Kovač Marija, Nevistić Ante, Kovač Tihomir, Babić Jurislav, Šarić Antonija, Miličević Borislav, Panjičko Mario, Šarkanj Bojan
Department of Food Technology, University North, Trg Dr. Žarka Dolinara 1, 48000 Koprivnica, Croatia.
Inspecto Ltd., Industrijska Zona Nemetin, Vukovarska Cesta 239b, 31000 Osijek, Croatia.
J Fungi (Basel). 2022 Jun 24;8(7):665. doi: 10.3390/jof8070665.
The availability of reliable sensitive multi-analyte methods for unambiguous determination of mycotoxins is crucial for ensuring food and feed safety, considering their adverse health effects and (co-)occurrence in various foods. Accordingly, a multi-mycotoxin confirmatory method for simultaneous determination of 11 mycotoxins regulated in cereals within the European Union (EU) using ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was developed and in-house validated to fit the EU legislation requirements for analytical methods. A simple sample preparation was based on a solid−liquid extraction using a solvent mixture acetonitrile/water/formic acid (79/20/1, v/v/v) and a dilution of raw extract using water/acetonitrile/formic acid (79/20/1, v/v/v) before instrumental analysis. Average recoveries in all three validated cereal crop types (maize, wheat, and barley), spiked at multiple levels, were found acceptable for all analytes when matrix-matched calibration was used, ranging from 63.2% to 111.2% and also showing very good repeatability, with relative standard deviations below 20%. Matrix effect (SSE) evaluation revealed maize as the most complex of the three analyzed cereal matrices, with strong SSE (<50% and >150%) recorded for all 11 analyzed mycotoxins. An additional method verification was performed through successful participation in proficiency testing schemes, with the achieved z-scores generally in the acceptable range of −2 ≤ z ≤ 2. The obtained validation results demonstrated the suitability of the developed confirmatory multi-mycotoxin UHPLC-MS/MS method based on a dilute-and-shoot principle for the simultaneous determination of low concentrations of 11 EU-regulated mycotoxins in cereals, including aflatoxins B1, B2, G1 and G2, deoxynivalenol, fumonisins B1 and B2, zearalenone, T-2 and HT-2 toxins, and ochratoxin A.
考虑到霉菌毒素对健康的不利影响以及它们在各种食品中的（共同）存在，拥有可靠、灵敏的多分析物方法来明确测定霉菌毒素对于确保食品和饲料安全至关重要。因此，开发了一种使用超高效液相色谱-串联质谱（UHPLC-MS/MS）同时测定欧盟规定的谷物中11种霉菌毒素的多霉菌毒素确证方法，并在内部进行了验证，以符合欧盟分析方法的立法要求。简单的样品制备基于使用乙腈/水/甲酸（79/20/1，v/v/v）的溶剂混合物进行固液萃取，并在仪器分析前用水/乙腈/甲酸（79/20/1，v/v/v）稀释粗提物。当使用基质匹配校准时，在所有三种经过验证的谷物作物类型（玉米、小麦和大麦）中，多个添加水平下的所有分析物的平均回收率均可接受，范围为63.2%至111.2%，并且还显示出非常好的重复性，相对标准偏差低于20%。基质效应（SSE）评估表明，玉米是三种分析谷物基质中最复杂的，所有11种分析的霉菌毒素均记录到强烈的SSE（<50%和>150%）。通过成功参与能力验证计划进行了额外的方法验证，获得的z分数通常在可接受的-2≤z≤2范围内。获得的验证结果表明，基于稀释进样原理开发的确证性多霉菌毒素UHPLC-MS/MS方法适用于同时测定谷物中低浓度的11种欧盟规定的霉菌毒素，包括黄曲霉毒素B1、B2、G1和G2、脱氧雪腐镰刀菌烯醇、伏马毒素B1和B2、玉米赤霉烯酮、T-2和HT-2毒素以及赭曲霉毒素A。
