Employing immuno-affinity for the analysis of various microbial metabolites of the mycotoxin deoxynivalenol


Zhu, Y., Hassan, Y.I., Shao, S., Zhou, T. (2018). Employing immuno-affinity for the analysis of various microbial metabolites of the mycotoxin deoxynivalenol. Journal of Chromatography A, [online] 1556 81-87. http://dx.doi.org/10.1016/j.chroma.2018.04.067

Plain language summary

Deoxynivalenol (DON) or vomitoxin is a widely detected fungal toxin that causes multifaceted problems connected with the accidental contamination of human food and animal feed. Effective and affordable analysis methods of DON in food/feed materials are urgently sought as part of larger controlling strategies. This research paper provides a fast, accurate, and easy-to-operate protocol for capturing this toxin using conjugated antibodies that were commercially produced to target DON followed by the analysis with high performance liquid chromatography. Moreover, the same protocol (and antibody) is shown to be capable of binding other bacterial metabolites of DON some of which are less toxic. The presented results also narrow down the region of physical DON/antibody interactions, which should aid in designing more specific antibodies in the future for the analytical purposes. Furthermore this work highlights the need to consider the binding of bacterial metabolites of DON to the incorporated DON-antibodies (used in commercially available kits) when interrupting the outcomes to avoid any DON overestimations. In essence, this work collectively provides an optimized analytical tool that significantly simplifies the determination of DON (and its bacterial metabolites) in many food/feed matrixes and provides the springboard to further DON detoxification research in the near future.


Deoxynivalenol (DON) is a type B trichothecene mycotoxin that is commonly detected in grains infested with Fusarium species. The maximum tolerated levels of DON in the majority of world's countries are restricted to 0.75 mg kg−1 within the human food chain and to less than 1–5 mg kg−1 in animal feed depending on the feed material and/or animal species due to DON's short and long-term adverse effects on human health and animal productivity. The ability to accurately analyze DON and some of its fungal/bacterial metabolites is increasingly gaining a paramount importance in food/feed analysis and research. In this study, we used the immuno-affinity approach to enrich and detect DON and three of its bacterial metabolites, namely 3-epi-DON, 3-keto-DON, and deepoxy-DON (DOM-1). The optimized enrichment step coupled with high performance liquid chromatography can accurately and reproducibly quantify the aforementioned metabolites in feed matrixes (silage extract as an example in this case). It minimizes any background interface and provides a fast and easy-to-operate protocol for the analytical determination of such metabolites. More importantly, the presented data demonstrates the ability of the utilized monoclonal antibody, generated originally to capture DON in Enzyme-Linked Immunosorbent Assays (ELISA), to cross react with three less/non-toxic DON metabolites. This raises the concerns about the genuine need to account for such cross-reactivity when DON contamination is assessed through an immuno-affinity based analyses using the investigated antibody.

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