Resumo
The reportedly low standard quality of heparin (HEP) for use in cardiac surgeries has led to concern in the Brazilian and international markets. Sulfated polysaccharides (SPs) from seaweeds have been regarded as promising substitutes for HEP. The aim of this study was to sequentially extract total SPs (TSPs) from Caulerpa cupressoides (Chlorophyceae) with papain in 100 mM sodium acetate buffer (pH 5.0) containing 5 mM cysteine and 5 mM EDTA, followed by fractionation by ion-exchange chromatography (DEAE-cellulose), and then evaluate the anticoagulant potential of SP fractions by activated partial thromboplastin time (APTT) using normal human plasma and compare it to standard HEP (193 IU mg-1). The obtained fractions were chemically characterized by chemical composition and agarose gel electrophoresis. The yield was 4.61%, and three fractions of SP (F I, F II and F III) eluted with 0.50, 0.75 and 1.00 M of NaCl, respectively, were observed on chromatography profiles; however, differences in charge densities patterns and degree of resolution among them were revealed by electrophoresis. SPs were capable of modifying APTT only in fractions eluted with 0.75 M of NaCl, whose activities were 23.37 and 25.76 IU mg-1, respectively, and the charge density was prerequisite to activity. Therefore, C. cupressoides is a source of SPs possessing low anticoagulant potential compared to HEP.
The reportedly low standard quality of heparin (HEP) for use in cardiac surgeries has led to concern in the Brazilian and international markets. Sulfated polysaccharides (SPs) from seaweeds have been regarded as promising substitutes for HEP. The aim of this study was to sequentially extract total SPs (TSPs) from Caulerpa cupressoides (Chlorophyceae) with papain in 100 mM sodium acetate buffer (pH 5.0) containing 5 mM cysteine and 5 mM EDTA, followed by fractionation by ion-exchange chromatography (DEAE-cellulose), and then evaluate the anticoagulant potential of SP fractions by activated partial thromboplastin time (APTT) using normal human plasma and compare it to standard HEP (193 IU mg-1). The obtained fractions were chemically characterized by chemical composition and agarose gel electrophoresis. The yield was 4.61%, and three fractions of SP (F I, F II and F III) eluted with 0.50, 0.75 and 1.00 M of NaCl, respectively, were observed on chromatography profiles; however, differences in charge densities patterns and degree of resolution among them were revealed by electrophoresis. SPs were capable of modifying APTT only in fractions eluted with 0.75 M of NaCl, whose activities were 23.37 and 25.76 IU mg-1, respectively, and the charge density was prerequisite to activity. Therefore, C. cupressoides is a source of SPs possessing low anticoagulant potential compared to HEP.
Resumo
The reportedly low standard quality of heparin (HEP) for use in cardiac surgerieshas led to concern in the Brazilian and international markets. Sulfated polysaccharides (SPs) fromseaweeds have been regarded as promising substitutes for HEP. The aim of this study was tosequentially extract total SPs (TSPs) from Caulerpa cupressoides (Chlorophyceae) with papain in 100mM sodium acetate buffer (pH 5.0) containing 5 mM cysteine and 5 mM EDTA, followed byfractionation by ion-exchange chromatography (DEAE-cellulose), and then evaluate theanticoagulant potential of SP fractions by activated partial thromboplastin time (APTT) usingnormal human plasma and compare it to standard HEP (193 IU mg-1). The obtained fractions werechemically characterized by chemical composition and agarose gel electrophoresis. The yield was4.61%, and three fractions of SP (F I, F II and F III) eluted with 0.50, 0.75 and 1.00 M of NaCl,respectively, were observed on chromatography profiles; however, differences in charge densitiespatterns and degree of resolution among them were revealed by electrophoresis. SPs were capableof modifying APTT only in fractions eluted with 0.75 M of NaCl, whose activities were 23.37 and25.76 IU mg-1, respectively, and the charge density was prerequisite to activity. Therefore, C.cupressoides is a source of SPs possessing low anticoagulant potential compared to HEP.(AU)
