Abstract: To investigate the desirable purification process of ponkan Chenpi flavonoids (PCf) and their in vitro antioxidant and hypoglycemic activities, this study extracted PCf using ultrasound-assisted solvent extraction. Four types of macroporous resins were evaluated for their static adsorption-desorption efficiency toward PCf, and HPD-500 resin was identified as the optimal carrier. Single-factor experiments were subsequently conducted to optimize the purification parameters. The antioxidant activity of PCf (before and after purification) were assessed using DPPH radical scavenging rate, ABTS⁺ radical scavenging rate, and ferric ion reducing capacity. The α-glucosidase inhibitory activity was evaluated. Results demonstrated that HPD-500 resin achieved the highest purification efficiency for PCf under optimized conditions: 2 of pH, 80% ethanol concentration, sample loading flow rate of 4 mL/min, elution flow rate of 3 mL/min, and a sample-to-eluent volume ratio of 1:1.6. Post-purification, the reducing sugar content of PCf was significantly reduced, while the total flavonoid content increased (132.2±9.62 mg RE/g DW). Purified PCf exhibited stronger antioxidant and hypoglycemic activities, with 93.75%±0.82% of ABTS⁺ radical scavenging rate reaching, 77.42%±0.91% of DPPH radical scavenging rate, double ferric ion reducing capacity, and 66.84%±0.67% of α-glucosidase inhibitory rate. In summary, HPD-500 resin is suitable for the preliminary separation and purification of ponkan Chenpi flavonoids, and the purified substance demonstrates significant antioxidant activity and inhibitory effect of α-glucosidase, providing a valuable reference for the further development of ponkan Chenpi-based functional products.