Determination of propranolol concentration in small volume of rat plasma by HPLC with fluorometric detection

A simple, rapid and sensitive fluorescence high performance liquid chromatographic method was developed to determine propranolol concentration in the small volume of rat plasma without the solvent extraction step using pronethanol as the internal standard. The analysis was accomplished using a 5 microm CAPCELL PAK analytical cyano column at room temperature and a mobile phase consisted of 1% aqueous acetic acid containing 0.2% triethylamine and acetonitrile (65:35, v/v; pH 3.8). The flow-rate was kept at 0.5 mL/min and column effluent was monitored with a fluorescence detector at an excitation wavelength of 230 nm and an emission wavelength of 340 nm. Retention times for pronethalol and propranolol were 8.5 min and 10.5 min, respectively. Linear regressions for the standard curves were linear in the range 2-800 ng/mL, giving correlation coefficients above 0.998. The detection limit was 1.34 ng/mL. No analytical interference was observed from endogenous components in rat plasma. This simple and sensitive assay method was feasibly applied to the pharmacokinetic study of propranolol after intravenous administration of 2 mg/kg of propranolol to normal and carbon tetrachloride-induced liver cirrhotic rats.