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Determination of Uridine in Human Plasma by HPLC and its Application in Citicoline Sodium Pharmacokinetics and Bioequivalence Studies

Keguang Chen, Xiaoyan Liu, Chunmin Wei, Guiyan Yuan, Rui Zhang, Rong Li, Benjie Wang and Ruichen Guo

A simple and rapid high performance liquid chromatography (HPLC) method was developed and validated to determine uridine, metabolite of citicoline, in human plasma, and applied to the pharmacokinetics and bioequivalence studies of citicoline sodium tablet and capsule. Uridine and amoxicillin (used as internal standard, IS) were extracted from plasma by simple protein precipitation with perchloric acid. The separation was performed on a Phenomenex kinetex C18 (100 ×4.6 mm, 2.6μm) column, with a mobile phase of 0.05 M phosphate buffer (Potassium dihydrogen phosphate, adjusted pH to 3.5 by phosphate) - Methanol (98:2, V/V), and delivered at the flow rate of 0.8 mL/min. The calibration curve was linear over uridine concentration range of 0.05-2μg/mL (r>0.99), The main pharmacokinetic parameters of citicoline sodium tablet and capsule were as follows, t 1/2 (4.393±2.526) and (4.857±4.503) h, T max (3.354±1.118) and (3.688±1.082) h, Cmax (1.956±0.402) and (2.070±0.619) μg/mL, AUC0-12 (12.774±3.222) and (13.992±3.953) μg/mL*h, AUC0~∞ (16.015±5.647) and (17.198±6.672) μg/mL*h, respectively. The two-one side t-test analysis showed that the confidential intervals of Cmax, AUC0-12, AUC0~∞ of citicoline sodium tablet and capsule were (89.5%~102.1%), (85.3%~97.5%), (85.3%~102.9%), respectively. The relative bioavailability of citicoline sodium tablets was 92.7%. The tablet and capsule were bioequivalence.