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

A robust, fully automated assay procedure for the determination of rosiglitazone (I, BRL-49653) in human plasma has been developed. Plasma concentrations of I were determined using automated sequential trace enrichment of dialysates (ASTED) coupled to reversed-phase high-performance liquid chromatography. Sequential automated dialysis of human plasma samples was followed by concentration of the dialysate by trace enrichment on a C-18 cartridge. Drug and internal standard, SE-204882 (II) were eluted from the trace enrichment 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 compounds were detected by fluorescence detection, using an excitation wavelength of 247 nm, and emission wavelength of 367 nn. The lower limit of quantitation 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 determination were better than 10% across the calibration range. There was no evidence of instability of I in human plasma following three complete freeze-thaw cycles and samples can be safely stored for at least 7 months at -20 degreesC. This method has been successfully utilised to provide pharmacokinetic data throughout the clinical development of rosiglitazone. (C) 2001 Elsevier Science B.V. All rights reserved.