Dairy Sciences, Institute of Veterinary Food Science, Justus-Liebig-University Giessen, Ludwigstrasse 21, 35390, Giessen, Germany.
Department Safety and Quality of Milk and Fish Products, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, Hermann-Weigmann-Strasse 1, 24103, Kiel, Germany.
Mycotoxin Res. 2022 Nov;38(4):265-274. doi: 10.1007/s12550-022-00467-x. Epub 2022 Sep 2.
Plant-based milk alternatives (PBMAs) are a potential source of mycotoxin uptake. To ensure food safety, simple and rapid testing methods of PBMAs for mycotoxins are therefore required. This study investigated the applicability of enzyme immunoassay (EIA) methods for direct testing of PBMAs without sample extraction. Mycotoxin analyses included aflatoxin B (AFB), sterigmatocystin (STC), ochratoxin A (OTA), deoxynivalenol (DON), and T-2/HT-2-toxin (T-2/HT-2). It was found that the PBMA matrix negatively affected the EIA to varying degrees, thus affecting the reliability of the results. A dilution of PBMAs of at least 1:8 was necessary to overcome matrix interference. This resulted in calculated detection limits of 0.4 µg/L (AFB), 2 µg/L (STC), 0.08 µg/L (OTA), 16 µg/L (DON), and 0.4 µg/L (T-2/HT-2). After analysis of 54 PBMA products from German retail stores, positive results in at least one test system were obtained for 23 samples. However, most positive results were near the calculated detection limit. Control analyses of selected samples by LC-MS/MS for AFB, STC, and OTA qualitatively confirmed the presence of trace amounts of STC in some samples, but quantitative agreement was poor. It was concluded that the high diversity of ingredients used in PBMAs led to a highly variable degree of sample matrix interference even in a 1:8 dilution. Since the use of higher dilutions conflicts with the need to achieve low detection limits, the application of EIA for routine mycotoxin analysis in PBMA for mycotoxins requires further study on the development of a feasible sample preparation method.
植物基奶替代品（PBMAs）是真菌毒素摄入的潜在来源。因此，为了确保食品安全，需要开发简单、快速的 PBMAs 真菌毒素检测方法。本研究探讨了酶联免疫吸附测定（EIA）方法在无需样品提取的情况下直接检测 PBMAs 的适用性。真菌毒素分析包括黄曲霉毒素 B（AFB）、桔青霉素（STC）、赭曲霉毒素 A（OTA）、脱氧雪腐镰刀菌烯醇（DON）和 T-2/HT-2 毒素（T-2/HT-2）。结果表明，PBMAs 基质对 EIA 产生不同程度的负面影响，从而影响结果的可靠性。需要将 PBMAs 稀释至少 1:8 以克服基质干扰。这导致计算出的检测限分别为 0.4 µg/L（AFB）、2 µg/L（STC）、0.08 µg/L（OTA）、16 µg/L（DON）和 0.4 µg/L（T-2/HT-2）。对德国零售店的 54 种 PBMAs 产品进行分析后，至少有 23 种样品在至少一个测试系统中得到阳性结果。然而，大多数阳性结果接近计算出的检测限。对部分样品采用 LC-MS/MS 进行 AFB、STC 和 OTA 的控制分析定性证实了一些样品中存在痕量 STC，但定量结果一致性较差。结果表明，PBMAs 中使用的成分种类繁多，即使在 1:8 稀释的情况下，样品基质干扰的程度也存在很大差异。由于使用更高的稀释度与达到低检测限的需求相冲突，因此，EIA 应用于 PBMAs 中真菌毒素的常规真菌毒素分析需要进一步研究可行的样品制备方法的开发。
