OBJECTIVE: To prepare Puerarin polymeric micelles and establish a method to determine its entrapment efficiency. METHODS: Puerarin polymeric micelles were prepared by film dispersion method. The polymeric micelles and free drug were separated by centrifugal-millipore filter filtration method. The entrapment efficiency of puerarin polymeric micelles was determined by HPLC. Diamonsil C18(2) column was used with 1% citric acid solution-methanol (65 ∶ 35) at the flow rate of 1 ml/min. The detection wavelength was set at 250 nm, and column temperature was room temperature. RESULTS: The prepared polymeric micelles were spherical and spherical-like in shape with a mean particle size of 54.12 nm, polydispersity index of 0.122, Zeta potential of -13.60 mV; the linear range of puerarin was 2-10 μg/ml (R2=0.999 4) with average recovery rate of 99.2% (RSD=0.9%, n=3). The recovery rate of free drug was 95.3% (RSD=1.7%, n=3). The mean entrapment efficiency and drug-loading amount of puerarin were(35.5±2.12)% and (0.3±0.07)%, respectively (n=3). CONCLUSIONS: Film dispersion method is suitable for the preparation of Puerarin polymeric micelles. Established method is convenient, accurate and reliable for the content and entrapment efficiency determination of Puerarin polymeric micelles.