Ultra-performance liquid chromatography for quantification of amphotericin B plasma concentrations after use of liposomal amphotericin B

Ruth Van Daele*, Yvo de Beer, Sander Croes, Rob Aarnoutse, Joost Wauters, Johan Maertens, Isabel Spriet, Roger J. Bruggemann

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Web of Science)


Objectives: Liposomal amphotericin B is widely used to treat Life-threatening invasive fungal infections and has replaced conventional amphotericin B deoxycholate due to its more favourable toxicity profile. Despite the fact that Liposomal amphotericin B has been Licensed for several decades, there is still a paucity of clinical pharmacokinetic data. An assay for the quantification of amphotericin B is necessary to allow the study of its pharmacokinetics.

Methods: A UPLC-photodiode array (PDA) analytical method was developed and validated (Linearity, accuracy, precision, dilution integrity, carry-over, selectivity and stability) in accordance with EMA requirements.

Results: The analytical method was validated over a concentration range of 0.5-50.0 mg/L. Accuracy ranged from 97.6% to 112.1% and within-day repeatability and between-day reproducibility from 1.0% to 6.6% and from 0.4% to 4.6%, respectively, dependent on the concentration. Originally, the goal was to develop an analytical method to separate the Liposomal and free amphotericin B fractions, but this was not achieved. Difficulties and bottlenecks encountered are presented.

Conclusions: A UPLC-PDA analytical method was developed to quantify total amphotericin B in plasma after the use of Liposomal amphotericin B.

Original languageEnglish
Pages (from-to)961-966
Number of pages6
JournalJournal of Antimicrobial Chemotherapy
Issue number4
Publication statusPublished - Apr 2021



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