Institute of Bioanalytics and Agro-Metabolomics, Department of Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences, Vienna (BOKU), Konrad Lorenz Str. 20, 3430 Tulln, Austria.
Institute of Microbial Genetics, Department of Applied Genetics and Cell Biology, BOKU, Konrad Lorenz Str. 24, 3430 Tulln, Austria.
Toxins (Basel). 2021 Aug 27;13(9):600. doi: 10.3390/toxins13090600.
The Fusarium mycotoxin deoxynivalenol (DON) is a common contaminant of cereals and is often co-occurring with its modified forms DON-3-glucoside (D3G), 3-acetyl-DON (3ADON) or 15-acetyl-DON (15ADON). A stable-isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) based method for their determination in cereals was developed and validated for maize. Therefore, C-labelled D3G was enzymatically produced using C-DON and [CGlc]-sucrose and used as an internal standard (IS) for D3G, while uniformly C labelled IS was used for the other mycotoxins. Baseline separation was achieved for the critical peak pair DON/D3G, while 3ADON/15ADON could not be fully baseline separated after testing various reversed phase, fluorinated phase and chiral LC columns. After grinding, weighing and extracting the cereal samples, the raw extract was centrifuged and a mixture of the four C-labelled ISs was added directly in a microinsert vial. The subsequent analytical run took 7 min, followed by negative electrospray ionization and selected reaction monitoring on a triple quadrupole MS. Maize was used as a complex cereal model matrix for validation. The use of the IS corrected the occurring matrix effects efficiently from 76 to 98% for D3G, from 86 to 103% for DON, from 68 to 100% for 15ADON and from 63 to 96% for 3ADON.
镰刀菌真菌毒素脱氧雪腐镰刀菌烯醇（DON）是谷物的常见污染物，通常与它的修饰形式 DON-3-葡萄糖苷（D3G）、3-乙酰-DON（3ADON）或 15-乙酰-DON（15ADON）共同存在。建立并验证了用于谷物中这些化合物测定的基于稳定同位素稀释液相色谱-串联质谱（LC-MS/MS）的方法，该方法针对玉米。因此，使用 C-DON 和 [CGlc]-蔗糖通过酶法生产了 C 标记的 D3G，并将其用作 D3G 的内标（IS），而其他真菌毒素则使用均一 C 标记的 IS。在测试了各种反相、氟化相和手性 LC 柱后，实现了 DON/D3G 关键峰对的基线分离，而 3ADON/15ADON 则无法完全基线分离。谷物样品经过研磨、称重和提取后，将原始提取物离心，并直接在微型插入小瓶中加入四种 C 标记 IS 的混合物。随后的分析运行时间为 7 分钟，然后在三重四极杆 MS 上进行负离子电喷雾电离和选择反应监测。玉米被用作复杂谷物模型基质进行验证。使用 IS 可有效地校正发生的基质效应，使 D3G 的校正回收率从 76%提高到 98%，使 DON 的校正回收率从 86%提高到 103%，使 15ADON 的校正回收率从 68%提高到 100%，使 3ADON 的校正回收率从 63%提高到 96%。
