title: Improved high-performance liquid chromatographic determination of amphotericin B in human serum and plasma.
abstract: An improved reversed-phase high-performance liquid chromatographic procedure is described for the determination of Amphotericin B (AMB) in human serum and plasma. The procedure involves the addition of the internal standard, p-nitroaniline, to the sample (0.1 ml) followed by protein precipitation with acetonitrile. The supernatant is injected directly onto a C8 chromatographic column and eluted with an acetonitrile-aqueous 0.01 M sodium acetate buffer, pH 7.4, mobile phase. A spectrophotometric detector operated at 405 nm is used. Retention times for internal standard and AMB are 5.2 and 6.6 min, respectively. The assay standard curve is linear between 0.05-2.0 micrograms cm-3. Within- and between-run relative standard deviations (RSD) for high and low concentrations of the drug are less than 5.30%. Analytical recovery of added AMB in serum is 98.4-101.4%. Data obtained by microbiological assay correlated well (r = 0.936) with LC results. Some commonly co-administered drugs and high concentrations of bilirubin are shown not to interfere.
authors.affiliation: Central Laboratory for Clinical Investigation, Osaka University Hospital, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Hosotsubo
authors.forename: K
authors.initials: K
authors.lastname: Hosotsubo
mesh_terms: D000666:Amphotericin B / Q000097:blood*; D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid; D006801:Humans
chemical_list: D000666:Amphotericin B