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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
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Mycophenolic acid, ≥98%
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Aflatoxin G2
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Magnesium sulfate solution, for molecular biology, 1.00 M±0.04 M
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Magnesium sulfate, BioReagent, suitable for cell culture, suitable for insect cell culture
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Deoxynivalenol
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Magnesium sulfate, ≥99.99% trace metals basis
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Acetonitrile, ≥99.5%, ACS reagent
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Acetonitrile, anhydrous, 99.8%
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Hydrogen chloride, ReagentPlus®, ≥99%
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T-2 Toxin, from Fusarium sp., powder, ≥98% (HPLC)
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Zearalenone, fungal mycotoxin
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Fumonisin B1 from Fusarium moniliforme, ≥98% (HPLC)
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Aflatoxin G1, from Aspergillus flavus
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Methanol-12C, 99.95 atom % 12C
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Magnesium sulfate solution, BioUltra, for molecular biology
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Magnesium sulfate, puriss. p.a., drying agent, anhydrous, ≥98.0% (KT), powder (very fine)
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Methanol solution, (Methanol:Dimethyl sulfoxide 1:1 (v/v))
Supelco
Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%
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Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
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Acetonitrile, HPLC Plus, ≥99.9%, poly-coated bottles
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Acetonitrile, ≥99.5% (GC)
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Magnesium sulfate, anhydrous, ReagentPlus®, ≥99.5%
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Acetonitrile, ReagentPlus®, 99%
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Magnesium sulfate, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99.5%
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Magnesium sulfate, anhydrous, reagent grade, ≥97%
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Magnesium sulfate, anhydrous, free-flowing, Redi-Dri, reagent grade, ≥97%
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Acetonitrile, suitable for DNA synthesis, ≥99.9% (GC)
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Acetonitrile, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.5% (GC)