OBJECTIVE： To establish a method for the determination of 2（5H）-Furanone in Levocanitine injection. METHODS： HPLC was performed on the column of APS-2 HYPERSIL with mobile phase of 0.05 mol/L potassium dihydrogen phosphate buffer solution （pH4.7）-acetonitrile （35 ∶ 65，V/V） at a flow rate of 1.0 ml/min， the detection wavelength was 205 nm， column temperature was 30 ℃， and injection volume was 20 μl. RESULTS： The linear range of 2（5H）-Furanone was 0.001 974-0.987 mg/ml； the limit of quantitation was 4.0 ng， the limit of detection was 1.2 ng； RSDs of precision， stability and reproducibility tests were lower than 2％； recovery was 97.04％-103.36％（RSD＝0.24％，n＝9）. CONCLUSIONS： The method is simple， accurate， sensitive and reproducible， and suitable for the content determination of 2（5H）-Furanone in Levocanitine injection.