Abstract: HPLC was applied to the simultaneous determination of 5-hydroxymethylfurfural and ecdysterone in granules of radix achyranthis bientatae for making up prescriptions. SHIMADZU shim-pack VP-ODS column (4.6 mm×250 mm, 4.6 μm) was used for separation. Mixtures containing acetonitrile and water mixed in different ratio were used as the mobile phase in gradient elution. UV-detection at the wavelengths of 279 nm (for 5-hydroxymethylfurfural) and 245 nm (for ecdysterone) was adopted in the determination. Linear relationships between values of peak area and mass concentration of 5-hydroxymethylfurfural and ecdysterone were obtained in the ranges of 0.390-100 mg·L-1 and 3.125-100 mg·L-1, with detection limits (3S/N) of 1.95 ng and 2.01 ng respectively. Results of recovery and precision found were: values of average recovery of 99.7% for 5-hydroxymethylfurfural and 99.1% for ecdysteronen with values of RSD′s (n=5) 0.80% and 0.67% respectively.