Chondroitin sulfate/dermatan sulfate (CS/DS) was extracted from Atlantic bluefin tuna (Thunnus thynnus) skin (SGAT) and was purified and characterized. SGAT was characterized by acetate cellulose electrophoresis, FTIR spectroscopy, 13 C NMR spectroscopy and SAX-HPLC. According to the results obtained for specific chondroitinases (ABC and AC) and the SAX-HPLC separation of generated unsaturated repeating disaccharides, the polymer was found to contain a disaccharide monosulfated in positions 6 and 4 of GalNAc and disulfated disaccharides in different percentages. These results were confirmed by 13 C NMR experiments. The average molecular mass was 24.07 kDa, as determined by PAGE analysis. SGAT was evaluated for its in vitro anticoagulant activity via activated partial thromboplastin time, thrombin time and prothrombin time tests. The polymer showed strong inhibitory activity against angiotensin I-converting enzyme (IC 50 ¼ 0.25 mg mL À1 ). Overall, the results suggest that this newly extracted CS/DS can be useful for pharmacological applications. Fig. 3 (A) Anticoagulant activity of the purified CS/DS from Atlantic bluefin tuna skin (SGAT) at different concentrations evaluated by measurement of the activated partial thromboplastin time (aPTT). (B) Anticoagulant activity of SGAT at different concentrations evaluated by measurement of the thrombin time (TT). (C) Anticoagulant activity of SGAT at different concentrations evaluated by measurement of the prothrombin time (PT). All results are expressed as mean AE SD (n ¼ 3) and all measurements were performed in triplicate. 37972 | RSC Adv., 2018, 8, 37965-37975 This journal is
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