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Determination of amphotericin B in human serum by liquid chromatography
R Lopez-Galera1, L Pou-Clave, C Pascual-Mostaza
1Biochemistry Service, Hospital General Universitario Vall d'Hebron, Barcelona, Spain.
Abstract:
A rapid reversed-phase high-performance liquid chromatographic method with a 30-mm long column is described for assaying amphotericin B in serum. After deproteinization of serum samples with methanol, the supernatant was injected onto a reversed-phase C18 column, using 2.5 mM Na2EDTA-acetonitrile (70:30, v/v) as the mobile phase. Amphotericin B was eluted at 1.5 min. Calibration plot of the peak area against concentration was linear from 0.05 to 25 micrograms/ml (C.V. of 3%). Within-day and day-to-day imprecision (C.V.) ranged between 1.33% and 3.61%. The application was evaluated in 55 serum samples from patients treated with amphotericin B.