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Open AccessArticle

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

1
Department of Pharmacology, Toxicology and Biochemistry, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium
2
Innovad, Postbaan 69, 2910 Essen, Belgium
3
Chair of Analytical Food Chemistry, Technische Universität München, Maximus-von-Imhof-Forum 2, 85354 Freising, Germany
*
Author to whom correspondence should be addressed.
Toxins 2019, 11(3), 171; https://doi.org/10.3390/toxins11030171
Received: 15 January 2019 / Revised: 1 March 2019 / Accepted: 12 March 2019 / Published: 19 March 2019
(This article belongs to the Special Issue Application of LC-MS/MS in the Mycotoxins Studies)
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. View Full-Text
Keywords: Biomarkers; exposure; LC-MS/MS; LC-HRMS; pig; broiler chicken; multi-mycotoxin Biomarkers; exposure; LC-MS/MS; LC-HRMS; pig; broiler chicken; multi-mycotoxin
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Lauwers, M.; De Baere, S.; Letor, B.; Rychlik, M.; Croubels, S.; Devreese, M. 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. Toxins 2019, 11, 171.

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