title: New high-performance liquid chromatographic method for amphotericin B analysis using an internal standard.
abstract: A simple and reproducible HPLC method for the analysis of amphotericin B (AmB) in serum, lung and liver using natamycin as the internal standard was developed. AmB and natamycin were extracted from serum, lung and liver and were separated using an isocratic elution from C18 reversed-phase column. The mobile phase consisted of acetonitrile-10 mM acetate buffer pH 4.0 (37:63, v/v). The HPLC system had two detectors in series. One was set at 303 nm and the other at 383 nm for the detection of natamycin and AmB, respectively. The retention times of AmB and natamycin were 15 and 6 min, respectively. The recovery efficiency was 96%-70%. The limit of quantification was 0.1 microgram/ml. The assay was reproducible, the within-day coefficient of variation (n = 6) was < 8% for serum, lungs and liver. The between-day variability (n = 6) was < 7.7% for serum, liver and lungs at 1 microgram/ml or 1 microgram/g tissue concentration. The assay was linear within the range 1-40 micrograms/ml (r2 = 0.99).
authors.affiliation: University of Oklahoma Health Science Center, College of Pharmacy, Oklahoma City 73190, USA.
authors.forename: M P
authors.initials: MP
authors.lastname: Lambros
authors.forename: S A
authors.initials: SA
authors.lastname: Abbas
authors.forename: D W
authors.initials: DW
authors.lastname: Bourne
mesh_terms: D000666:Amphotericin B / Q000032:analysis* / Q000097:blood; D000818:Animals; D000935:Antifungal Agents / Q000032:analysis* / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008099:Liver / Q000737:chemistry; D008168:Lung / Q000737:chemistry; D051379:Mice; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: D000935:Antifungal Agents; D000666:Amphotericin B