Automated high-performance liquid chromatography method for the determination ofrosiglitazone inhuman plasma

摘要

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 C18 cartridge. Drug and internal standard, SB-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 C18, 4 g~am, 100X5 mm radial compression cartridge) protected by a Guard-Pak C11~ cartridge. The compounds were detected by fluorescence detection, using an excitation wavelength of 247 nm, and emission wavelength of 367 rim. The lower limit of quantitation of the method was 3 ng/ml (200 jil 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 — 200(2. This method has been successfully utilised to provide pharmacokinetic data throughout the clinical development of rosiglitazone.