QUANTITATIVE DETERMINATION OF LEVETIRACETAM IN HUMAN URINE USING HPLC-UV AND ITS IDENTIFICATION BY LC-ESI-MS

Abstract

A rapid method for the quantification of levetiracetam (LV) in human urine using high-performance liquid chromatography combined with UV detection (HPLC–UV) was developed and validated for linearity, repeatability, limit of detection, and limit of quantitation. In addition, a high-performance liquid chromatography combined with electrospray ionization mass spectrometry was described for its identification in human urine. Chromatographic separation was achieved on 250 × 4.6 mm (i.d.) Discovery® (5 µm particle size) reversed-phase C₁₈ analytical column using 0.1% formic acid in water–acetonitrile, in ratio of 85:15, v/v as mobile phase and UV detection at 210 nm for the HPLC–UV method and using 0.1% formic acid in water–acetonitrile, in ratio of 75:25, v/v as mobile phase for LC-ESI-MS method. The HPLC–UV method was successfully applied for establishment of the urinary excretion pattern of LV after oral dose.

Keywords:

• HPLC-UV
• LC-ESI-MS
• levetiracetam
• urinary excretion

Notes

^a Y = a + bC, where C is the concentration of LV in µg mL⁻¹ and Y is the peak area.

^b 95% confidence limit.

^a Mean and S.D. for five determinations.