HPLC assay for determination of amphotericin B in biological samples

Abstract 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 1 mL/min and the effluent was monitored at 406 nm. A linear response over the concentration range 0.1–10.0 µg/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 © 2007 John Wiley & Sons, Ltd.