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Determination of ergot alkaloids in animal feed by liquid chromatography tandem mass spectrometry

Ergot alkaloids are mycotoxins produced by fungi of the Claviceps species, mainly Claviceps purpurea, which can infect different cereals such as rye, wheat, barley and corn. Since these fungi are widespread, there is great risk of food and feed contamination. Ergot alkaloid poisoning causes a disease ergotism, and some of the symptoms are abdominal pain, nausea, hallucinations and gangrene. Among more than 40 known ergot alkaloids, most common ones are ergometrine, ergotamine, ergosine, ergocristine, ergocryptine, ergocornine, and their corresponding epimers. No maximum levels of ergot alkaloids in food and feed have been legislated, although acute reference dose for the sum of the ergot alkaloids is set at 1 µg kg‒1 body weight. There is ongoing research on their presence in food and feed, and enactment of legislation for maximum levels of ergot alkaloids follows in near future. In this work the LC-MS/MS method for determination of 6 most common ergot alkaloids and their epimers in feed is presented. Institute of food safety (RIKILT) has conducted inter-laboratory validation study in order to standardize the method of analysis. Feed samples with known concentrations of ergot alkaloids were dispatched to all participants, including Reference Laboratory for Mycotoxins as part of Andrija Stampar Teaching Institute of Public Health. Samples were extracted with mixture of methanol and water acidified with formic acid and purified by ultrafiltration. Analytes were quantified by multilevel standard addition. Chromatographic separation was achieved on Gemini C18 column (150 x 20 mm, 3 µm) with mobile phases consisting of 1 mM ammonia in acetonitrile and in water. Results have shown good chromatographic separation and recoveries, as well as low levels of quantification. From these results it can be concluded that this method is suitable for determination of ergot alkaloids in feed, but also in other cereal based products, such as food.

alkaloid epimers ; food and feed contamination ; LC-MS/MS ; mycotoxins

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