Multi-mycotoxin Analysis in Corn-Based Feed by Quadrupole-Orbitrap LC-MS/MS

Importance of the Topic

Mycotoxins in animal feed pose significant health risks and economic losses worldwide. Rapid and accurate detection of multiple toxins in complex matrices such as corn-based feed is essential for ensuring animal safety, meeting regulatory limits, and minimizing contaminated batches in the supply chain.

Objectives and Study Overview

This study aimed to evaluate the performance of a Thermo Scientific™ Orbitrap Exploris™ 120 high-resolution mass spectrometer coupled to ultra-high-pressure liquid chromatography (UHPLC) for simultaneous identification and quantitation of 15 priority mycotoxins in corn feed. Three MS acquisition workflows—data independent acquisition (DIA), data dependent acquisition (DDA), and targeted MS2 (tMS2/PRM)—were compared for robustness, accuracy, and throughput.

Methodology and Instrumentation

The analytical protocol combined a modified QuEChERS extraction of 5 g feed samples with injection of 2 µL into a UHPLC–MS system. Key steps included:

• Extraction with 70:30 methanol-water (0.1% formic acid)
• 35 min vortex mixing and centrifugation
• 0.45 µm filtration prior to injection

MS data acquisition employed rapid polarity switching for full-scan MS and MS/MS across 100–1000 m/z. TraceFinder™ software with mzVault™ and mzCloud™ libraries was used for automated peak integration, fragment matching, and confirmation.

Instrumentation

• UHPLC: Thermo Scientific™ Vanquish™ Flex Binary System
• Column: Hypersil GOLD aQ, 100×2.1 mm, 1.9 µm
• Mobile phases: Water and methanol, each with 5 mM ammonium formate, 0.1% formic and acetic acids
• Mass spectrometer: Thermo Scientific™ Orbitrap Exploris™ 120 with OptaMax™ NG ion source

Main Results and Discussion

Chromatographic conditions achieved clear separation of all 15 compounds, including early-eluting trichothecenes and later-eluting fumonisins. Key findings:

• Excellent peak shape and retention stability over 13 min runs
• Linearity (r² > 0.995) across 2–100 ng/g with recoveries of 80–120%
• Mass accuracy within ±5 ppm for precursor and fragment ions under polarity switching
• Comparable quantitative performance across DIA, DDA, and tMS2 modes
• Certified reference material analysis confirmed accuracy within 20% of nominal values

Benefits and Practical Applications

The workflow offers high confidence in screening and quantitation of multiple mycotoxins in a single injection. Its rapid polarity switching, full-scan capability, and integrated spectral libraries support routine feed quality control, regulatory compliance, and research on toxin co-occurrence.

Future Trends and Potential Applications

Advances may include:

• Expansion of spectral libraries to emerging toxins
• Integration of machine learning for automated peak annotation
• Miniaturized and field-deployable high-resolution MS for on-site testing
• Real-time monitoring workflows in production environments

Conclusion

The Orbitrap Exploris 120 UHPLC–MS/MS platform, combined with optimized sample preparation and acquisition strategies, delivers precise, accurate, and high-throughput multi-mycotoxin analysis in corn-based feed. All three scan modes provided reliable quantitation and confirmation, making this approach fit-for-purpose in QA/QC and research laboratories.

Reference

1. Bennett JW, Klich M. Mycotoxins. Clin Microbiol Rev. 2003;16:497–516.
2. Zain ME. Impact of mycotoxins on human and animals. J Saudi Chem Soc. 2011;15:129–144.