K. (2017). Development of a nanoarray capable of the rapid and simultaneous detection of zearalenone, T2-toxin and fumonisin. Talanta, 164, 368-376. DOI: 10.1016/j.talanta.2016 Published in: Talanta Document Version: Peer reviewed version Queen's University Belfast -Research Portal: Link to publication record in Queen's University Belfast Research Portal Publisher rights © 2017 Elsevier Ltd. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/bync-nd/4.0/ which permits distribution and reproduction for non-commercial purposes, provided the author and source are cited. General rightsCopyright for the publications made accessible via the Queen's University Belfast Research Portal is retained by the author(s) and / or other copyright owners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with these rights.Take down policy The Research Portal is Queen's institutional repository that provides access to Queen's research output. Every effort has been made to ensure that content in the Research Portal does not infringe any person's rights, or applicable UK laws. If you discover content in the Research Portal that you believe breaches copyright or violates any law, please contact openaccess@qub.ac.uk. Fusarium mycotoxins such as trichothecenes, zearalenone and fumonisins occur on a worldwide basis in cereal grains, animal feeds and forages. Practical solutions for multiple mycotoxin determination in samples are required by industry and regulators for cost effective screening purposes. The feasibility of developing a novel multiplex nanoarray for the simultaneous and semi-quantitative detection of three regulated mycotoxins: zearalenone (ZEA), T2-toxin (T2) and fumonisin B1 (FUM) was examined. Additionally, the assay was also able to detect HT2 toxin and fumonisin B2 and B3 due to the cross reactivity profiles of the antibodies used. Individual mycotoxin conjugates specific to the three mycotoxins were nano-spotted onto wells of a microtitre plate. Optimisation of assay parameters and antibodies was undertaken with both individual and multiplex calibration curves generated. A competitive assay format was employed enabling a calibration curve for concentration analysis and duplicate results for up to 40 samples in 70 min for the three target mycotoxins. The characteristics and performance of the nanoarray were evaluated including sensitivity and specificity for each target. Additionally, intra and inter spotting precision, cross reactivity, matrix effects and sample analysis in maize and wheat (n=8) was performed. Sensitivity, determined as the concentration causing 50 % inhibition, was 70.1, 2.8 and 90.9 ppb in PBS, 172.4, 3.2 and 129.3 ppb in methanol, 197.4, 0.7 and 216.7 ppb in wheat and 43.6, 0.5 and 25.9 ppb in maize for ZEA, T2 and FUM respectively. Intra spotting precision was 6, 11 and 10 % for PBS and 5, 11 and 12 % for methanol for ZEA, T2 and FUM respectively. Inter spotti...

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Background Aflatoxin M1 (AFM1) is the major metabolite of Aflatoxin B1 (AFB1) and can be found in the milk of animals fed with feed containing AFB1. The frequency of occurrence of AFM1 in milk has led to the development of specific quantitative methods of analysis to mitigate the risk of adversely affect human health. Objective: The objective was to demonstrate that I’screen AFLA M1 Milk ELISA kit can quantify AFM1 in raw bovine milk and in powdered milk. Methods Assay performance was evaluated studying lot-to-lot consistency, assay stability, robustness and possible interferences of related molecules. Raw bovine milk samples spiked at 0, 5.0, 20, 50, 100, 200 ng/L of AFM1 and powdered milk reference materials and spiked samples at 100 and 200 ng/L were tested to determine recovery, repeatability and bias. LOD and LOQ were also determined for both matrixes. Results: High selectivity for AFM1 was demonstrated and performances were consistent, robust and stable. The LOQ was validated at 5 ng/L for raw milk and 50 ng/L for powdered milk. Recoveries for spiked raw and powdered milk were 97–122%, with RSDr < 10%, and 106–111% for reference materials, with RSDr < 5%. Conclusions: The data collected validate the method as a selective, specific, sensitive, accurate and precise tool for the analysis of AFM1 in raw bovine milk and powdered milk. Highlights: We demonstrated that I’screen AFLA M1 is a reliable kit and a proper screening tool suitable for high analytical throughputs.