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Determination ofmetoprolol in rabbit plasma by high-performance liquid chromatography with fluorescence detection

BilalYilmaz

A simple, specific and sensitive high-performance liquid chromatography (HPLC) method has been developed for the determination of metoprolol in rabbit plasma. Separation of metoprolol and atenolol (internal standard) was achieved on a Ace C18 column (5m, 2504.6 mm i.d.) using fluorescence detection with ex=276 nmand em=296 nm. The mobile phase consisted of methanol-water (50:50, v/v) containing 0.1% trifluoroacetic acid (TFA). The analysis was performed in less than 5.0 min with a flow rate of 1ml/min. Calibration curvewas linear over the concentration range 3-200ng/ ml. Intra- and inter-day precision values for metoprolol in rabbit plasma were less than 5.0, and accuracy (relative error) was better than 4.8%. The analytical recovery ofmetoprolol fromrabbit plasma averaged out to 95.7%. The limits of detection (LOD) and quantification (LOQ) ofmetoprololwere 1.0 and 3.0ng/ml, respectively. Also the developed and validated HPLC method was successfully applied to a pharmacokinetic study of metoprolol in New Zealand white rabbits.