Improved high-performance liquid chromatographic determination of Amphotericin B in human serum and plasma

✍ Scribed by Hideo Hosotsubo; Kikumi Hosotsubo

• Publisher: Elsevier Science
• Year: 1989
• Tongue: English
• Weight: 347 KB
• Volume: 7
• Category: Article
• ISSN: 0731-7085
• DOI: 10.1016/0731-7085(89)80022-6

✦ Synopsis

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.