Objective To develop a high-performance liquid chromatography (HPLC) method for simultaneous determination of five flavonoids (chamaechromone, neochamaejasmin A, chamaejasmine, isochamaejasmin and 7-methoxylneochaejasmin A) in the extraction of Stellera chamaejasme L.. Methods An HPLC-diode array detector (DAD) method was established. The HPLC-DAD condition was as follows:chromatographic column was Agilent Eclipse Plus C₁₈ (3.0 mm×100 mm, 3.5 μm); mobile phase A was 0.1% formic acid-water solution; mobile phase B was acetonitrile; gradient of acetonitrile was 20% (0-6 min), 20%-35% (6-10 min) and 35% (10-50 min); flow rate was 0.4 mL/min; equilibration time was 10 min; acquisition time was 50 min; column temperature was 25℃; DAD detection wavelength was 290 nm; and sample injection volume was 5 μL. Results Chamaechromone, neochamaejasmin A, chamaejasmine, isochamaejasmin and 7-methoxylneochaejasmin A were successfully separated using this method, with good linear relationship between 4.680-468.0, 2.016-201.6, 3.784-378.4, 5.520-552.0 and 0.974-97.40 μg/mL, respectively. The precision, stability, repeatability and recovery of the five flavonoids were good with this method. Conclusion HPLC-DAD has good stability and repeatability, and can be used to determine the contents of five flavonoids (chamaechromone, neochamaejasmin A, chamaejasmine, isochamaejasmin and 7-methoxylneochaejasmin A) in Stellera chamaejasme L..