simple and sensitive HPLC method for fluorescence quantitation of doxorubicin in micro-volume plasma: applications to pharmacokinetic studies in rats

ABSTRACT

A validated HPLC method was developed to determine the doxorubicin concentration in a small volume of rat plasma [60 microL] with convenient fluorescence detection. Sample preparation includes a simple one-step liquid-liquid extraction using a minimum amount of organic solvent, with extraction recovery more than 95%. The analysis was accomplished using PerfectSil C18 column maintained at 35 [degree]C and a mobile phase consisted of acetonitrile and water [3268, v/v; pH=2.6]. The flow-rate was kept at 1 mL/min and the column effluent was monitored with a fluorescence detector at an excitation and emission wavelength of 470 and 555 nm, respectively. The detection limit was 5 ng/mL. No analytical interference was observed from endogenous components in the rat plasma. This method was feasibly applied to the pharmacokinetic study of 5 mg/Kg of doxorubicin after the intravenous administration to rats