Soares Ruben R G, Azevedo Ana M, Fernandes Pedro, Chu Virginia, Conde João P, Aires-Barros M Raquel
IBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal; Instituto de Engenharia de Sistemas e Computadores, Microsistemas e Nanotecnologias (INESC MN) and IN, Institute of Nanoscience and Nanotechnology, Lisbon, Portugal.
IBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal; Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
J Chromatogr A. 2017 Aug 18;1511:15-24. doi: 10.1016/j.chroma.2017.07.004. Epub 2017 Jul 5.
The rapid detection of mycotoxins in feed samples is becoming an increasingly relevant challenge for the food production sector, in order to effectively enforce current regulations and assure food and feed safety. To achieve rapid mycotoxin detection, several biosensing strategies have been published, many reaching assay times of the order of a few minutes. However, the vast majority of these rely on sample preparation based on volatile organic solvents, often comprising complex multi-step procedures and devoid of clean-up and/or concentration effects. Here, a novel sample preparation methodology based on a green, non-toxic and inexpensive polyethylene glycol-sodium citrate aqueous two-phase system is reported, providing single-step extraction and concentration of three target mycotoxins within 20min: aflatoxin B1 (AFB1), ochratoxin A (OTA) and deoxynivalenol (DON). With point-of-need applications in mind, the extraction procedure was optimized and validated using a rapid multi-toxin microfluidic competitive immunoassay. The assay was successfully tested with spiked complex solid matrices including corn, soy, chickpea and sunflower-based feeds and limits of detection of 4.6ngg±15.8%, 24.1ngg±8.1% and 129.7ngg±53.1% (±CV) were obtained in corn for AFB1, OTA and DON, respectively. These sensitivities are fit-for-purpose at the required regulatory and recommended limits for animal feed, providing an effective and safe semi-quantitative mycotoxin analysis that can be performed in the field.
为有效执行现行法规并确保食品和饲料安全，饲料样本中霉菌毒素的快速检测正成为食品生产行业日益重要的挑战。为实现霉菌毒素的快速检测，已发表了多种生物传感策略，其中许多检测时间可达几分钟左右。然而，这些策略绝大多数依赖基于挥发性有机溶剂的样品制备，通常包括复杂的多步程序，且缺乏净化和/或浓缩效果。在此，报道了一种基于绿色、无毒且廉价的聚乙二醇-柠檬酸钠水两相系统的新型样品制备方法，可在20分钟内对三种目标霉菌毒素进行单步提取和浓缩：黄曲霉毒素B1（AFB1）、赭曲霉毒素A（OTA）和脱氧雪腐镰刀菌烯醇（DON）。考虑到即时检测应用，使用快速多毒素微流控竞争免疫分析法对提取程序进行了优化和验证。该分析方法在添加了毒素的复杂固体基质（包括玉米、大豆、鹰嘴豆和向日葵基饲料）上成功进行了测试，在玉米中分别获得了AFB1、OTA和DON的检测限为4.6ng/g±15.8%、24.1ng/g±8.1%和129.7ng/g±53.1%（±CV）。这些灵敏度符合动物饲料所需的监管和推荐限量要求，可在现场进行有效且安全的半定量霉菌毒素分析。
