Simultaneous Determination of Alternaria Toxins, Ergot Alkaloid Epimers and other major Mycotoxins in various Food Matrices by LC-MS/MS

Importance of the Topic

Food safety monitoring demands sensitive, reliable methods to detect mycotoxins that compromise human and animal health. Alternaria toxins and ergot alkaloid epimers frequently co-occur with major mycotoxins in cereal-based products, yet conventional protocols analyze these groups separately. A unified LC-MS/MS approach delivers comprehensive surveillance in a single, streamlined workflow.

Study Objectives and Overview

This study aimed to develop and validate a single LC-MS/MS method for the simultaneous quantification of 37 target analytes, including Alternaria toxins, ergot alkaloid epimers and key mycotoxins (aflatoxins, fumonisins, trichothecenes, zearalenone, ochratoxin A, citrinin, patulin). Validation across diverse food matrices ensures method applicability for routine testing and regulatory compliance.

Methodology and Instrumentation

A single-step extraction protocol (0.5% formic acid in 80/20 acetonitrile/water) was optimized for baby cereal, peanut, tomato puree and blended flour to minimize matrix effects and maximize recovery. Chromatographic separation employed a Raptor Biphenyl column (2.7 µm, 100 × 2.1 mm) with a guard cartridge at 60 °C, using 0.05% formic acid in water (A) and in methanol (B) under a 25–50–100–25% B gradient at 0.4 mL/min. Positive electrospray ionization MS/MS detection achieved baseline separation of ergot epimers and clear integration of polar Alternaria toxins.

Instrumentation Used:

• Ultra-high-performance LC system with Raptor Biphenyl analytical and guard columns
• Mobile phases: 0.05% formic acid in water and methanol
• MS/MS platform in positive ESI mode (e.g., Waters Acquity TQS or Shimadzu Nexera X2-8060)

Key Results and Discussion

Calibration curves exhibited r2 > 0.997 with deviations <20%. Limits of quantification reached 0.4 µg/kg for most analytes. Average recoveries ranged 72–112% (RSD ≤ 12%) across matrices, except citrinin in solids (26–35%). Baby cereal showed matrix interference blocking nivalenol detection. Chromatograms confirmed effective separation of ergot alkaloid epimers and reliable quantification of tenuazonic acid on the biphenyl phase.

Benefits and Practical Applications

This workflow enables:

• Comprehensive multi-mycotoxin screening in a single run
• Rapid sample preparation without extensive cleanup
• High throughput suitable for quality control and regulatory labs
• Robust performance across fat-, water- and carbohydrate-rich foods

Future Trends and Opportunities

Advances may include automated online extraction, expansion to emerging toxins, integration with high-resolution MS for non-target screening and application of machine learning for spectral interpretation. Real-time monitoring platforms and miniaturized devices could further accelerate food safety surveillance.

Conclusion

The presented LC-MS/MS method achieves unique simultaneous detection of Alternaria toxins, ergot alkaloid epimers and major mycotoxins with excellent sensitivity, accuracy and precision. Its simple workflow and broad matrix suitability offer a powerful tool for modern food contaminant analysis.