Development of a Multi-Toxin UPLC-MS/MS Method for 50 Mycotoxins and Tropane Alkaloids in Cereal Commodities

Significance of the Topic

The monitoring of mycotoxins and tropane alkaloids in cereal-based foods is critical for ensuring food and feed safety globally. Regulatory limits for multiple toxins are enforced in many regions, and emerging or modified mycotoxins increasingly challenge conventional testing methods. A robust multi-toxin analytical method supports compliance, risk assessment, and public health protection.

Objectives and Study Overview

This work aimed to develop and validate a single UPLC-MS/MS method capable of detecting and quantifying 50 regulated and emerging mycotoxins, along with tropane alkaloids, in cereal commodities. Key goals included achieving low detection limits, broad calibration ranges, and compliance with SANTE guidelines using a simplified sample preparation approach.

Methodology and Instrumentation

• Sample Extraction: Dilute-and-shoot protocol with 80:20 acetonitrile:water (0.75% acetic acid, 0.2% formic acid), 1:10 dilution.
• Chromatography: ACQUITY UPLC I-Class PLUS, BEH C18 column, water and methanol mobile phases with ammonium acetate, acetic and formic acids.
• Mass Spectrometry: Xevo TQ-XS triple quadrupole in ESI+/- mode with polarity switching, MRM acquisition.
• Quantitation: Solvent and matrix-matched calibration curves; waters_connect with MS Quan for data processing.

Main Results and Discussion

• Method Limits of Quantification (m-LOQs) as low as 0.1 µg/kg for aflatoxins and ≤1 µg/kg for key toxins.
• Wide calibration range over three orders of magnitude with R2 ≥0.99 and residuals ≤20%.
• Significant matrix effects (−100% to +83%), highlighting the necessity of matrix-matched calibration.
• Repeatability with RSDr ≤10% and compliance with SANTE identification criteria.

Benefits and Practical Applications

• Simultaneous determination of over 50 toxins in a single run.
• Exceptional sensitivity and linearity fulfilling regulatory standards.
• Streamlined sample preparation reducing laboratory workload.
• Accelerated data review with waters_connect and MS Quan.

Future Trends and Potential Applications

As regulatory focus expands to include emerging and modified mycotoxins, this platform can be extended to new analytes. Incorporation of SPE clean-up, further dilution strategies, or labeled internal standards will enhance robustness. Advanced informatics tools will streamline high-throughput testing and data management.

Conclusion

The developed UPLC-MS/MS method on the Xevo TQ-XS and ACQUITY UPLC system delivers high sensitivity, precision, and compliance for multi-toxin analysis in cereals. Its simplicity and adaptability make it a valuable tool for routine and emerging toxin surveillance.

Reference

1. Eskola et al. Worldwide Contamination of Food-Crops With Mycotoxins: Validity of the ‘FAO Estimate’ of 25% in Crit. Rev. Food Sci. Nutr. 60(16):2773–2789 (2020).
2. Gruber-Dorninger et al. Emerging Mycotoxins: Beyond Traditionally Determined Food Contaminants in J. Agric. Food Chem. 65(33):7052–7070 (2017).
3. Freire & Sant’Ana. Modified Mycotoxins: Formation, Detection, Occurrence, and Toxic Effects in Food Chem. Toxicol. 111:189–205 (2018).
4. de Nijs et al. Short Inventory of EU Legislation on Plant Toxins in Food in World Mycotoxin J. 9:129–139 (2017).
5. Dreolin & Stead. Quantitative Determination of Regulated Mycotoxins in Cereal Flours on Xevo TQ-XS. Waters Application Note 720006685EN (2021).
6. Dreolin et al. Interlaboratory Study of Mycotoxins in Cereals by LC-MS/MS. Waters Application Note 720007165EN (2021).
7. SANTE 12089/2016. Guidance Document on Identification of Mycotoxins in Food and Feed.
8. Dreolin et al. Determination of Regulated and Emerging Mycotoxins in Foods Using SPE and UPLC-MS/MS. Waters Application Note 720007377EN (2021).