Development and validation of a rapid multiplex ELISA for pyrrolizidine alkaloids and their N-oxides in honey and feed |
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Authors: | Michalina Oplatowska Christopher T Elliott Anne-Catherine Huet Mark McCarthy Patrick P J Mulder Christoph von Holst Philippe Delahaut Hans P Van Egmond Katrina Campbell |
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Institution: | 1. Institute for Global Food Security, School of Biological Sciences, Queen’s University Belfast, David Keir Building, Stranmillis Road, Belfast, BT9 5AG, UK 2. Département Santé, Centre d’Economie Rurale (CER Groupe), Rue du Point du Jour 8, 6900, Marloie, Belgium 3. RIKILT-Institute of Food Safety, Wageningen UR, P.O. Box 230, 6700 AE, Wageningen, The Netherlands 4. European Commission, DG Joint Research Centre, Institute for Reference Materials and Measurements, 2440, Geel, Belgium
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Abstract: | Pyrrolizidine alkaloids (PAs) are a group of plant secondary metabolites with carcinogenic and hepatotoxic properties. When PA-producing plants contaminate crops, toxins can be transferred through the food chain and cause illness in humans and animals, most notably hepatic veno-occlusive disease. Honey has been identified as a direct risk of human exposure. The European Food Safety Authority has recently identified four groups of PAs that are of particular importance for food and feed: senecionine-type, lycopsamine-type, heliotrine-type and monocrotaline-type. Liquid or gas chromatography methods are currently used to detect PAs but there are no rapid screening assays available commercially. Therefore, the aim of this study was to develop a rapid multiplex ELISA test for the representatives of three groups of alkaloids (senecionine, lycopsamine and heliotrine types) that would be used as a risk-management tool for the screening of these toxic compounds in food and feed. The method was validated for honey and feed matrices and was demonstrated to have a detection capability less than 25 μg/kg for jacobine, lycopsamine, heliotrine and senecionine. The zinc reduction step introduced to the extraction procedure allows for the additional detection of the presence of N-oxides of PAs. This first multiplex immunoassay for PA detection with N-oxide reduction can be used for the simultaneous screening of 21 samples for >12 PA analytes. Honey samples (n?=?146) from various origins were analysed for PA determination. Six samples were determined to contain measurable PAs >25 μg/kg by ELISA which correlated to >10 μg/kg by LC-MS/MS. |
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