O baixo padrão dequalidade outrora declarado da heparina (HEP) para o uso em cirurgias cardíacas tem levadopreocupação nos mercados nacional e internacional. Os polissacarídeos sulfatados (PSs) de algasmarinhas têm sido considerados como promissores substitutos para HEP. Objetivou-se a extrairsequencialmente PSs totais (PSTs) da clorofícea Caulerpa cupressoides com papaína em tampãoacetato de sódio 100 mM (pH 5,0) contendo cisteína 5 mM e EDTA 5 mM, fracionar porcromatografia de troca iônica (DEAE-celulose) e avaliar o potencial anticoagulante das frações dePS por meio do tempo de tromboplastina parcial ativada (TTPA), utilizando plasma humanonormal e comparando-se à HEP padrão (193 IU mg-1). As frações obtidas foram caracterizadasquimicamente em composição química e por eletroforese em gel de agarose. O rendimento dePSTs foi 4,61% e os perfis cromatográficos, em DEAE-celulose, indicaram a separação de trêsfrações de PS (F I; F II e F III) eluídas nas concentrações 0,50; 0,75 e 1,00 M de NaCl,respectivamente, revelando, por eletroforese, diferenças em termos de densidade de cargas e graude resolução. Os PSs foram capazes de modificar o TTPA somente nas frações eluídas com 0,75M de NaCl, cujas atividades foram 23,37 e 25,76 IU mg-1, respectivamente, quando a densidadede cargas foi pré-requisito para atividade. Portanto, C. cupressoides é uma fonte de PSs com baixospotenciais anticoagulantes comparados à HEP.(AU)
Assuntos
Lycopodium , Polissacarídeos , Alga MarinhaResumo
The reportedly low standard quality of heparin (HEP) for use in cardiac surgeries has led to concern in the Brazilian and international markets. Sulfated polysaccharides (SPs) from seaweeds have been regarded as promising substitutes for HEP. The aim of this study was to sequentially extract total SPs (TSPs) from Caulerpa cupressoides (Chlorophyceae) with papain in 100 mM sodium acetate buffer (pH 5.0) containing 5 mM cysteine and 5 mM EDTA, followed by fractionation by ion-exchange chromatography (DEAE-cellulose), and then evaluate the anticoagulant potential of SP fractions by activated partial thromboplastin time (APTT) using normal human plasma and compare it to standard HEP (193 IU mg-1). The obtained fractions were chemically characterized by chemical composition and agarose gel electrophoresis. The yield was 4.61%, and three fractions of SP (F I, F II and F III) eluted with 0.50, 0.75 and 1.00 M of NaCl, respectively, were observed on chromatography profiles; however, differences in charge densities patterns and degree of resolution among them were revealed by electrophoresis. SPs were capable of modifying APTT only in fractions eluted with 0.75 M of NaCl, whose activities were 23.37 and 25.76 IU mg-1, respectively, and the charge density was prerequisite to activity. Therefore, C. cupressoides is a source of SPs possessing low anticoagulant potential compared to HEP.
The reportedly low standard quality of heparin (HEP) for use in cardiac surgeries has led to concern in the Brazilian and international markets. Sulfated polysaccharides (SPs) from seaweeds have been regarded as promising substitutes for HEP. The aim of this study was to sequentially extract total SPs (TSPs) from Caulerpa cupressoides (Chlorophyceae) with papain in 100 mM sodium acetate buffer (pH 5.0) containing 5 mM cysteine and 5 mM EDTA, followed by fractionation by ion-exchange chromatography (DEAE-cellulose), and then evaluate the anticoagulant potential of SP fractions by activated partial thromboplastin time (APTT) using normal human plasma and compare it to standard HEP (193 IU mg-1). The obtained fractions were chemically characterized by chemical composition and agarose gel electrophoresis. The yield was 4.61%, and three fractions of SP (F I, F II and F III) eluted with 0.50, 0.75 and 1.00 M of NaCl, respectively, were observed on chromatography profiles; however, differences in charge densities patterns and degree of resolution among them were revealed by electrophoresis. SPs were capable of modifying APTT only in fractions eluted with 0.75 M of NaCl, whose activities were 23.37 and 25.76 IU mg-1, respectively, and the charge density was prerequisite to activity. Therefore, C. cupressoides is a source of SPs possessing low anticoagulant potential compared to HEP.