Microwave-assisted aqueous two-phase extraction of polysaccharides from Hippophae rhamnoide L.: Modeling, characterization and hypoglycemic activity

Abstract

Natural polysaccharides are concerned for their high biological activity and low toxicity. Two kinds of polysaccharides were extracted from Hippophae rhamnoide L. by microwave-assisted aqueous two-phase system. Under the optimal conditions predicted by RSM model (K₂HPO₄/ethanol (18.93 %/28.29 %), liquid to material ratio 77 mL/g, power 625 W and temperature 61 °C), the yield of total polysaccharides reached 35.91 ± 0.76 %. Moreover, the polysaccharides extraction was well fitted to the Weibull model. After purification by Sepharose-6B, the polysaccharides from top phase (PHTP, purity of 81.44 ± 1.25 %) and bottom phase (PHBP, purity of 88.85 ± 1.40 %) were obtained. GC, FT-IR, methylation and NMR analyses confirmed the backbone of PHTP was composed of a repeated unit →4)-β-D-Glcp-(1 → 2)-α-L-Rhap-(1 → 4)-β-D-Galp-(1 → 4)-α-D-GalAp-(1 → 3)-α-L-Araf-(1 → 3)-α-D-Manp-(1→, while the repeated unit in PHBP was →3)-α-L-Araf-(1 → 2)-α-L-Rhap-(1 → 4)-β-D-Glcp-(1 → 3)-α-D-Manp-(1 → 4)-β-D-Galp-(1 → 4)-α-D-GalAp-(1→. Compared with PHTP (6.46 × 10⁶ g/mol), PHBP with relatively low molecular weight (8.2 × 10⁵ g/mol) exhibited the smaller particle size, better water-solubility, thermal and rheological property, stronger anti-glycosylation and α-amylase inhibitory effects. Moreover, PHTP and PHBP displayed a reversible inhibition on α-amylase in a competitive manner. This study provides a high-efficient and eco-friendly method for polysaccharides extraction, and lays a foundation for sea buckthorn polysaccharides as potential therapeutic agents in preventing and ameliorating diabetes.

Keywords: Characterization; Hypoglycemic activity; Microwave-assisted aqueous two-phase extraction.