Polysaccharides from Exocarpium Citri Grandis: Graded Ethanol Precipitation, Structural Characterization, Inhibition of α-Glucosidase Activity, Anti-Oxidation, and Anti-Glycation Potentials

The endocarp of Exocarpium Citri Grandis (ECG) is abundant in various bioactive components, such as polysaccharides: however, there are few studies on them. Thus, it is highly necessary to carry out further research on the structural characterization and biological activities of ECG polysaccharides (EPs), which are important bioactive substances. In this study, water-extracted EPs were precipitated by ethanol with final concentrations of 50%, 70%, and 90% (v/v), respectively. Three crude polysaccharides (EP50, EP70, and EP90) were fractioned successively. The three polysaccharide fractions were structurally elucidated and were investigated in vitro for their biological activities related to glucose metabolism containing inhibitory effects on &alpha:-glucosidase and non-enzymatic glycosylation and their antioxidant capacities. The main results are summarized as follows: (1) Gradient ethanol precipitation and physicochemical properties of EPs: The yields of EP50, EP70, and EP90 were 11.18%, 0.57%, and 0.18%, respectively. The total sugar contents were 40.01%, 52.61%, and 53.46%, and the uronic acid contents were 30.25%, 18.11%, and 8.17%, respectively. In addition, the three fractions had the same composition of monosaccharides, including rhamnose, arabinose, galactose, glucose, xylose, mannose, galacturonic acid, and glucuronic acid, with differences in the content of neutral and acidic monosaccharides. They all may be branched polymers and spherical conformation, and they were acidic polysaccharides containing esterified and non-esterified uronic acids, pyranose-form sugars, and glycosidic linkages of &alpha:-configuration and &beta:-configuration, with esterification degrees of 32.25%, 28.82%, and 15.58%, respectively. Meanwhile, EP50, EP70, and EP90 were mainly amorphous, and the molecular conformation in solution was a spherical branching polymer without a triple helix structure. The EPs exhibited excellent thermal stability, with their structures remaining stable below 170 &deg:C. (2) In terms of activity research, the results showed that EPs had a good &alpha:-glucosidase inhibitory effect with IC50 values of 1.17 mg/mL, 1.40 mg/mL, and 2.72 mg/mL, respectively, among which EP50 was the best. EP50, EP70, and EP90 displayed antioxidant activity by scavenging DPPH and ABTS radicals as well as oxygen radical absorbance capacity. Among them, EP90 had the strongest antioxidant activity. Furthermore, the EPs showed prominent effects on the inhibitory activity of non-enzymatic glycosylation. In summary, the research on the extraction of polysaccharide from ECG provides a technical reference for the further utilization of ECG resources. This study on antioxidant activity provides theoretical support for their use as a natural antioxidant. As oxidation and glycation are relevant to diabetic complications, the result of this work suggests that EPs may be effective in preventing and treating diabetic complications.