Amphotericin B determination in respiratory secretions by reversed-phase liquid chromatography.
Direct delivery of amphotericin B (AMB) to the respiratory tract may be an alternative to intravenous administration. The use of inhalation allows high AMB concentrations to be achieved at the site of infection. A reversed-phase high-performance liquid chromatographic method with a 30-mm-long column is described for assaying AMB in respiratory secretions obtained by bronchoaspiration (BAS) and bronchoalveolar lavage (BAL). Sample clean-up involved treatment with methanol (BAS) and solid-phase extraction onto Sep-Pak C18 cartridges (BAL). The mobile phase consisted of 2.5 mM Na2EDTA-acetonitrile (70:30, v/v). The retention time of AMB was 1.5 min. The range of the assay was from 0.1 to 5 micrograms/ml. The mean recovery was over 90% for both fluids. Within-day and between-day RSDs ranged from 3.10 to 11.87%. AMB in the BAS samples was stable for two days at 20-25 degrees C and for three months at -20 degrees C. The drug in the BAL fluid was stable for one day at 20-25 degrees C, seven days at 4 degrees C and for one month at -20 degrees C.
Biochemistry Service, Hospital General Universitario Vall D'Hebrón, Barcelona, Spain.
R
R
Lopez
L
L
Pou
I
I
Andres
V
V
Monforte
A
A
Roman
C
C
Pascual
D000666:Amphotericin B / Q000032:analysis*; D000935:Antifungal Agents / Q000032:analysis*; D001826:Body Fluids / Q000737:chemistry*; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D002853:Chromatography, Liquid; D006801:Humans; D007202:Indicators and Reagents
D000935:Antifungal Agents; D007202:Indicators and Reagents; D000666:Amphotericin B