Simultaneous analysis of mycotoxins in corn-based animal feed by quadrupole-Orbitrap LC-MS/MS

Significance of the topic

The contamination of animal feed by fungal mycotoxins poses serious threats to livestock health, productivity and food safety worldwide. Regulatory bodies in North America, Europe and Asia have set maximum tolerance levels for key mycotoxins in feed ingredients. Routine monitoring is essential for safe feed formulation and to minimize economic losses resulting from animal illness or product condemnation.

Study objectives and overview

This study evaluated the performance of an Orbitrap Exploris 120 high-resolution accurate-mass (HRAM) mass spectrometer coupled to a Vanquish Flex UHPLC system for the rapid, simultaneous analysis of 15 major mycotoxins in corn-based feed. Three full-scan HRAM workflows were compared: data-independent acquisition (DIA), data-dependent acquisition (ddMS²) and targeted MS² (tMS²). Key aims included assessing sensitivity, linearity, precision, recovery and mass accuracy under a simple QuEChERS extraction protocol.

Methodology and instrumentation

Sample preparation used a 70:30 methanol–water extraction with 0.1% formic acid in a QuEChERS format, followed by centrifugation and filtration. Analysis was performed on a Vanquish Flex UHPLC with a Hypersil GOLD aQ column (100 × 2.1 mm, 1.9 µm) using a water/methanol gradient with ammonium formate and acids. The Exploris 120 MS executed rapid positive/negative polarity switching, full-scan detection at 30 000 FWHM and MS² fragmentation at 15 000 or 30 000 FWHM depending on the workflow.

Main results and discussion

Chromatographic separation achieved good peak shape for early-eluting polar toxins and baseline resolution for less polar fumonisins. All 15 analytes displayed excellent linearity (R²>0.995) from 2–4000 ng/g and average recoveries between 80% and 120% across DIA, ddMS² and tMS² modes. Precision studies (n=5) at mid-calibration levels yielded %RSD<12% for all compounds. Mass accuracy remained stable (<1 ppm) across chromatographic peaks in both polarities, with ample scan density for confident quantitation and MS² confirmation. Analysis of certified reference material confirmed measured concentrations within 20% of nominal values for all toxins.

Benefits and practical applications

• High-throughput screening and quantitation of multiple mycotoxins in a single 13 min run.
• Excellent sensitivity down to regulatory limit levels (ng/g range) with simple sample cleanup.
• Reliable MS² confirmation against curated spectral libraries via TraceFinder and mzCloud tools.
• Time savings by combining positive/negative mode analysis in one injection.

Future trends and opportunities

Advances in HRAM instrumentation will enable larger multi-mycotoxin panels and lower detection limits across diverse feed and food matrices. Coupling enhanced spectral libraries and automated data processing will further streamline routine monitoring. Novel acquisition strategies and application of machine learning for spectral deconvolution may improve trace-level screening of emerging mycotoxins and co-occurring contaminants.

Conclusion

The Orbitrap Exploris 120 UHPLC-HRAM platform delivers robust, accurate and high-throughput analysis of complex animal feed matrices. The combined use of DIA, ddMS² and tMS² workflows provides versatile quantitation and confirmation capabilities. Fast polarity switching and simple QuEChERS extraction make this approach well suited for routine food and feed safety laboratories aiming to meet stringent regulatory requirements.

References

1. Bennett JW, Klich M. Mycotoxins. Clinical Microbiology Reviews. 2003;16(3):497–516.
2. Zain ME. Impact of mycotoxins on human and animals. Journal of the Saudi Chemical Society. 2011;15(2):129–144.
3. Global guidelines for mycotoxin tolerance levels in animal feed. Reports from Canadian Food Inspection Agency, US FDA, European Commission and Chinese regulatory documents.