Determination of Regulated and Emerging Mycotoxins in Cereals, Nuts, Figs, and Animal Feeds Using Pass-Through SPE and UPLC- MS/MS

Importance of the Topic

Mycotoxin contamination in cereals, nuts, dried fruits and animal feeds poses serious health risks worldwide and is tightly regulated. A rapid, reliable multi-analyte method enhances food safety monitoring by detecting both regulated and emerging mycotoxins at low levels across diverse matrices.

Study Objectives and Overview

This work aimed to develop and validate a single LC-MS/MS method for 31 mycotoxins (including aflatoxins, ochratoxin A, fumonisins, trichothecenes, zearalenone, enniatins, Alternaria toxins and others) in wheat flour, tree and ground nuts, dried figs and animal feeds. Key goals were high recovery, regulatory compliance and streamlined sample cleanup.

Methodology and Used Instrumentation

A unified sample prep used acidified acetonitrile–water extraction followed by pass-through cleanup on Oasis PRiME HLB SPE cartridges. After loading and air-drying the cartridge, purified extract was diluted 1:5, filtered and injected.

Instrumentation:

• UPLC: ACQUITY UPLC I-Class PLUS with FTN injector
• Column: ACQUITY UPLC BEH C18, 2.1×100 mm, 1.7 µm
• Mass Spectrometer: Xevo TQ-S micro in ESI+/- with MRM
• Software: MassLynx v4.2 and TargetLynx XS

Main Results and Discussion

Chromatography delivered sharp peaks and baseline separation of critical pairs (e.g. DON vs. DON-3-Glu). Linearity (R²>0.99) and ion ratio criteria met SANTE guidelines. Instrument LOQs were ≤0.25 µg/kg for aflatoxins and ≤1 µg/kg for ochratoxin A. Method LOQs ranged from 0.25 to 10 µg/kg depending on matrix.

Spike recovery studies across all matrices yielded 60–108% with repeatability RSDs <11%. Analysis of six certified reference materials (maize flour, hazelnut slurry, fig slurry, feed) showed trueness 71–99% and RSD ≤7%.

Benefits and Practical Applications

• Single assay for 31 mycotoxins across multiple commodity types
• Rapid pass-through SPE eliminates conditioning and washing steps
• Regulatory-compliant sensitivity and precision
• Scalable to additional analytes and matrices

Future Trends and Opportunities

Regulations are expected to expand to include more modified and emerging mycotoxins. The broad applicability of Oasis PRiME HLB allows method extension to new targets. Integration with high-throughput workflows and advanced data processing will support routine monitoring under evolving legislative frameworks.

Conclusion

A robust, high-throughput UPLC-MS/MS method with simple SPE cleanup has been demonstrated for regulated and emerging mycotoxins in complex food and feed matrices. It meets current regulatory requirements, offers excellent accuracy and repeatability, and can be adapted to future analytical demands.

References

1. Agriopoulou et al. Foods 2020, 9(2):137.
2. Waters Oasis PRiME HLB Care & Use Manual.
3. Eurachem Guide 2nd ed. 2014.
4. SANTE/12089/2016 guidelines.
5. EU Regulation No 401/2006.