Development and validation of a rapid multiplex ELISA for pyrrolizidine alkaloids and their N-oxides in honey and feed.

Oplatowska, Michalina; Elliott, Christopher; Huet, Anne-Catherine; McCarthy, Mark; Mulder, Patrick; Holst, Christoph; Delahaut, Philippe; Egmond, Hans; Campbell, Katrina

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.

ENZYME-linked immunosorbent assay; PYRROLIZIDINES; PLANT metabolites; HEPATOTOXICOLOGY; CARCINOGENICITY testing; LIQUID chromatography; GAS chromatography; EUROPEAN Food Safety Authority

Analytical & Bioanalytical Chemistry, 2014, Vol 406, Issue 3, p757

1618-2642

Academic Journal

10.1007/s00216-013-7488-7