Automated high-performance liquid chromatography method for the determination of rosiglitazone in human plasma

A robust, fully automated assay procedure for the determination of rosiglit azone (I, BRL-49653) in human plasma has been developed. Plasma concentrati ons of I were determined using automated sequential trace enrichment of dia lysates (ASTED) coupled to reversed-phase high-performance liquid chromatog raphy. Sequential automated dialysis of human plasma samples was followed b y concentration of the dialysate by trace enrichment on a C-18 cartridge. D rug and internal standard, SE-204882 (II) were eluted from the trace enrich ment cartridge by mobile phase (0.01 M ammonium acetate, pH 8-acetonitrile, 65:35, v/v) onto the HPLC column (a Novapak C-18, 4 mum, 100 x 5 mm radial compression cartridge) protected by a Guard-Pak C-18 cartridge. The compou nds were detected by fluorescence detection, using an excitation wavelength of 247 nm, and emission wavelength of 367 nn. The lower limit of quantitat ion of the method was 3 ng/ml (200 mul aliquot) with linearity demonstrated up to 100 ng/ml. Within- and between-run precision and accuracy of determi nation were better than 10% across the calibration range. There was no evid ence of instability of I in human plasma following three complete freeze-th aw cycles and samples can be safely stored for at least 7 months at -20 deg reesC. This method has been successfully utilised to provide pharmacokineti c data throughout the clinical development of rosiglitazone. (C) 2001 Elsev ier Science B.V. All rights reserved.