DEVELOPMENT AND VALIDATION OF AN HPLC METHOD FOR DETERMINATION OF AMPHOTERICIN-B IN PLASMA AND SPUTUM INVOLVING SOLID-PHASE EXTRACTION

An HPLC method for determination of amphotericin B (I) in plasma and s putum with piroxicam as internal standard has been developed. Plasma s amples containing amphotericin B were collected by solid phase extract ion using C-2 cartridges. Sputum samples (1 g) were homogenised with a cetonitrile and extracted like the plasma and the extract injected ont o a 5 mu m Nucleosil C-18 column. The mobile phase was acetonitrile-wa ter acetic acid (44:51:5 v/v). Detection was UV absorbance at 407 nm. The calibration graph was linear from 10 to 2000 ng mL(-1) of (I). Int ra and inter-day CV did not exceed 13.4 %. The quantitation limit was 5 ng mL(-1) for plasma and 10 ng mL(-1) for sputum respectively.