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

Significance of the Topic

Cereals are staples in global diets and feed chains, yet they can be contaminated by a wide array of mycotoxins and plant toxins such as tropane alkaloids. These compounds threaten food safety, public health, and regulatory compliance. Rapid, sensitive, and comprehensive analytical methods are essential to monitor both regulated and emerging toxins, support risk assessment, and guide mitigation strategies in food and feed production.

Objectives and Study Overview

This study aimed to extend a previously developed UPLC-MS/MS method to quantify 50 regulated and emerging mycotoxins, plus atropine and scopolamine, in cereal-based commodities. The focus was on achieving low detection limits, broad quantitation ranges, and compliance with SANTE guidelines, while simplifying sample preparation.

Methodology and Instrumentation

Sample preparation relied on a dilute-and-shoot protocol without cleanup or internal standards. A flour blend (wheat, barley, rice, maize) was extracted with acidified acetonitrile–water, diluted tenfold, and filtered prior to injection. Chromatographic separation used an ACQUITY UPLC I-Class PLUS system with a BEH C18 column at 40 °C. Mobile phases consisted of ammonium acetate in water and acidified methanol. The Xevo TQ-XS tandem quadrupole mass spectrometer operated in ESI positive/negative mode with polarity switching in multiple reaction monitoring (MRM). Data acquisition was via MassLynx v4.2 and quantitation performed with waters_connect MS Quan.

Key Results and Discussion

• Instrument limits of detection were in the sub-ng/mL range (e.g. 0.0003 ng/mL for aflatoxins).
• Method limits of quantification met regulatory thresholds: 0.1 µg/kg for aflatoxins; ≤1 µg/kg for major toxins; ≤10 µg/kg for most analytes.
• Calibration curves showed excellent linearity (R2 ≥ 0.99) over three orders of magnitude.
• Matrix effects varied widely (–100% to +83%), highlighting the need for matrix-matched calibration or labeled internal standards.
• Repeatability was strong, with RSDr ≤10% and retention time stability within ±0.03 min.

Practical Implications and Benefits

The combined UPLC separation and Xevo TQ-XS sensitivity allow high sample dilution, reducing matrix interferences and simplifying workflows. A quick dilute-and-shoot approach can replace extensive cleanup, increasing throughput. The method fulfills regulatory criteria for mycotoxin control and can be applied by food and feed laboratories for compliance testing and exposure assessment.

Instrumentation

• UPLC: ACQUITY UPLC I-Class PLUS with BEH C18 column
• MS/MS: Xevo TQ-XS triple quadrupole
• Software: MassLynx v4.2 and waters_connect MS Quan application

Future Trends and Applications

Advances in high-resolution chromatography and mass spectrometry will enable inclusion of additional emerging and modified mycotoxins. Integration of isotope-labeled standards will further improve quantitation accuracy. Automated data processing platforms like waters_connect will accelerate reporting. The method framework can be adapted to other complex matrices, and ongoing surveillance of new toxin metabolites will strengthen food safety programmes.

Conclusion

This multi-toxin UPLC-MS/MS method delivers high sensitivity, broad coverage, and robust quantitation for 50 mycotoxins and tropane alkaloids in cereals. Its simplicity and compliance with international guidelines make it a valuable tool for routine monitoring and risk management in the food and feed industries.

References

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5. Dreolin N. and Stead S. Waters Application Note 720006685EN, 2021.
6. Dreolin N. et al. Waters Application Note 720007165EN, 2021.
7. SANTE 12089/2016. Guidance Document on Identification of Mycotoxins in Food and Feed.