A SPE-UHPLC-DAD method to study the role of fumagillin in the propagation of invasive aspergillosis

Invasive aspergillosis caused by Aspergillus fumigatus is a threat of immunocompromised patients. According to recent studies, fumagillin, a mycotoxin produced by this fungus, has been associated with the propagation of the disease. Therefore, the determination of this molecule might help to understand the mechanisms of this disease and to study the use of fumagillin as a potential biomarker of invasive aspergillosis. In spite of the relevance of fumagillin analysis in microbiological research, no quantitative method has been developed so far for its determination in cell culture media. Here, we present the first validated method for the quantitative analysis of fumagillin in RPMI-1640. The sample treatment consists of a mixed-mode anion exchange Solid Phase Extraction that effectively removes potential EKAIA (202x), artikulua prentsan/article in press. https://doi.org/10.1387/ekaia.23368 Ane Yaldebere eta lankideak Behin-behineko bertsioa (euskara-orrazketaren faltan). 2 interferences and offered a recovery of 83 ± 7%. The analysis was carried out by Ultra High Performance Liquid Chromatography coupled to Diode Array Detection at 336 nm. The method fulfilled all the validation criteria established by EMA (European Medicine Agency) and FDA (Food and Drug Administration) guidelines for bioanalysis. Finally, the method was satisfactorily applied to the quantification of the fumagillin produced by different strains of Aspergillus fumigatus and it was observed that they had a different micotoxin production capacity.