LC-MS Based Method of Analysis for the Simultaneous Determination of four Mycotoxins in Cereals and Feed: Results of a Collaborative
Study
An LC-MS/MS based method of analysis to determine the four Fusarium toxins deoxynivalenol, HT-2 toxin, T-2 toxin, and zearalenone in cereals and cereal-based compound animal feed has been validated through a collaborative study. After extraction of the mycotoxins with ethyl acetate / water, and addition of sodium sulphate an aliquot of the organic phase was spiked with stable-isotope labelled isotopologues of the targeted analytes and dried down. The dry extract was then reconstituted with mobile phase and injected into a LC-MS. The described use of the isotopologues keeps costs down while still offering many of their benefits. This is evidenced by relative repeatability standard deviations (RSDr) between 5 and 15 %. Exceptions were T-2 toxin at 7 μg/kg with 27%, and at 3.5 μg/kg with 35%, and zearalenone at 3.4 μg/kg with 32% RSDr.
The tested contamination ranges were 88 to 559 μg/kg for deoxynivalenol, 22 to 178 μg/kg for HT-2 toxin, 3.5 to 50 μg/kg for T-2 toxin, and 3.4 to 430 μg/kg for zearalenone. For 10 of the 20 analyte / matrix combinations (four analytes in five matrices) Horwitz ratios between 0.6 and 0.9 were computed, for another six the ratios were below 1.5. The remaining four test samples were associated with Horwitz ratios between 2.0 and 4.4. They were the samples described above, two containing T-2 toxin and one zearalenone, plus one complex matrix sample containing zearalenone at a low contamination level. For this complex matrix sample we were able to show the importance of proper separation in LC-MS.
Because of the use of test materials having assigned reference values in this study trueness could be assessed. The observed biases were small and only significant for deoxynivalenol (-8%) and HT-2 toxin (-11%). For T-2 toxin and zearalenone they were insignificant. To facilitate the checking of compliance of a test result produced with this method with legislation a description on how to estimate measurement uncertainty based on these results is provided.
All of the above shows that the studied method is fit for the purpose of enforcing existing and anticipated legislative limits of the four Fusarium toxins deoxynivalenol, HT-2 toxin, T-2 toxin, and zearalenone in unprocessed cereals and cereal-based compound animal feed.
BREIDBACH Andreas;
BOUTEN Katrien;
KROEGER-NEGOITA Katy;
STROKA Joerg;
ULBERTH Franz;
2013-03-22
Publications Office of the European Union
JRC80176
978-92-79-28937-8,
1831-9424,
EUR 25853,
OP LA-NA-25853-EN-N,
https://publications.jrc.ec.europa.eu/repository/handle/JRC80176,
10.2787/77845,
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