Quantification of Levetiracetam in Human Plasma with Precipitation Extraction by Using Fully Validated LC-MS/MS and Its Application to a Bioequivalence Study

A simple, sensitive and selective method for the determination of Levetiracetam by using rapid high-performance liquid chromatography/positive electroscopy ionization tandem mass spectroscopy was developed. The method consists of precipitation extraction with Acetonitrile followed by the analysis of the extracted sample by liquid chromatography-mass spectroscopy (LC-MS/MS) in selective reaction monitoring mode using electrospray ionization mode (ESI). Chromatography was performed on a C₁₈ reverse phase column, Methanol: Acetonitrile: 10mM ammonium acetate (45:45:10) as a mobile phase. The assay exhibited a linear dynamic range of 1 to 40μg/ml for Levetiracetam in human plasma. Stability assessment was also included. A run time of 3.0 min for each sample made it possible to analyse healthy volunteers participating in pharmacokinetics drug-drug interaction studies.