Automated determination of reboxetine by high-performance liquid chromatography with column-switching and ultraviolet detection

A fully automated method including column-switching and isocratic high-performance liquid chromatography (HPLC) was developed for quantitative analysis of the new antidepressant reboxetine, a noradrenaline reuptake inhibitor. After serum injection into the HPLC system and on-line sample clean-up on a silica C8 (10×4.0 mm I.D.) clean-up column with an eluent consisting of 2.5% acetonitrile in deionized water, the chromatographic separation was performed on an analytical column (Lichrospher CN; 250×4.6 mm I.D.) with an eluent of acetonitrile–aqueous potassium phosphate buffer (0.008 M, pH 6.4) (50:50). The UV detector was set at 273 or 226 nm. The limit of quantification was about 15 ng/ml at 273 nm and about 4 ng/ml at 226 nm. The day-to-day relative standard deviation ranged between 2.7 and 6.7% with recovery rates ≥90%. Linear regression analyses revealed correlation coefficients >0.998. The method can be applied to therapeutic drug monitoring of reboxetine as well as pharmacokinetic studies.