Abstract
The objective of this study was to determine the effect of hydrothermal-assisted extraction (HAE) and a multi-step purification process on the molecular characteristics and antioxidant activity of extracts of laminarin from the brown seaweed Laminaria hyperborea. The HAE with optimised extraction conditions (time - 30 min; temperature - 99.3 °C; sample to solvent ratio - 1:21.3 (w/v)) resulted in 368 mg/g laminarin in the Crude Extract. Purification using solvents, molecular weight cut-off filter (MWCO; 10 kDa) and solid-phase extraction (SPE) improved the concentration to 542.6 mg/g, 906.6 mg/g and 862.5 mg/g on dry weight extract basis, respectively (p < 0.05). The crude HAE extract had greater antioxidant activity, as determined by 1,1-diphenyl-2-picryl-hydrazyl radical scavenging and ferric reducing antioxidant power assays, than the purified fractions (p > 0.05). The identification of a chromatographic peak at 17.66 min (retention time) and FT-IR (Fourier transform infrared) absorption bands at 1420 cm−1 (carboxyl groups), 2950 cm−1 (C-H stretch) and at 2410 cm−1 (transmitting angle peak) was similar between the purified samples and a laminarin standard that was used as a control, confirming the presence of laminarin. The quadruple time of flight mass spectrometry (Q-ToF-MS) confirmed the molecular weight of purified laminarin was in the range of 5.7 kDa–6.2 kDa. HAE under optimised conditions was an effective method to recover biologically active laminarin from L. hyperborea. A multi-step purification, involving solvents and MWCO filters in a sequence could improve the purity of laminarin, however, purification reduced the antioxidant activity compared to the crude HAE extract.
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