Multi LC-MS/MS and LC-HRMS Methods for Determination of 24 Mycotoxins including Major Phase I and II Biomarker Metabolites in Biological Matrices from Pigs and Broiler Chickens
Multi LC-MS/MS and LC-HRMS Methods for Determination of 24 Mycotoxins including Major Phase I and II Biomarker Metabolites in Biological Matrices from Pigs and Broiler Chickens
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DOI：
10.3390/toxins11030171
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发表时间：
2019-03-19
期刊：
影响因子：
4.2
通讯作者：
Devreese, Mathias
中科院分区：
文献类型：
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作者：
Lauwers, Marianne;De Baere, Siegrid;Devreese, Mathias
建立了一种可靠、实用的检测猪血浆、尿液和粪便、肉鸡血浆和排泄物中真菌毒素的高效液相色谱-串联质谱法。目标真菌毒素属于受监管的类群，即黄曲霉毒素、赭曲霉毒素和镰刀菌毒素，以及两类新出现的真菌毒素，即交链孢霉毒素和冬虫夏草毒素。此外，所开发的方法被转移到LC-高分辨率质谱仪，以定性地测定I期和II期代谢物，这些代谢物的分析标准并不总是商业上可用的。血浆的样品制备简单而通用，通过单独沉淀蛋白质(猪)或结合去除磷脂(鸡)来完成。需要对其他基质进行更密集的样品净化，包括使用乙酸乙酯(猪尿)、甲醇/乙酸乙酯/甲酸(75/24/1，v/v/v)(猪粪便)或乙腈(鸡粪)的依赖pH的液液萃取(LLE)。此外，对于猪粪便的提取，使用丙酮的LLE和上清液在Hybridge SPE-磷脂小柱上的过滤相结合。LC-MS/MS方法根据欧洲和国际社会制定的指南进行了内部验证。最后，多种方法被成功地应用于特定的毒代动力学研究和筛查研究，以监测单个动物的暴露。
A reliable and practical multi-method was developed for the quantification of mycotoxins in plasma, urine, and feces of pigs, and plasma and excreta of broiler chickens using liquid chromatography-tandem mass spectrometry. The targeted mycotoxins belong to the regulated groups, i.e., aflatoxins, ochratoxin A and Fusarium mycotoxins, and to two groups of emerging mycotoxins, i.e., Alternaria mycotoxins and enniatins. In addition, the developed method was transferred to a LC-high resolution mass spectrometry instrument to qualitatively determine phase I and II metabolites, for which analytical standards are not always commercially available. Sample preparation of plasma was simple and generic and was accomplished by precipitation of proteins alone (pig) or in combination with removal of phospholipids (chicken). A more intensive sample clean-up of the other matrices was needed and consisted of a pH-dependent liquid-liquid extraction (LLE) using ethyl acetate (pig urine), methanol/ethyl acetate/formic acid (75/24/1, v/v/v) (pig feces) or acetonitrile (chicken excreta). For the extraction of pig feces, additionally a combination of LLE using acetone and filtration of the supernatant on a HybridSPE-phospholipid cartridge was applied. The LC-MS/MS method was in-house validated according to guidelines defined by the European and international community. Finally, the multi-methods were successfully applied in a specific toxicokinetic study and a screening study to monitor the exposure of individual animals.