Abstract: To establish a HPLC method for simultaneous determination of lobetyolin and syringin in Codonopsis Radix from different places. The HPLC system consisting of Thermo scientific C₁₈ Column(4.6 mm×250 mm,5 μm) and a gradient elution system of acetonitrile-water as the mobile phase was adopted. The flow rate was set at 1.0 m L/min,the detection wavelength was 266 nm,and the column temperature was 25 ℃. The results indicated that the chromatographic peaks of lobetyolin and syringin could be separated well. The linear ranges for lobetyolin and syringin were 10 ~200 μg/m L and 0.5 ~10 μg/m L,respectively. The average recoveries were100.65% and 98.78%,respectively. The lobetyolin content and syringin content in Codonopsis Radix from ten places was ranged from 0.1963 ~1.4532 mg/g and 0.0051 ~0.0951 mg/g,respectively. The simple,quick,reproducible and reliable method could be used for determination of two main components in Codonopsis Radix.