Determination of Fusarium Mycotoxins in Wheat, Maize and Animal Feed Using an Online TurboFlow and Orbitrap LC/MS Method

Significance of the Topic

Fusarium mycotoxins such as deoxynivalenol, zearalenone, T-2 toxin, HT-2 toxin and fumonisins pose a persistent threat to food and feed safety due to their stability and toxicity. Efficient and reliable analytical methods are essential to ensure compliance with legislative limits and protect human and animal health.

Study Objectives and Overview

This work presents a fully automated, online sample cleanup and high-resolution mass spectrometric method for simultaneous determination of six regulated Fusarium mycotoxins in wheat, maize and animal feed. The method aimed to minimize manual preparation, achieve high throughput, and deliver the sensitivity and accuracy required by EU regulations.

Methodology

• Sample preparation: Weigh 5 g of ground sample, extract with water/ACN (43:57) containing 0.1 % formic acid for 45 min, then filter through 0.2 µm nylon membrane.
• Online cleanup: Inject 10 µL of extract into the Transcend TLX-1 system equipped with a TurboFlow Cyclone MCX column for automatic matrix removal.
• Chromatography: Transfer analytes to a Hypersil GOLD analytical column (50 × 4.6 mm, 5 µm) with an 18 min gradient using water and methanol (both 0.1 % FA).
• Detection: Perform full-scan high-resolution Orbitrap MS (100 000 FWHM) in positive/negative switching mode (m/z 100–900) with HCD fragmentation for additional confirmation.
• Quantification: Use external matrix-matched calibration (8 levels) and peak-area comparison to determine concentrations.

Instrumentation

• Transcend TLX-1 online chromatography system with TurboFlow technology
• Exactive Orbitrap mass spectrometer
• Hypersil GOLD analytical column, 50 × 4.6 mm, 5 µm
• TurboFlow Cyclone MCX column, 0.5 × 50 mm
• CTC autosampler with 100 µL syringe, column oven (40 °C)

Main Results and Discussion

The method demonstrated excellent linearity (R² > 0.985) over regulatory concentration ranges. Limits of quantification in matrices ranged from 3 µg/kg (zearalenone) to 50 µg/kg (deoxynivalenol). Recoveries across wheat, maize and feed fell between 71.6 % and 120.2 % with intra- and inter-day precision (RSD) below 19 %. Mass accuracy remained under 2 ppm, and retention time shifts were within 2.5 %. Successful analysis of certified reference materials and proficiency testing confirmed accuracy and robustness.

Benefits and Practical Applications

The online TurboFlow cleanup eliminates labor-intensive immunoaffinity steps, reduces solvent use and instrument maintenance, and enables the analysis of over 200 samples without column replacement. The method is ideally suited for routine monitoring in food safety laboratories to ensure compliance with EU directives.

Future Trends and Applications

Advances may include expansion to additional mycotoxins, integration with automated data processing and real-time reporting, and coupling with other high-resolution platforms. Adapting the workflow to multi-residue screening will further enhance surveillance capabilities and risk assessment in food and feed matrices.

Conclusion

This study establishes a fast, sensitive and reliable online TurboFlow–Orbitrap LC-MS method for regulated Fusarium mycotoxins in complex matrices. It meets legislative requirements, streamlines laboratory workflows and offers high throughput with minimal maintenance.

Reference

• Commission Recommendation 576/2006 on presence of Fusarium toxins in animal feed
• Commission Decision 657/2002 on analytical method performance
• Regulation 1126/2007 amending EU limits for Fusarium toxins in maize
• Regulation 1881/2006 setting maximum levels for food contaminants
• Regulation 401/2006 on sampling and analysis methods for mycotoxins