Determination of vinpocetine and Apovincaminic acid in Human plasma by UPLC-MS/MS and its Application

OBJECTIVE To establish and validate a UPLC-MS/MS for the determination of vinpocetine and apovincaminic acid in human plasma and apply to the determination of clinical samples. METHODS Vinpocetine-d5 and apovincaminic acid-d4 were used as internal standards. Human plasma samples were prepared with acetonitrile to precipitate to protein. The chromatographic column was Waters ACQUITY UPLC^® BEH C₁₈(2.1 mm×50 mm, 1.7 μm), the mobile phase was acetonitrile-water(containing 0.05% formic acid). Gradient elution with a flow rate of 0.35 mL·min^-1. Mass spectrometry conditions: Electrospray ionization source(ESI), positive ion detection mode. RESULTS The linear range were 0.04-20.0 ng·mL^-1(r=0.999 7) for vinpocetine and 0.50-250 ng·mL^-1(r=0.999 6) for apovincaminic acid, and the limit of quantification were 0.01, 0.10 ng·mL^-1, respectively. The recoveries of test sample and internal standard were in the range of 94.81%-105.0%, and the matrix effect were 94.51%-105.0%, RSDs were <5%. The RSDs of intra- and inter-batch precision were all <10%. CONCLUSION The method is specific, rapid, accurate and reproducible. Therefore, it is suitable for the determination of vinpocetine concentration in plasma and can applied to its bioequivalence study.