HPLC assay for determination of amphotericin B in biological samples.
A fast and selective HPLC method for assaying amphotericin B in biological samples was developed and validated. The chromatographic separation was achieved in less than 12 min on a reverse-phase C(18) column using an acetonitrile-acetic acid-water (52:4.3:43.7, v/v/v) mixture as mobile phase. The flow rate was 1 mL/min and the effluent was monitored at 406 nm. A linear response over the concentration range 0.1-10.0 microg/mL was obtained. Intra-day and inter-day RSDs were below 5% for all the sample types. This new HPLC method was applied to assay amphotericin B in plasma and several tissue samples such as kidney, liver, spleen and bone marrow. Application of this method to pharmacokinetic studies in mice and dog is provided.
Espada
R
R
Department of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, Complutense University, 28040 Madrid, Spain.
Josa
J M
JM
Valdespina
S
S
Dea
M A
MA
Ballesteros
M P
MP
Alunda
J M
JM
Torrado
J J
JJ
D000666:Amphotericin B / Q000032:analysis* / Q000097:blood* / Q000493:pharmacokinetics; D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004285:Dogs; D005260:Female; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D051379:Mice; D015203:Reproducibility of Results; D013154:Spleen / Q000378:metabolism
D000666:Amphotericin B