Section 41
Chapter 40,381

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

Hosotsubo, H.; Hosotsubo, K.

Journal of Pharmaceutical and Biomedical Analysis 7(8): 975-979


ISSN/ISBN: 0731-7085
PMID: 2490106
DOI: 10.1016/0731-7085(89)80022-6
Accession: 040380182

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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.

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