Mitoxantrone (DHAQ) is a new anti-cancer agent. The ion-pair HPLC method to determine mitoxantrone hydrochloride injection was proposed. Mitoxantrone can be separated on Zorbax-ODS column with methanol-water-formic acid (450:50:1.5) containing 3.605g SDS as mobile phase. The pH of mobile phase was adjusted to 2.7. The flow rate was 1.0 ml/min and detection was at wavelength of 254 nm, providing a detection limit of 1ng (r/n, 3:1) of DHAQ. DHAQ was determined by peak area measurement with ametantrone as internal standard. The rectilinear calibration graph in the range of 1-10 micrograms of DHAQ was obtained (r = 0.999 2). The average recovery was 99.62 +/- 2.36% (n = 10), and the results were not interfered by the other ingredients in the injection.