Multi-mycotoxin Screening and Quantitation Using UHPLC, High Resolution and Accurate Mass

Significance of the Topic

The presence of multiple mycotoxins in cereals and beer poses significant health risks. Fungi such as Fusarium, Aspergillus, Penicillium and Alternaria produce diverse toxic metabolites that can persist through malting and brewing. Rapid and reliable screening of these contaminants is essential for consumer safety and regulatory compliance.

Objectives and Overview of the Study

This work aimed to develop a streamlined method for simultaneous detection and quantitation of 32 mycotoxins in beer. The approach combined minimal sample handling with ultra high performance liquid chromatography and full-scan high resolution accurate mass spectrometry to enable both targeted analysis and retrospective data review.

Methodology and Instrumentation

Sample preparation involved degassing a beer aliquot, protein precipitation with acetonitrile, centrifugation, evaporation, reconstitution in methanol:water and microfiltration. Stable isotope labeled deoxynivalenol and zearalenone were added as surrogates. Chromatographic separation used a reversed-phase column with a water-ammonium formate and methanol gradient at 500 µL/min and 40 °C. Detection was performed on an Orbitrap mass spectrometer operated in full-scan APCI mode at resolution settings up to 100 000 FWHM.

Key Results and Discussion

APCI ionization improved sensitivity for most Fusarium toxins by up to 1200% compared with electrospray. Higher resolving power settings markedly enhanced signal clarity and selectivity, enabling detection of analytes at low microgram per liter levels. Matrix effects were assessed by comparing solvent and matrix standards; signal suppression or enhancement ranged from 63 to 112%. Limits of quantitation in matrix were established between 0.5 and 60 µg/L. Method validation showed recoveries from 92 to 124% and RSDs below 28% across spiking levels.

Benefits and Practical Applications

• Simplified sample handling allows high throughput testing.
• Full-scan data acquisition supports retrospective screening of additional compounds.
• High mass accuracy and resolution ensure reliable identification in complex beer matrices.
• Simultaneous quantitation of 32 mycotoxins streamlines quality control in brewing industry laboratories.

Future Trends and Potential Applications

Improvements may include integration of automated sample preparation, expansion to other beverages and grains, and coupling with advanced data mining tools. Further enhancements in resolution and ionization strategies could lower detection limits and broaden the range of detectable contaminants.

Conclusion

The developed UHPLC-Orbitrap APCI method offers a robust alternative to tandem MS for multi-mycotoxin analysis. It combines minimal sample preparation with high throughput, excellent mass accuracy and resolution, enabling reliable screening and quantitation of a broad spectrum of toxins in beer.

Reference

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