Solutions for Flexible Work Styles LCMS Food Safety Applications

Significance of the topic

Mycotoxins produced by Fusarium species pose serious health risks to humans and animals through contaminated food and feed. To protect public health, maximum residue limits are set by the EU and other regulatory bodies for key mycotoxins such as aflatoxins, fumonisins, ochratoxin A and trichothecenes. Rapid, reliable multi-residue methods are required to monitor these compounds at low concentrations in complex matrices.

Objectives and study overview

This study aimed to develop and validate a single, high-throughput LC-MS/MS method capable of simultaneously quantifying eighteen regulated mycotoxins in food samples. The targeted analytes included aflatoxins B1, B2, G1, G2; fumonisins B1-B3; ochratoxin A; and trichothecenes such as deoxynivalenol, 3- and 15-acetyldeoxynivalenol, diacetoxyscirpenol, fusarenon X, HT-2, T-2, neosolaniol, nivalenol and zearalenone.

Methodology and instrumentation

The method used a Nexera X2 UHPLC coupled to a LCMS-8060 triple quadrupole MS operated in ESI positive/negative modes. Separation was achieved on a pentafluorophenyl (PFP) column using a gradient of methanol and aqueous ammonium fluoride (0.15 mM) to enhance ionization. Chromatographic cycle time was 12.5 minutes. Quantitation employed 2–3 MRM transitions per analyte, with 13C-labeled internal standards for calibration. Sample extracts were provided by Concept Life Sciences following validated QuEChERS-based protocols.

Main results and discussion

• Chromatographic resolution: The PFP column baseline-resolved regioisomers (3-/15-AcDON and FB2/FB3), which coelute on C18.
• Sensitivity: Limits of quantitation (LOQ) met EU requirements (0.1–10 µg/kg for aflatoxins; 10–1000 µg/kg for fumonisins and trichothecenes).
• Linearity: Calibration curves were linear across required ranges with R2>0.998.
• Repeatability: Intra-day RSDs were <10% for all analytes.
• Matrix performance: No appreciable matrix effect was observed in spiked wheat samples after cleanup, with recoveries of 90–107% for all toxins.

Benefits and practical applications of the method

• Comprehensive coverage: Simultaneous analysis of 18 mycotoxins in a single run.
• High throughput: 12.5 min cycle time enables large sample batches.
• Robust quantitation: Excellent linearity and repeatability without matrix-matched calibration.
• Enhanced selectivity: PFP column and ammonium fluoride improve separation and sensitivity.
• Regulatory compliance: Meets EU Regulation (EC) 1881/2006 requirements for mycotoxin monitoring.

Future trends and potential uses

• Method expansion: Extend to emerging mycotoxins and masked metabolites.
• Automation: Integrate online cleanup and sample preparation robotics.
• HRMS integration: Combine with high-resolution MS libraries for untargeted screening.
• Global harmonization: Standardize multi-residue mycotoxin methods across laboratories.

Conclusion

A single LC-MS/MS method using a PFP column and ammonium fluoride additive enables fast, sensitive and reliable multi-residue analysis of 18 regulated mycotoxins. The approach achieves required regulatory limits, high throughput and robust performance, making it suitable for routine food safety monitoring.

References

• European Commission, Regulation (EC) No 1881/2006; amending Regulation (EC) No 1126/2007.
• European Commission, Regulation (EC) No 401/2006 on sample preparation for mycotoxin determination.
• R. A. Rahmani et al., “Qualitative and Quantitative Analysis of Mycotoxins,” Compr. Rev. Food Sci. Food Saf., 2009.