Journal of Chromatography & Separation Techniques
Open Access

Abstract

A Simple Bioanalytical Method for the Quantification of Levetiracetam in Human Plasma and Saliva

Karas K, Kuczynska J, Sienkiewicz-Jarosz H, Bienkowski P and Mierzejewski P

A novel, sensitive and selective ultra-high-performance liquid chromatography - coupled to electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method was developed and validated for the quantification of levetiracetam (LEV), a broad-spectrum antiepileptic drug, in human plasma and saliva. A simple protein precipitation method with acetonitrile as precipitating solvent was used to extract LEV from human plasma and saliva. LEV and an internal standard (fluconazole, IS) were separated on a Kinetex analytical column (5 μm C18 100A, 100 × 2.1 mm) using an isocratic solvent system consisting of methanol, water and 100% formic acid at a ratio of 97:3:0.25 (v/v/v) and a flow rate of 0.2 ml/min over 2 min run time. Detection was performed using target ions of [M+H]+ at m/z 171.0 for LEV and m/z 307.0 for IS in selective ion mode. The method was validated in the calibration range of 1.0-50.0 μg/mL for plasma and 0.5-30.0 μg/mL for saliva. Thus the present HPLC-MS/MS method for determination of LEV in human plasma and saliva, is highly sensitive, rapid with a short run-time of 2 min, can be suitable for high sample throughput. The developed method required minimal sample preparation and less plasma sample volume compared to earlier published LC-MS/MS methods. The validation parameters were found to be well within the acceptance limit.