HPLC assay for determination of amphotericin B in biological samples

A fast and selective HPLC method for assaying amphotericin B in biological samples was developed and validated. The chromatographic separation was achieved in less than 12 min on a reverse-phase C-18 column using an acetonitrile-acetic acid-water (52:4.3:43.7, v/v/v) mixture as mobile phase. The flow rate was I mL/min and the effluent was monitored at 406 nm. A linear response over the concentration range 0.1-10.0 mu g/mL was obtained. Intra-day and inter-day RSDs were below 5% for all the sample types. This new HPLC method was applied to assay amphotericin B in plasma and several tissue samples such as kidney, liver, spleen and bone marrow. Application of this method to pharmacokinetic studies in mice and dog is provided. Copyright (c) 2007 John Wiley & Sons, Ltd.