engleski

Mycotoxin analytics – importance of chromatography in UHPLC-MS/MS method development

Geographical distribution and (co)occurrence of mycotoxins, secondary fungal metabolites, in certain food- and feedstuff is greatly influenced by year- to-year varying weather conditions and climate change effects, i.e. extreme changes in temperature and precipitation. Since mycotoxins are known to cause diseases (mycotoxicoses), and thus are regulated by the EU legislation, constant control of their levels is necessary to ensure food and feed safety. Accordingly, there is a need for highly sensitive methods and techniques, such as UHPLC-MS/MS, to unambiguously determine these contaminants. One of the most important steps in development of UHPLC-MS/MS based methods is finding the most suitable chromatographic conditions, i.e. UHPLC column, along with mobile phase combination and proper gradient conditions to provide optimal chromatographic separation and peak shapes, having the effect on method performance characteristics. In this work, three reversed-phase C18 UHPLC columns obtained from different manufacturers were tested using the same mobile phases and elution gradient to choose the best for further optimization. Matrix of extracted cereal sample spiked with all EU regulated mycotoxins was used for the investigation by injection into an UHPLC-MS/MS system, after which peak shape and obtained response were observed for each analyte. Waters Acquity HSS T3 column (1.8 μm, 2.1 x 100 mm) proved to be very useful in the separation of polar, early eluting compounds as it gave the best peak shape for all analytes despite their chemical diversity, using aqueous solution of 5 mM ammonium formate and methanol at a constant flow rate, enabling required limits of detection and quantification.

mycotoxins ; climate change ; food and feed safety ; method development ; chromatography

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