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Abstract
Tropane alkaloids (TAs) are toxic secondary metabolites produced by plants of, inter alia, the genera Datura (thorn apple) and Atropa (deadly nightshade). The most relevant TAs are (–)-L-hyoscyamine and (–)-L-scopolamine, which act as antagonists of acetylcholine muscarinic receptors and can induce a variety of distinct toxic syndromes in mammals (anti-cholinergic poisoning). The European Union has regulated the presence of seeds of Datura sp. in animal feeds, specifying that the content should not exceed 1000 mg kg–1 (Directive 2002/32/EC). For materials that have not been ground, visual screening methods are often used to comply with these regulations, but these cannot be used for ground materials and compound feeds. Immunological assays, preferably in dipstick format, can be a simple and cost-effective approach to monitor feedstuffs in an HACCP setting in control laboratories. So far no reports have been published on immunoassays that are capable of detecting both hyoscyamine and scopolamine with equal sensitivity and that can be used, preferably in dipstick format, for application as a fast screening tool in feed analysis. This study presents the results obtained for the in-house and inter-laboratory validation of a dipstick immunoassay for the detection of hyoscyamine and scopolamine in animal feed. The target level was set at 800 µg kg–1 for the sum of both alkaloids. By using a representative set of compound feeds during validation and a robust study design, a reliable impression of the relevant characteristics of the assay could be obtained. The dipstick test displayed similar sensitivity towards the two alkaloids and it could be concluded that the test has a very low probability of producing a false-positive result at blank level or a false-negative result at target level. The assay can be used for monitoring of TAs in feedstuffs, but has also potential as a quick screening tool in food- or feed-related poisonings.
Acknowledgements
CONffIDENCE is the acronym of the European Union FP7 project ‘CONtaminants in Food and Feed: Inexpensive DEtectioN for Control of Exposure’. The authors would like to acknowledge the support of Albert Swinkels from MasterLab Nutreco (Boxmeer, the Netherlands) in providing the CONffIDENCE blank feed. The authors want to thank Anne-Catherine Huet (CER Groupe, Marloie, Belgium), Katharina Heinrich (FERA, York, UK), Katrina Campbell (Queen’s University Belfast, Belfast, UK), Veronica Lattanzio (CNR-ISPA, Bari, Italy), Hans van Dalen (Masterlab Nutreco, Putten, the Netherlands), Noan Nivarlet (Unisensor, Liège Wandre, Belgium) and Diana Pereboom-de Fauw (RIKILT, Wageningen, the Netherlands) for their participation in the inter-laboratory validation of the TA dipstick assay.
Funding
The CONffIDENCE project was financially supported by the European Commission [grant agreement number 211326-collaborative project].
Supplemental data
Supplemental data for this article can be accessed here: http://dx.doi.org/10.1080/19440049.2014.914249