Fumagilina mikotoxinak aspergilosi inbaditzailearen garapenean duen rola aztertzeko SPE-UHPLC-DAD metodo analitikoa: A SPE-UHPLC-DAD method to study the role of fumagillin in the propagation of invasive aspergillosis

Abstract

Invasive aspergillosis caused by Aspergillus fumigatus is a threat of immunocompromised patients. Lately, 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 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 and FDA guidelines for bioanalysis and was satisfactorily applied to the quantification of the fumagillin produced by different strains of Aspergillus fumigatus with different toxin production capacity.