Evidence that impurities contribute to the fluorescence of the polyene antibiotic amphotericin B

Objectives: Based on the assertion that fluorescence spectroscopy detects dimers of the polyene antibiotic amphotericin B (AmB), this technique was recently proposed to analyse the interaction of the drug with cell membranes. However, contradictory results indicate that this ‘dimeric’ fluorescence might actually originate from polyene impurities. We used a highly purified AmB to challenge this last proposal. Methods: Comparison of the fluorescence of AmB from different origins was made in dimethyl sulphoxide (DMSO); concentration and sodium dodecyl sulphate (SDS) addition dependencies were analysed in water. Results: Excitation of fluorescence in the absorption band of the AmB monomer (around 410 nm) revealed no difference between the different samples, in contrast with what was observed by excitation in the absorption wavelengths of self-associated AmB (around 325 nm). Furthermore, in this latter case, no concentration dependence was observed, in DMSO or in water. SDS addition increased the fluorescence in water. Conclusions: The fluorescence of AmB observed by excitation in the absorption wavelengths of selfassociated species (around 325 nm) is explainable by the presence of impurities. Fluorescence is probably not appropriate for characterization of the drug interaction with cell membranes.