Development, validation and application of an LC-MS/MS based method for the determination of deoxynivalenol and its conjugates in different types of beer

Abstract

After water and tea, beer is the third most popular beverage worldwide. Brewed from malted cereal grains, beer is known to be potentially contaminated with mycotoxins. Some studies have shown that not only the Fusarium mycotoxins deoxynivalenol (DON) and 3-acetyl-DON (3-ADON), but also the conjugated mycotoxin deoxynivalenol-3-glucoside (D3G) can be found in beer on a regular basis, albeit usually at low concentrations. The aim of this work was to develop the first triple quadrupole LC-MS/MS based method for the determination of DON, D3G and 3-ADON in beer and to perform an in-house validation. The simple sample preparation includes degassing, precipitation of matrix compounds and reconstitution of the dried-down sample in solvent. Since different kinds of beer exist and method performance parameters will likely differ, we categorised the samples into pale, wheat, dark, bock and non-alcoholic beers, as well as shandies, and validated all six matrices. Although three individual beers for each category were spiked at eight levels prior to sample preparation, the repeatability of the overall method was still excellent with relative standard deviations from 4-16% for all analytes and types of beer. Limits of detection were in the sub- or low-μg/kg range. Apparent recoveries of 60-90% for DON, 39-69% for D3G and 96-124% for 3-ADON were obtained for the different types of beer, with dark and bock beers being the most difficult matrices. To prove the applicability of the method, ten beers of each category were analysed. While average concentrations of 6.6 μg/l for DON and D3G were found, no 3-ADON was detected in any of the samples.