By using single factor and orthogonal experiments, the optimum conditions in the purification of total flavonoids from Indocalamus leaves with polyamide resin were studied. The results showed that the optimal sample condition was 2.60 mg•mL-1 in sample concentration, 20 mL of sample volume, sample flow rate of 1.0 mL•min-1, and the optimum elution condition was eluent of 80%（V/V） ethanol, elution liquid amount of 150 mL, the elution rate of 0.5 mL•min-1, the average desorption rate could run up to 94.54%，the amount and concentration of the eluent were significant factors. flavonoids, flavonols and isoflavones could be identified in color reaction of gradient elution.