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Abstract
Mycotoxins are produced by fungi as secondary metabolites. They often multi-contaminate food and feed commodities posing a health risk to humans and animals. A fast and easy to apply multiplex screening of these commodities could be useful to detect multi-contamination. For this, we developed a semi-quantitative 6-plex immunoassay using a suspension array of paramagnetic colour-coded microspheres combined with imaging planar array detection for the mycotoxins aflatoxin B1, ochratoxin A, zearalenone, deoxynivalenol, T2-toxin, HT-2 toxin and fumonisin B1. Mycotoxin specific monoclonal antibodies were coupled to different sets of microspheres and mycotoxins conjugated to the fluorescent protein R-phycoerythrin served as reporter molecules. Competition between free mycotoxins in the sample and mixed reporter molecules for antibody binding sites on mixed microspheres created a multiplex direct inhibition immunoassay. The reagents were selected for no or low cross-interactions between the assays and cross-reactions with metabolites and possible masked forms were determined. A within-laboratory validation was carried out using blank and spiked barley samples. Furthermore, the 6-plex was used to screen available barley, and malted barley, reference materials. The validation showed very high inter and intra-day precision for all samples with a maximum relative standard deviation value of 10%. The screening assay allows easy and rapid multiplex detection of the target mycotoxins in barley according to EU legislation. With a cut off factor of 50%, based on the EU maximum levels, we were able to screen at 2 µg kg(-1) for aflatoxin B1, 2.5 µg kg(-1) for ochratoxin A, 625 µg kg(-1) for deoxynivalenol, 50 µg kg(-1) for zearalenone, 1000 µg kg(-1) for fumonisin B1 and 25 µg kg(-1) for T-2 toxin. Thanks to the transportable planar array system, the developed 6-plex has potential for future on-site testing. Future implementation of this method as a pre-screening tool, prior to instrumental analysis, is highly attractive since costly LC-MS/MS analysis of samples below the maximum levels can be avoided.
Original language | English |
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Pages (from-to) | 3968-3976 |
Journal | The Analyst |
Volume | 139 |
Issue number | 16 |
DOIs | |
Publication status | Published - 2014 |
Keywords
- flow-cytometric immunoassay
- surface-plasmon resonance
- suspension array
- liquid-chromatography
- fusarium mycotoxins
- multiplex dipstick
- mass-spectrometry
- cereals
- deoxynivalenol
- ochratoxin
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Dive into the research topics of '6-Plex microsphere immunoassay with imaging planar array detection for mycotoxins in barley'. Together they form a unique fingerprint.Projects
- 1 Finished
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Plato: diagnostic multiplex platform animal feed (KB-15-006-008, KB-06-005-033, KB-06-005-018)
1/01/08 → 31/12/13
Project: EZproject