In this study, 120 silage samples collected in 2015 from farms in Poland were analysed by a multimycotoxin method based on liquid chromatography coupled with tandem mass spectrometry. The study included toxins which are regulated within the European Union (aflatoxins, deoxynivalenol, fumonisins, T-2/HT-2 toxins, ochratoxin A and zearalenone) and non-regulated mycotoxins (enniatins, beauvericin, 8-ketotrichothecenes, sterigmatocystin, zearalenone derivatives). All silage samples were positive for at least one mycotoxin, and 61% of samples contained five or more mycotoxins simultaneously. The most frequently detected toxins were deoxynivalenol, nivalenol, zearalenone, enniatins and beauvericin, although the levels of these toxins were relatively low. The mean concentration of deoxynivalenol and zearalenon was 406 and 80.6 μg/kg, respectively, and two toxins were positive-correlated. This is the first study that provides information about emerging mycotoxins contaminating silage in Poland.

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Trichothecene toxins are confirmed or suspected virulence factors of various plant-pathogenic Fusarium species. Plants can detoxify these to a variable extent by glucosylation, a reaction catalyzed by UDP-glucosyltransferases (UGTs). Due to the unavailability of analytical standards for many trichothecene-glucoconjugates, information on such compounds is limited. Here, the previously identified deoxynivalenol-conjugating UGTs HvUGT13248 (barley), OsUGT79 (rice) and Bradi5g03300 (Brachypodium), were expressed in E. coli, affinity purified, and characterized towards their abilities to glucosylate the most relevant type A and B trichothecenes. HvUGT13248, which prefers nivalenol over deoxynivalenol, is also able to conjugate C-4 acetylated trichothecenes (e.g., T-2 toxin) to some degree while OsUGT79 and Bradi5g03300 are completely inactive with C-4 acetylated derivatives. The type A trichothecenes HT-2 toxin and T-2 triol are the kinetically preferred substrates in the case of HvUGT13248 and Bradi5g03300. We glucosylated several trichothecenes with OsUGT79 (HT-2 toxin, T-2 triol) and HvUGT13248 (T-2 toxin, neosolaniol, 4,15-diacetoxyscirpenol, fusarenon X) in the preparative scale. NMR analysis of the purified glucosides showed that exclusively β-d-glucosides were formed regio-selectively at position C-3-OH of the trichothecenes. These synthesized standards can be used to investigate the occurrence and toxicological properties of these modified mycotoxins.

QuEChERS and zirconium-coated silica SPE in a multiresidue method for the analysis of veterinary drug residues and other contaminates in eggs by LC-MS/MS.