Objective:The refining parameters of isoflavones from Puerariae flos were optimized by using HPD-100 macroporous adsorption resin, and its in vitro lipid-lowering effects were evaluated. Methods:The optimized refine parameters of isoflavones from Puerariae flos were studied by macroporous adsorption resin, and the lipid-lowering effects of isoflavones from Puerariae flos were evaluated by pancreatic lipase enzyme activity, pancreatic cholesterol esterase enzyme activity and micellar cholesterol solubility. Results:Adsorption parameters: Puerariae flos isoflavones were adsorbed with concentration of 100 mg/mL, pH 6.0 and upper column volume of 20 mL. Desorption parameters: the adsorption resins were washed by 70% ethanol with pH 5.0 at flow rate of 2.0 mL/min. The purity of refined Puerariae flos isoflavones were 81.47%. The inhibition ratio on pancreatic lipase enzyme, pancreatic cholesterol esterase enzyme and micellar cholesterol solubility were 40.73%, 52.32% and 46.17%, respectively, and these were increased by 26.65%, 47.46% and 37.53% respectively, compared with not refining. Conclusion:Puerariae flos isoflavones showed lipid-lowering effects by inhibiting pancreatic lipase enzyme activity, pancreatic cholesterol esterase enzyme activity and micellar cholesterol solubility.