title: Quantitation of flupirtine and its active acetylated metabolite by reversed-phase high-performance liquid chromatography using fluorometric detection.
abstract: A simple, selective and sensitive procedure is described for the quantitation of flupirtine maleate (FLU) and its active acetylated metabolite (Met. 1) in plasma and urine. Using a 0.5-ml sample, a sensitivity of 10 ng/ml is easily achieved with a reversed-phase octadecylsilane (C18) column, and a high-performance liquid chromatographic system with fluorescence detection. Quantitation from plasma involves addition of an internal standard, protein precipitation with acetonitrile and a sample concentrating step, while for urinalysis the samples are taken through a single extraction with methylene chloride. Analytical recoveries of FLU and Met. 1 from plasma averaged greater than or equal to 95%, while from urine only 60 and 50%, respectively, could be recovered. The overall, inter- and intra-day variability for both FLU and Met. 1 averaged 6, 5 and 3%, in plasma, respectively. Standard calibration plots in plasma were linear (r greater than or equal to 0.99) for FLU (range: 0.01-10.0 micrograms/ml) and Met. 1 (range: 0.5-25 micrograms/ml) over the extended range. A slightly modified elution system was employed for quantitation of FLU and Met. 1 in urine.
mesh_terms: D000107:Acetylation; D000631:Aminopyridines / Q000032:analysis* / Q000097:blood; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: C040849:2-amino-3-acetylamino-6-(4-fluorobenzylamino)pyridine; D000631:Aminopyridines; C034161:flupirtine