Ciasca Biancamaria, Pascale Michelangelo, Altieri Valerio Guido, Longobardi Francesco, Suman Michele, Catellani Dante, Lattanzio Veronica M T
Institute of Sciences of Food Production, National Research Council of Italy, Bari, Italy.
Department of Chemistry, University "Aldo Moro" of Bari, Bari, Italy.
J Mass Spectrom. 2018 Sep;53(9):743-752. doi: 10.1002/jms.4089. Epub 2018 Jul 10.
A strong trend toward using highly selective mass spectrometry technologies for screening of multiple mycotoxins has been observed in recent years. In the present study, the process of validation of a multimycotoxin screening method based on liquid chromatography-high-resolution mass spectrometry method is presented. The method was intended for the simultaneous screening of the major Fusarium toxins (deoxynivalenol, 3- and 15-acetyl deoxynivalenol, T-2 and HT-2 toxins, zearalenone, enniatins A, A1, B, and B1, and beauvericin) in wheat. The sample preparation protocol was based on a double extraction (methanol followed by acetonitrile/water mixture) and purification through solid-phase extraction C18 column. To provide insights for full exploitation of the potential of the double-stage high-resolution mass spectrometry detection, a full-scan acquisition event followed by a sequence of 5 fragmentation events (variable data-independent acquisition) was set for mycotoxin detection, the latter to be exploited for confirmatory purposes. Method analytical performances were evaluated through in-house validation and small-scale interlaboratory study, designed according to Commission Regulation 519/2014/EU, setting performance requirements for screening methods for mycotoxins. Screening target concentrations were close to European Union maximum permitted or indicative levels. The in-house validation provided the precision of the response under repeatability conditions and the intermediate precision (both resulting lower than 30%), the cutoff value, and the rate of false suspect results for negative (free of the mycotoxin of interest) samples, which resulted lower than 0.1% in all cases. The collaborative study provided reproducibility and laboratory independent cutoff values. Analysis of reference materials proved method trueness and suitability for screening of the major Fusarium mycotoxins in wheat. Finally, the applicability of the full-scan/variable data-independent acquisition detection approach was successfully tested on a set of naturally contaminated wheat samples, where 2 characteristic product ions could be detected for all identified mycotoxins even at levels in the low μg/kg range.
近年来，人们观察到一种强烈的趋势，即使用高选择性质谱技术来筛查多种霉菌毒素。在本研究中，介绍了一种基于液相色谱 - 高分辨率质谱法的多霉菌毒素筛查方法的验证过程。该方法旨在同时筛查小麦中的主要镰刀菌毒素（脱氧雪腐镰刀菌烯醇、3 - 乙酰脱氧雪腐镰刀菌烯醇和15 - 乙酰脱氧雪腐镰刀菌烯醇、T - 2毒素和HT - 2毒素、玉米赤霉烯酮、恩镰孢菌素A、A1、B和B1以及白僵菌素）。样品制备方案基于双重萃取（先甲醇萃取，然后用乙腈/水混合物萃取），并通过固相萃取C18柱进行净化。为了深入了解充分利用双级高分辨率质谱检测的潜力，设置了一个全扫描采集事件，随后是一系列5次碎裂事件（可变数据非依赖采集）用于霉菌毒素检测，后者用于确证目的。通过内部验证和小规模实验室间研究评估了方法的分析性能，该研究根据欧盟委员会法规519/2014/EU设计，设定了霉菌毒素筛查方法的性能要求。筛查目标浓度接近欧盟最大允许水平或指示性水平。内部验证提供了重复性条件下响应的精密度和中间精密度（两者均低于30%）、截断值以及阴性（不含目标霉菌毒素）样品的假阳性结果率，在所有情况下该结果率均低于0.1%。协作研究提供了重现性和实验室独立截断值。参考物质分析证明了该方法的准确性以及用于筛查小麦中主要镰刀菌霉菌毒素的适用性。最后，全扫描/可变数据非依赖采集检测方法的适用性在一组天然污染的小麦样品上成功进行了测试，在该组样品中，即使在低μg/kg范围内的水平下，所有已鉴定的霉菌毒素均可检测到2个特征产物离子。
