A sensitive and specific HPLC method was developed and validated for the simultaneous determination of vincamine and its potential degradant (metabolite), vincaminic acid. Chromatographic separation was achieved on a Spheri-5 RP-C8 (5 μm) (220 × 4.6 mm i.d.) column using a mobile phase composed of acetonitrile and 0.05 M sodium acetate, pH 4.0 (30 : 70, v/v) at a flow rate of 1 mL/min. The UV detector was set at 270 nm and the quantitation of the analytes was based on the peak areas. The method was
proven to be accurate and precise with linearity ranges of 0.1 - 50 and 0.4 - 50 μg/mL for vincamine and vincaminic acid, respectively. The limits of detection were 0.03 and 0.08 μg/mL for vincamine and vincaminic acid, respectively. The method was applied to serve two goals. First; stability-indicating assay of vincamine in its pharmaceutical formulations, in addition, the determination of vincaminic acid down to a level of 0.07% in presence of excess of the parent drug. Second; drug monitoring of vincamine and its main metabolite, vincaminic acid in human plasma/urine samples taken from a healthy volunteer treated with 60 mg oral dose of vincamine. The accuracy of the method was satisfactory (recovery > 97%). The overall standard deviation ranged from 1.4 to 2.3%.