Assessment of Dried Blood Spots for Multi-Mycotoxin Biomarker Analysis in Pigs and Broiler Chickens

• Published in: Toxins Vol. 11; no. 9; p. 541
• Main Authors: Lauwers, Marianne, Croubels, Siska, De Baere, Siegrid, Sevastiyanova, Milena, Romera Sierra, Eva Maria, Letor, Ben, Gougoulias, Christos, Devreese, Mathias
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 18.09.2019; MDPI
• Subjects: Aflatoxins; Beauvericin; Biomarkers; biomarkers for exposure; Blood; Blood volume; broiler chickens; Chickens; Chromatography; Coffee; dried blood spots; Enniatins; exposure assessment; Feeds; Filter paper; Fusarium; Hematocrit; Hogs; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Metabolites; multi-mycotoxin analysis; Mycotoxins; Ochratoxin A; pigs; Plasma; Poultry; Proteins; Sampling; Sampling methods; Scientific imaging; Solvents; Spectroscopy; toxicokinetic study; pigs; multi-mycotoxin analysis; toxicokinetic study; exposure assessment; biomarkers for exposure; broiler chickens; dried blood spots
• ISSN: 2072-6651; 2072-6651
• DOI: 10.3390/toxins11090541

Dried blood spots (DBSs), a micro-sampling technique whereby a drop of blood is collected on filter paper has multiple advantages over conventional blood sampling regarding the sampling itself, as well as transportation and storage. This is the first paper describing the development and validation of a method for the determination of 23 mycotoxins and phase I metabolites in DBSs from pigs and broiler chickens using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The targeted mycotoxins belong to groups for which the occurrence in feed is regulated by the European Union, namely, aflatoxins, ochratoxin A and several mycotoxins, and to two groups of unregulated mycotoxins, namely mycotoxins and mycotoxins (enniatins and beauvericin). The impact of blood haematocrit, DBS sampling volume and size of the analysed DBS disk on the validation results was assessed. No effects of variation in size of the analysed disk, haematocrit and spotted blood volume were observed for most mycotoxins, except for the aflatoxins and β-zearalanol (BZAL) at the lowest haematocrit (26%) level and for the enniatins (ENNs) at the lowest volume (40 µL). The developed method was transferred to an LC-high resolution mass spectrometry instrument to determine phase II metabolites. Then, the DBS technique was applied in a proof-of-concept toxicokinetic study including a comparison with LC-MS/MS data from plasma obtained with conventional venous blood sampling. A strong correlation ( > 0.947) was observed between plasma and DBS concentrations. Finally, DBSs were also applied in a pilot exposure assessment study to test their applicability under field conditions.