Multi-residue analysis of 18 regulated mycotoxins by LC-MS/MS

Significance of the Topic

Fusarium mycotoxins present a significant health risk in food and feed worldwide. Regulatory limits are enforced to protect human and animal health. Robust multi-residue methods enable efficient monitoring and ensure compliance with safety standards.

Objectives and Study Overview

This work aimed to develop and validate a single LC-MS/MS protocol for the simultaneous analysis of 18 regulated mycotoxins, including aflatoxins B1 B2 G1 G2, fumonisins B1 B2 B3, ochratoxin A and a range of trichothecenes. Key goals were low limits of quantification at or below EC 1881 2006 requirements, separation of critical regioisomers and reliable performance across complex matrices.

Methodology and Instrumentation

• Sample extracts prepared by validated protocols at Concept Life Sciences.
• Chromatography on Shimadzu Nexera UHPLC with Mastro PFP 100 x 2.1 mm 3 µm column at 40 °C and 0.4 mL/min flow.
• Mobile phases: 0.15 mM ammonium fluoride in water (A) and 0.15 mM ammonium fluoride with 2 % acetic acid in methanol (B).
• Detection using Shimadzu LCMS-8060 triple quadrupole with positive/negative polarity switching.
• Quantification via 13C labelled internal standards spanning the 18 analytes.

Used Instrumentation

• Shimadzu Nexera UHPLC system
• Shimadzu LCMS-8060 triple quadrupole mass spectrometer
• Mastro PFP bonded phase column (100 x 2.1 mm, 3 µm)

Main Results and Discussion

Use of ammonium fluoride significantly enhanced positive-ion signal for all targets compared with ammonium acetate, formate and acetic acid systems. The PFP column achieved near baseline separation of challenging isomer pairs such as 3-AcDON/15-AcDON and FB2/FB3. Limits of quantification met or exceeded legislative values. Analyses of mixed spice and pepper extracts spiked with target mycotoxins yielded repeatability below 10 % RSD over 12 injections.

Benefits and Practical Applications

• High throughput with a 12.5 minute analysis cycle.
• Simultaneous quantification of 18 mycotoxins reduces labor and sample consumption.
• Enhanced sensitivity supports trace detection for regulatory compliance.
• Robust performance in diverse food matrices enables routine QA/QC applications.

Future Trends and Opportunities

Future developments may include extension to emerging mycotoxins, coupling with high-resolution mass spectrometry for improved selectivity, automation of extraction protocols, AI-assisted data processing and deployment of portable LC-MS platforms for on-site screening.

Conclusion

The developed LC-MS/MS method offers a robust, sensitive and efficient solution for multi-class mycotoxin monitoring. Its capability in isomer separation, regulatory compliance and matrix robustness supports widespread adoption in analytical laboratories.

References

• Baker D Titman C Loftus N Horner J Multi residue analysis of 18 regulated mycotoxins by LC MS MS Shimadzu Manchester and Concept Life Sciences Cambridge