Determination of metoprolol in human plasma and urine by high-performance liquid chromatography with fluorescence...

A simple, specific and sensitive HPLC method has been developed for the determination of metoprolol in human plasma and urine. Separation of metoprolol and atenolol (internal standard) was achieved on an Ace C(18) column (5 microm, 250 mm x 4.6 mm id) using fluorescence detection with lambda(ex)=276 nm and lambda(em)=296 nm. The mobile phase consists of methanol-water (50:50, v/v) containing 0.1% TFA. The analysis was performed in less than 10 min with a flow rate of 1 mL/min. The assay was linear over the concentration range of 3-200 and 5-300 ng/mL for plasma and urine, respectively. The LOD were 1.0 and 1.5 ng/mL for plasma and urine, respectively. The LOQ were 3.0 and 5.0 ng/mL for plasma and urine, respectively. The extraction recoveries were found to be 95.6 +/- 1.53 and 96.4 +/- 1.75% for plasma and urine, respectively. Also, the method was successfully applied to three patients with hypertension who had been given an oral tablet of 100 mg metoprolol.