Mise au point et validation d'une méthode de dosage de l'amphotéricine B par chromatographie liquide à haute performance

Amphotericin B is an effective antifungal drug used in the treatment of systemic mycosis, but it has a narrow therapeutic interval and an important renal toxicity. The aim of this study is to monitor and valid a new method for Amphotericin B quantitation in plasma using HPLC with UV/visible detection. A liquid-liquid extraction was done; thereafter, a chromatographic separation was achieved using an RP 18 column (125 mm). The mobile phase is composed of water, acetonitrile and acetic acid (51/44/5) (v/v/v), pH 3. Piroxicam is used as an internal standard. This technique is linear over the range 0,125 μg/mL to 8 μg/mL (r = 0,99). Detection and quantification limits were 0.044 μg/mL and 0.111 μg/mL, respectively. Within-day coefficient of variation (1,98 to 4,66 %) and day-to-day coefficient of variation (2,69 to 4,95 %) were determined at three different concentrations. Under these conditions, each analysis requires no longer than 10 min.