and prenylated benzophenones pterocarpans, demonstrating the usefulness of the technique of LC - Orbitrap - FTMS. The chemical profile of samples 1 and 2 of propolis are characterized by the presence of benzophenone. In propolis in natura, ethanol extract, hexane fraction, ethyl acetate and hydroalcoholic, we were able to identify and quantify 8 substances, 1 phenolic acid and 7 flavonoids. In the ethyl acetate fraction of sample 1 the isoflavone formononetin was isolated. It is observed that during the fractionation, starting from propolis in natura crushed to formononetin isolated substance, there was an increase in fungistatic power. It is observed that with fractionation, starting from propolis in natura to formononetin isolated substance, there was an increase in fungistatic power. Regarding antioxidant activity against DPPH radical, it was observed that all samples showed antioxidant power, with the exception of isolated formononetin. In descending order, in relation to antioxidant power, the most nonpolar samples had higher activity than polar samples. Both propolis in natura as crude ethanol extract showed strong activity against Leishmania promastigotes of L. amazonensis. The antileukemic activity on the cell line HL- 60 also increased with fractionation. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-08-25 2015-05-14T13:00:09Z 2015-03-27 2018-07-21T00:24:49Z 2018-07-21T00:24:49Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
NEVES, Michelline Viviane Marques das. Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil. 2014. 184 f. Tese (Doutorado em Produtos Naturais e Sintéticos Bioativos) - Universidade Federal da Paraíba, João Pessoa, 2014. https://repositorio.ufpb.br/jspui/handle/tede/6840 |
identifier_str_mv |
NEVES, Michelline Viviane Marques das. Fracionamento biomonitorado da própolis vermelha de Igarassu, Pernambuco, Brasil. 2014. 184 f. Tese (Doutorado em Produtos Naturais e Sintéticos Bioativos) - Universidade Federal da Paraíba, João Pessoa, 2014. |
url |
https://repositorio.ufpb.br/jspui/handle/tede/6840 |
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por |
language |
por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal da Paraíba BR Farmacologia Programa de Pós-Graduação em Produtos Naturais e Sintéticos Bioativos UFPB |
publisher.none.fl_str_mv |
Universidade Federal da Paraíba BR Farmacologia Programa de Pós-Graduação em Produtos Naturais e Sintéticos Bioativos UFPB |
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reponame:Biblioteca Digital de Teses e Dissertações da UFPB instname:Universidade Federal da Paraíba (UFPB) instacron:UFPB |
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Universidade Federal da Paraíba (UFPB) |
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UFPB |
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UFPB |
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Biblioteca Digital de Teses e Dissertações da UFPB |
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Biblioteca Digital de Teses e Dissertações da UFPB |
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Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB) |
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diretoria@ufpb.br|| diretoria@ufpb.br |
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