Abstract: This study prepared American ginseng saponins/flavonoids/polysaccharides/soy protein isolate nanoparticles (AG Nps) by the reverse solvent precipitation method. The response surface method was used to optimize the preparation process of AG Nps. The particle size analysis, Zeta potential determination, Fourier transform infrared spectroscopy analysis, differential scanning calorimetry analysis and thermogravimetric analysis of AG Nps were carried out. The in vitro antioxidant, hypolipidemic, and hypoglycemic activities of AG Nps were evaluated. The results indicated that the optimum conditions for the preparation of AG Nps were reverse solvent phase concentration 16 mg/mL, solvent phase concentration 15 mg/mL, volume ratio 39:1. The encapsulation efficiency of AG Nps under these conditions was 90.14%±1.03%, and drug loading was 1.11%±0.07%. The average particle size was 318.60±5.60 nm with uniform distribution, and Zeta potential was −33.05±0.50 mV. The thermostability and antioxidant capacity of nanoparticles improved. The half-inhibitory concentration (IC₅₀) of pancreatic lipase and cholesterol micelles was 7.29 mg/mL and 6.32 mg/mL respectively, indicating that AG Nps has certain inhibitory effects on pancreatic lipase and cholesterol micelles and better cholate binding abilities. The inhibitory activities of α-glucosidase and α-amylase significantly increased (P<0.05), with IC₅₀ values of 3.21 mg/mL and 5.33 mg/mL respectively. In conclusion, AG Nps possess great in vitro hypolipidemic and hypoglycemic activities. These findings provide valuable theoretical and technical insights for the development of functional products of American ginseng.