Department of Pharmacology and Toxicology, National Veterinary Research Institute, Al. Partyzantów 57, 24-100 Puławy, Poland.
Toxins (Basel). 2020 Jun 1;12(6):362. doi: 10.3390/toxins12060362.
A liquid chromatography-tandem mass spectrometry method was developed for simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3Ac-DON), 15-acetyldeoxynivalenol (15Ac-DON), DON-3-glucoside (DON-3Glc) nivalenol and fusarenone-X in feedstuffs. Different techniques of sample preparation were tested: solid-liquid-extraction, QuEChERS, solid phase extraction with OASIS HLB columns or immunoaffinity columns and a Mycosep 225 Trich column. None of the six immunoaffinity columns tested showed cross-reactivity to all of the mycotoxins. Surprisingly, the results show that if the immunoaffinity columns bound 3Ac-DON, then they did not bind 15Ac-DON. The most efficient sample preparation was achieved with a Mycosep 225 Trich column clean-up. The chromatography was optimised to obtain full separation of all analytes (including 3Ac-DON and 15Ac-DON isomeric form). The validation results show the relative standard deviations for repeatability and reproducibility varied from 4% to 24%. The apparent recovery ranged between 92% and 97%, and the limit of quantification described a 1.30 to 50 µg/kg range. The method trueness was satisfactory, as assessed by a proficiency test and analysis of reference material. A total of 99 feed samples were analysed by the developed method, revealing the presence of DON and DON-3Glc in 85% and 86% of examined animal feeds, respectively at concentrations between 1.70 and 1709 µg/kg. The ratios DON-3Glc to DON in the surveyed feedstuffs were from a low of 3% to high of 59%.
建立了一种液相色谱-串联质谱法，用于同时测定饲料中的脱氧雪腐镰刀菌烯醇（DON）、3-乙酰脱氧雪腐镰刀菌烯醇（3Ac-DON）、15-乙酰脱氧雪腐镰刀菌烯醇（15Ac-DON）、DON-3-葡萄糖苷（DON-3Glc）、玉米赤霉烯酮和伏马菌素-X。测试了不同的样品制备技术：固相萃取、QuEChERS、OASIS HLB 柱或免疫亲和柱固相萃取和 Mycosep 225 Trich 柱。测试的六种免疫亲和柱均未显示出对所有霉菌毒素的交叉反应性。令人惊讶的是，结果表明，如果免疫亲和柱结合了 3Ac-DON，则它们不会结合 15Ac-DON。最有效的样品制备是使用 Mycosep 225 Trich 柱进行净化。优化了色谱条件，以实现所有分析物（包括 3Ac-DON 和 15Ac-DON 异构体）的完全分离。验证结果表明，重复性和再现性的相对标准偏差在 4%至 24%之间变化。表观回收率在 92%至 97%之间，定量限描述了 1.30 至 50μg/kg 的范围。通过能力验证和参考物质分析评估，该方法的准确度令人满意。用开发的方法分析了 99 个饲料样品，结果表明在 85%的动物饲料中检测到 DON 和 DON-3Glc，浓度分别为 1.70 至 1709μg/kg。在所调查的饲料中，DON-3Glc 与 DON 的比值从低至 3%到高至 59%不等。
