Development of an HPLC method for the determination of mexiletine in human plasma and urine by solid-phase extraction.
A sensitive and specific high-performance liquid chromatography (HPLC) method has been developed and validated for the quantification of mexiletine (MEX) in human plasma and urine. It uses solid-phase extraction (SPE) followed by an automated reversed-phase HPLC with a pre-column derivatization with 4-chloro-7-nitrobenzofurazan (NBD-CI) and UV-vis Absorbance detection. The process was set as: the UV-vis Absorbance wavelength was set at 458nm. Chromatographic separation was performed on a Phenomenex-C(18) Column (Aqua, 150mmx4.6mm i.d. with 5mum particle size) with the mobile phase consisting of acetonitrile and water (80:20, v/v), and the flow rate was set at 1.0mLmin(-1). Calibration of the overall analytical procedure gave a linear signal (r>0.9998) over a MEX concentration range of 0.2-2.0mugmL(-1) in human plasma and urine. The detection limit in plasma and urine was 0.1mugmL(-1). Intra- and inter-day precision of the assay at three concentrations within this range were 0.31-2.50%. The high specificity and sensitivity have been achieved by this fast method (total run-time <6min). The method has been successfully validated in human plasma and urine and it has been shown to be precise, accurate and reliable.
Ulu
Sevgi Tatar
ST
Department of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, Istanbul 34452, Turkey.