Quantitation of Amphotericin B with Use of High-Pressure Liquid Chromatography

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Abstract
A chemical method for determination of concentrations of amphotericin B in serum and cerebrospinal fluid (CSF) is described. After extraction with methanol, the antibiotic was separated by reverse-phase, high-pressure liquid .chromatography and quantitated by absorption at 405 nm. The lower limit of detection of this assay was 0.02 JLg/ml. Relative standard deviations of <3.6% were noted for multiple determinations of sera containing 0.20 and 1.00 μg of amphotericin B/m!. No interfering peaks were found in extracts of serum or CSF from normal humans or in extracts of serum from patients treated with other drugs and antimicrobial agents, including 5-fluorocytosine. Comparison of the method with microbiological assays showed correlation coefficients of 0.90 and 0.83 for serum and CSF determinations, respectively. This chemical assay is very rapid (<30 min), sensitive. accurate, and specific and appears to be suitable for routine clinical use.

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