An HPLC method was established for the amount determination of fluconazole in fluconazole injection.The anal. was performed on a C18 column (150 mm × 4.6 mm, 5 μm).The mobile phase was composed of phosphate buffer solution (pH = 7.0)-methanol (55:45) with the flow rate of 1.0 mL/min.The detection wavelength was 260 nm.The standard curve was linear in the range of 0.21-2.13 μg for fluconazole (r = 0.9998).The recovery rate was 99.74% with RSD = 0.90 (n = 9).The method was accurate, sensitive, and suitable for the amount determination of fluconazole for injection.