Determination of amphotericin B in human plasma using solid-phase extraction and high-performance liquid chromatography

• Published in: Journal of pharmaceutical and biomedical analysis 2001-04, Vol.25 (1), p.53-64
• Main Authors: Eldem, T, Arican-Cellat, N
• Format: Article
• Language: English
• Subjects: 1-Amino-4-nitronaphthalene; Amphotericin B; Amphotericin B - blood; Analysis; Antifungal Agents - blood; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; General pharmacology; High pressure liquid chromatography; Humans; Medical sciences; Pharmacology. Drug treatments; Plasma amphotericin B concentration; Quality Control; Reference Standards; Reproducibility of Results; Solid phase extraction; Validation
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(00)00499-4

A rapid and selective HPLC method is described and validated for measuring amphotericin B (AB) in plasma. The procedure involves the solid phase extraction of AB from plasma by incorporating 1-amino-4-nitronaphthalene as an internal standard during the last elution step in extraction followed by HPLC analysis with UV detection at 407 nm. The chromatographic separation is achieved in less than 10 min on a reversed-phase C-18 column using acetonitrile-disodium edetate (20 mM) (45:55, v/v) at pH 5.0 as eluent. A linear response over the concentration range of 0.0100–2.00 μg ml −1 is obtained having a detection limit of 0.00500 μg ml −1 for AB. The mean extraction recovery is found to be 98.1±1.1% ( n=15). The within-day and day-to-day R.S.D. were less than 2% ( n=15) and 6.54% ( n=45) respectively. This method is applied for quantifying AB trough levels in the plasma of cancer patients who have been on antifungal therapy with AmBisome®. It can further be applied either for AB therapeutic monitoring or single/multiple pharmacokinetic analysis of AB in plasma.