Abstract
Abalone viscera contain sulphated polysaccharides with anti-thrombotic and anti-coagulant activities. In this study, a hydrolysate was prepared from blacklip abalone (Haliotis rubra) viscera using papain and bromelain and fractionated using ion exchange and size exclusion chromatography. Hydrolysates and fractions were investigated for in vitro thrombin inhibition mediated through heparin cofactor II (HCII) as well as anti-coagulant activity in plasma and whole blood. On the basis of sulphated polysaccharide concentration, the hydrolysate inhibited thrombin through HCII with an inhibitor concentration at 50% (IC50) of 16.5 μg/mL compared with 2.1 μg/mL for standard heparin. Fractionation concentrated HCII-mediated thrombin inhibition down to an IC50 of 1.8 μg/mL and improved anti-coagulant activities by significantly delaying clotting time. This study confirmed the presence of anti-thrombotic and anti-coagulant molecules in blacklip abalone viscera and demonstrated that these activities can be enriched with a simple chromatography regime. Blacklip abalone viscera warrant further investigation as a source of nutraceutical or functional food ingredients.
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Acknowledgements
Hafiz Ansar Rasul Suleria was awarded an International Postgraduate Research Scholarship (IPRS), Australia Postgraduate Award (APA) from the Australian Government. This research received funding from the Fisheries Research and Development Corporation (FRDC). We also wish to acknowledge Dr. Gene Wijffels for her invaluable editing of the manuscript.
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Hafiz Ansar Rasul Suleria, Simone A. Osborne and Paul Masci conceived and designed the experiments. Hafiz Ansar Rasul Suleria performed all experiments. Rama Addepalli and Wei Chen provided training and assistance with experiments and analysis. Hafiz Ansar Rasul Suleria and Simone A. Osborne wrote the manuscript. Glenda Gobe supervised and provided advice to Hafiz Ansar Rasul Suleria, contributed reagents and edited the manuscript.
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Suleria, H.A.R., Masci, P., Addepalli, R. et al. In vitro anti-thrombotic and anti-coagulant properties of blacklip abalone (Haliotis rubra) viscera hydrolysate. Anal Bioanal Chem 409, 4195–4205 (2017). https://doi.org/10.1007/s00216-017-0367-x
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DOI: https://doi.org/10.1007/s00216-017-0367-x