Landeslabor Berlin-Brandenburg, Rudower Chaussee 39, D-12489 Berlin, Germany.
Eurofins Sofia GmbH, Rudower Chaussee 29, D-12489 Berlin, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jun 1;1117:86-102. doi: 10.1016/j.jchromb.2019.04.013. Epub 2019 Apr 6.
A 2D LC-MS/MS method for the simultaneous determination of 350 pesticides, 16 mycotoxins as well as the growth regulators Chlormequat and Mepiquat was developed. The method is applicable to cereals and products thereof. Attention should be paid to the simultaneous analysis of the cereal-relevant mycotoxins aflatoxin B1, B2, G1 and G2, ochratoxin A, deoxynivalenol and zearalenone. Moreover, the tropane alkaloids atropine/scopolamine could be integrated into the final method. The samples were extracted with a mixture of acetonitrile/water (80:20), diluted with acetonitrile and injected into an LC-LC-MS/MS system. There were no further manual clean-up steps. The automatic online clean-up took place during the HILIC-separation in the first dimension (YMC-Pack Diol; 2.1 × 100 mm; 5 μm, 120 Å). Here, polar matrix compounds were retained, while the majority of the analyte scope eluted in a fraction at the beginning of the analytical run. This fraction was transferred to the second dimension by a packed loop interface (Agilent Zorbax SB-C8; 4.6 × 12.5 mm; 5 μm; 80 Å). On the second column (Phenomenex Synergi Fusion RP C18; 2 × 100 mm; 2.5 μm; 100 Å), the majority of the scope was separated by a typical RP-gradient. Only some of the polar pesticides could not be transferred to the second column. They eluted directly after the transfer step from the HILIC-column to the MS/MS. The final method was sensitive enough to meet all the regulated maximum levels for pesticides in cereals according to EU Regulation 396/2005 and those for contaminants according to EU Regulation 1881/2006. Above all, the method was so robust and accurate that nearly 90% of the pesticides and all the tested mycotoxins, growth regulators and tropane alkaloids fulfilled the validation criteria of the SANTE guideline document, although the demanding criteria are only applicable to pesticides. For the verification, eight proficiency tests were passed successfully: three for the pesticide analysis, three for the mycotoxin analysis, and two for the analysis of the tropane alkaloids. In addition to the already mentioned contaminants, the six most important ergot alkaloids (e.g. ergotamine/ergotaminine) and two modified mycotoxins (deoxynivalenol-3-glucoside and zearalenone-sulfate, also known as masked mycotoxins) were detected during the routine analysis of rye and corn samples.
开发了一种用于同时测定 350 种农药、16 种真菌毒素以及生长调节剂矮壮素和甲哌𬭩的 2D LC-MS/MS 方法。该方法适用于谷物及其制品。需要注意的是，同时分析与谷物相关的真菌毒素黄曲霉毒素 B1、B2、G1 和 G2、赭曲霉毒素 A、脱氧雪腐镰刀菌烯醇和玉米赤霉烯酮。此外，托烷生物碱阿托品/东莨菪碱也可以整合到最终方法中。样品用乙腈/水（80:20）混合提取，用乙腈稀释后注入 LC-LC-MS/MS 系统。无需进一步手动净化步骤。在第一维（YMC-Pack Diol；2.1×100mm；5μm，120Å）的 HILIC 分离过程中自动进行在线净化。在这里，极性基质化合物被保留，而大多数分析物范围在分析运行开始时的一个馏分中洗脱。该馏分通过填充环接口（Agilent Zorbax SB-C8；4.6×12.5mm；5μm；80Å）转移到第二维。在第二根柱子（Phenomenex Synergi Fusion RP C18；2×100mm；2.5μm；100Å）上，通过典型的 RP 梯度分离大部分范围。只有一些极性农药不能转移到第二根柱子上。它们在从 HILIC 柱转移到 MS/MS 后直接洗脱。最终方法足够灵敏，能够满足根据欧盟法规 396/2005 对谷物中农药的所有规定最大水平以及根据欧盟法规 1881/2006 对污染物的规定最大水平。最重要的是，该方法非常稳健和准确，尽管苛刻的标准仅适用于农药，但近 90%的农药和所有测试的真菌毒素、生长调节剂和托烷生物碱都符合 SANTE 指导文件的验证标准。为了验证，成功通过了八项能力验证测试：三项用于农药分析，三项用于真菌毒素分析，两项用于托烷生物碱分析。除了已经提到的污染物外，在黑麦和玉米样品的常规分析中还检测到六种最重要的麦角生物碱（如麦角胺/麦角新碱）和两种修饰真菌毒素（脱氧雪腐镰刀菌烯醇-3-葡萄糖苷和玉米赤霉烯酮-硫酸盐，也称为掩蔽真菌毒素）。
