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Abstract
A liquid chromatographic method incorporating column-switching for the separation and determination of chlorthalidone enantiomers in urine is described. Untreated urine samples (50 μL) were directly introduced into a 20 mm x 2.1 mm I. D. precolumn packed with a Hypersil ODS-C18, 30 μm, stationary phase. Polar urinary compounds were removed by flushing the precolumn with 4 mL of water (pumped at a flow rate of 1 mL/min), and the racemic analyte was then transferred to a LiChroCART ChiraDex, 5 μm, 250 mm x 4 mm I. D. column, where the enantiomers were separated by means of a methanol/0.05 M acetate buffer adjusted to pH 4 mobile-phase (40:60, v/v), and quantified at 230 nm. The system shows good linearity and reproducibility in the 0.25 - 5.0 μg/mL, thus covering therapeutic levels of chlorthalidone in urine. The usefulness of the described procedure has been tested by analyzing urine samples obtained after drug administration.