Abstract
Heparin (HEP) is known due to their side effects and the red seaweed Halymenia floresia (Hf) sulfated polysaccharides (SP) are heparinoids. In this study we purified the Hf-SP obtained from an aqueous extract and evaluated their anticoagulant activities. Hf-SP 1 (25°C), Hf-SP 2 (80°C) and Hf-SP 3 (80°C) were sequentially isolated. Hf-SP3 had the highest sulfate content (37.45%). Hf-SP 3 was fractionated by ion exchange chromatography on a DEAE-cellulose column using a NaCl gradient. Fractions were lyophilized and submitted to 0.5% agarose gel electrophoresis. The anticoagulant activity was evaluated by the activated partial thromboplastin time using rabbits plasma and expressed in international units per mg of SP using standard HEP (193 IU mg -1 ). The chromatographic procedure separated into four different SP fractions (F I, F II, F III and F IV) eluted at concentrations of 0.50, 0.75, 1.00 and 1.25 M of NaCl, respectively, reveling among them different marked on charge density, when compared by electrophoresis. F III had the highest anticoagulant activity (10.72 IU mg -1 ), suggesting that the sulfate is important in this process. In conclusion, our results suggest that sequential extractions of Hf-SP are an important biotechnological tool for identification of novel anticoagulants and studies of structural characterization are already in progress.
Highlights
Seaweed sulfated polysaccharides (SP) are widely used in pharmaceutical, cosmetic and food industries
We have previously reported that the red seaweed H. floresia is composed by three crude SP (Hf-SP1, Halymenia floresia (Hf)-SP2 and Hf-SP3)
These polysaccharides showed in vitro anticoagulant activity (APTT test) dependent of the sulfate content
Summary
Seaweed sulfated polysaccharides (SP) are widely used in pharmaceutical, cosmetic and food industries Seaweed SP have been described as anticoagulants for some time, difficulty of the isolation in pure form is still reported (ASSREUY et al, 2008; FARIAS et al, 2000, 2001; LEITE et al, 1998; PUSHPAMALI et al, 2008; RODRIGUES et al, 2009; SILVA et al, 2005) Their heterogeneity and polydispersity limit their structural study (AZEVEDO et al, 2009; FARIAS et al, 2000; MELO et al, 2002; MOURÃO, 2004; PEREIRA et al, 2005; RODRIGUES et al, 2009; YOON et al, 2007; ZHANG et al, 2008).
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