Abstract
An HPLC method for determination of amphotericin B (I) in plasma and sputum with piroxicam as internal standard has been developed. Plasma samples containing amphotericin B were collected by solid phase extraction using C2 cartridges. Sputum samples (1 g) were homogenised with acetonitrile and extracted like the plasma and the extract injected onto a 5 μm Nucleosil C18 column. The mobile phase was acetonitrile-water 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). Intra 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.
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