An HPLC method was established for determination of epirubicin hydrochloride for injection.The chromatog. column was C18 (250 mm×4.6 mm, 5 μm).The mobile phase was acetonitrile-water-methanol-85% phosphoric acid solution (290:540:170:1) containing 0.2% sodium dodecyl sulfate.The flow rate was 1.0 mL/min, and the detection wavelength was 254 nm.The column temperature was 35°C and the sample size was 20 μL.The linear range of epirubicin hydrochloride was 0.0518-0.1554 mg/mL (r=0.9999) and the average recovery was 99.64% (RSD=0.71%).This method was simple, fast, accurate and specific, which could be used for the determination of epirubicin hydrochloride for injection.