Abstract: This study employed an ultrasound-assisted deep eutectic solvent (DES) method to systematically optimize the extraction of polysaccharides from Plukenetia volubilis seeds, for promoting the high-value utilization of this resource. After purifying the crude polysaccharides by cellulose column chromatography, a homogeneous fraction (designated PVP-2) was obtained and systematically analyzed for its structural characteristics and in vitro hypoglycemic activity. The results indicated that choline chloride-urea system (molar ratio 1:2) was the optimal DES. The polysaccharide yield reached 13.67%±0.76% under the optimized conditions: Liquid-to-solid ratio of 50:1 mL/g, water content of 40%, ultrasonic temperature of 62 ℃, and ultrasonic time of 71 min. Structural characterization demonstrated that PVP-2 was an acidic heteropolysaccharide with typical polysaccharide absorption peaks and a monomer composition of rhamnose (8.76%), arabinose (16.45%), xylose (12.34%), galacturonic acid (38.42%), glucose (5.68%), and galactose (18.35%) in molar percentages. The weight-average molecular weight (Mw) was determined to be 9.453×10⁵ Da, the number-average molecular weight (Mn) was 5.703×10⁵ Da, and the molecular weight distribution (Mw/Mn) was 1.658. In vitro hypoglycemic activity assays demonstrated that PVP-2 exhibited a concentration-dependent inhibitory effect on both α-glucosidase and α-amylase, with IC₅₀ values of 0.51±0.03 mg/mL and 0.81±0.06 mg/mL, respectively, indicating its promising potential for hypoglycemic applications. This study establishes an efficient extraction method for polysaccharides from Plukenetia volubilis seeds and offers a theoretical foundation for developing functional foods with auxiliary hypoglycemic functions.