To establish a high performance liquid chromatog. method for the determination of phosphocholine in citicoline. Separation was performed on a C18(4.6 mmx150 mm, 5.0 μm)column with ammonium formate buffer solution(10 mmol/L ammonium formate(with the pH value adjusted to 3.0 with formic acid)-acetonitrile(98:2))as mobile phase and acetonitrile as gradient eluent both at a flow rate of 1.0 mL/min. The column temperature was set at 30°C, the sample volume was 25 μL, the atomization temperature was set at 40°C, the Filter was 10.0 and the acquisition frequency was 10 Hz. Results: Under this method, the detection limit of choline phosphate was 0.2025 μg/mL, and the linear relationship was good in the range of 2.0684-103.4191 μg/mL. After the destructive test, the related substances in citicoline were well separated from phosphorylcholine. The accuracy recovery rate was 102.3%, the RSD was 0.7%. And the chromatog. conditions had slightly changed, which meet the detection requirements. The exptl. operation of this method was convenient, the method was simple, and it was suitable for the determination of phosphocholine in citicoline.