title: Development and validation of a rapid multiplex ELISA for pyrrolizidine alkaloids and their N-oxides in honey and feed.
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.
authors: Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, David Keir Building, Stranmillis Road, Belfast, BT9 5AG, UK.
authors: Michalina
authors: M
authors: Oplatowska
authors: Christopher T
authors: CT
authors: Elliott
authors: Anne-Catherine
authors: AC
authors: Huet
authors: Mark
authors: M
authors: McCarthy
authors: Patrick P J
authors: PP
authors: Mulder
authors: Christoph
authors: C
authors: von Holst
authors: Philippe
authors: P
authors: Delahaut
authors: Hans P
authors: HP
authors: Van Egmond
authors: Katrina
authors: K
authors: Campbell
mesh_terms: D000821:Animal Feed / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D006722:Honey / Q000032:analysis*; D057230:Limit of Detection; D010087:Oxides / Q000032:analysis* / Q000737:chemistry; D011763:Pyrrolizidine Alkaloids / Q000032:analysis*; D013997:Time Factors
chemical_list: D010087:Oxides; D011763:Pyrrolizidine Alkaloids