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  • Development and validation of an UHPLC-MS/MS method for the determination of mycotoxins in grass silages.

Development and validation of an UHPLC-MS/MS method for the determination of mycotoxins in grass silages.

Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment (2015-09-17)
Cormac McElhinney, Pádraig O'Kiely, Chris Elliott, Martin Danaher
ABSTRACT

An ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) multi-mycotoxin analytical method was developed to simultaneously identify and quantify 20 mycotoxins in grass silages, inclusive of mycotoxins that are currently regulated in European Union feeds. Extraction of mycotoxins from dried grass silages was performed using of a modified QuEChERS extraction employing an acidified aqueous extraction (0.1 N HCl) with no further clean-up. Following chromatographic separation, analytes were detected using a fast polarity-switching MS/MS method that allowed both positive and negative ions to be analysed from a single injection, thus the reducing time and cost of analysis. The limits of detection and quantification ranged between 3 µg kg(-1) DM (aflatoxin B1, beauvericin and enniatin A and A1) and 200 µg kg(-1) DM (deoxynivalenol), and between 10 µg kg(-1) DM (aflatoxin B1, beauvericin and enniatin A1) and 500 µg kg(-1) DM (deoxynivalenol), respectively. Inter-assay accuracy and precision ranged between 90% and 107% and between 3.9% and 15.0% CV, respectively. The accuracy of the method was assessed through the application to a range of incurred samples in an inter-laboratory study.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Beauvericin, ≥97% (HPLC)
Sigma-Aldrich
Mycophenolic acid, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Mycophenolic acid, ≥98%
Sigma-Aldrich
Magnesium sulfate solution, for molecular biology, 1.00 M±0.04 M
Sigma-Aldrich
Magnesium sulfate, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Deoxynivalenol
Sigma-Aldrich
5α-Androstan-17β-ol-3-one, purum, ≥99.0% (TLC)
Sigma-Aldrich
Magnesium sulfate, ≥99.99% trace metals basis
Sigma-Aldrich
Acetonitrile, anhydrous, 99.8%
Sigma-Aldrich
Zearalenone, fungal mycotoxin
Sigma-Aldrich
Fumonisin B1 from Fusarium moniliforme, ≥98% (HPLC)
Sigma-Aldrich
Methanol-12C, 99.95 atom % 12C
Sigma-Aldrich
Magnesium sulfate solution, BioUltra, for molecular biology
Sigma-Aldrich
Magnesium sulfate, puriss. p.a., drying agent, anhydrous, ≥98.0% (KT), powder (very fine)
Sigma-Aldrich
Methanol solution, (Methanol:Dimethyl sulfoxide 1:1 (v/v))
Sigma-Aldrich
Magnesium sulfate, Vetec, reagent grade
Sigma-Aldrich
Acetonitrile, Preparateur, ≥99.9% (GC), One-time steel-plastic (SP) drum
Sigma-Aldrich
Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
Sigma-Aldrich
Ultrapure Acetonitrile
Sigma-Aldrich
Acetonitrile, ≥99.5% (GC)
Sigma-Aldrich
Magnesium sulfate, anhydrous, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Acetonitrile, ReagentPlus®, 99%
Sigma-Aldrich
Magnesium sulfate, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Magnesium sulfate, anhydrous, reagent grade, ≥97%
Sigma-Aldrich
Magnesium sulfate, anhydrous, free-flowing, Redi-Dri, reagent grade, ≥97%
Sigma-Aldrich
Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)
Sigma-Aldrich
Acetonitrile, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.5% (GC)
Sigma-Aldrich
Acetonitrile, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetonitrile, biotech. grade, ≥99.93%
Sigma-Aldrich
Acetonitrile, electronic grade, 99.999% trace metals basis