ANALYSIS OF MYCOTOXINS IN FOOD MATRICES - Agilent Ultivo Triple Quadrupole LC/MS System

Importance of the Topic

Mycotoxins are toxic secondary metabolites produced by fungi that frequently contaminate grains, nuts, spices and other foodstuffs. Their presence even at low levels poses serious health risks to humans and livestock. Regulatory agencies worldwide set strict maximum residue limits (MRLs) to prevent exposure. Achieving sensitive and accurate quantification of multiple mycotoxins across diverse food matrices is therefore essential for food safety monitoring and compliance.

Objectives and Study Overview

The primary goal of this study was to evaluate the performance of the Agilent Ultivo Triple Quadrupole LC/MS system for quantitative analysis of up to 12 regulated mycotoxins in three common food matrices: corn, peanut and black pepper. Key objectives included:

• Developing a single 9-minute dynamic multiple reaction monitoring (dMRM) method.
• Assessing method sensitivity down to fractions of the regulatory MRLs.
• Demonstrating precision, linearity and robustness across matrices.

Methodology and Instrumentation

Sample Preparation

• Corn and peanut: 5 g samples extracted with acetonitrile, water and enzyme-neutralizing salts, followed by dispersive solid-phase extraction (dSPE) cleanup with lipid removal sorbent.
• Black pepper: 2 g samples extracted similarly, cleaned with a universal dSPE kit and final extract diluted 30:70 with water.

Chromatography and Mass Spectrometry

• LC system: Agilent 1290 Infinity II (or 1260 Infinity II Prime) with an Eclipse Plus C18 column (3.0 × 150 mm, 1.8 µm) at 45 °C, injection volumes 2–10 µL, gradient elution over 9 min.
• MS system: Agilent Ultivo Triple Quadrupole with VacShield, Cyclone Ion Guide, Vortex Collision Cell and Hyperbolic Quads, operated in dMRM mode, 500 ms cycle time, source gases optimized for sensitivity.
• Software: Agilent MassHunter for method setup, data acquisition and reporting.

Main Results and Discussion

Sensitivity and Quantitation Limits

• Most mycotoxins achieved limits of quantitation at 1/20 of their assigned MRL, demonstrating outstanding sensitivity.
• Detection at 1/10 MRL in corn produced clear, reproducible signals with signal-to-noise ratios above 50.

Linearity and Precision

• Calibration curves for all compounds in each matrix exhibited R² values ≥ 0.99 over relevant concentration ranges.
• Precision at LOQ levels showed relative standard deviations below 10 %, indicating reliable quantification.

Matrix Effects and Robustness

• Optimized extraction and cleanup effectively minimized matrix interferences in complex samples like peanut and black pepper.
• The compact Ultivo design and advanced ion optics maintained performance without frequent maintenance interruptions.

Benefits and Practical Applications of the Method

The combined approach offers laboratories a high-throughput, space-saving solution for routine mycotoxin monitoring:

• Rapid 9-minute analysis cycle accelerates sample turnaround.
• Small instrument footprint and simplified maintenance suit nonexpert users.
• Wide dynamic range enables simultaneous quantification of multiple mycotoxins across diverse food types.

Future Trends and Opportunities

Emerging directions in mycotoxin analysis may include:

• Expansion to additional toxic metabolites and emerging fungal biomarkers.
• Integration with automated sample preparation for fully walk-away workflows.
• Application of high-resolution MS for untargeted screening alongside targeted LC/MS/MS quantification.
• Digital data management and AI-driven interpretation to streamline regulatory reporting.

Conclusion

The Agilent Ultivo Triple Quadrupole LC/MS system, paired with streamlined sample preparation and Agilent MassHunter software, delivers sensitive, precise and rapid quantification of regulated mycotoxins in complex food matrices. Its compact design, robust performance and user-friendly operation make it a powerful tool for routine food safety laboratories aiming to meet stringent regulatory requirements.

References

1. Bennett J.W.; Klich M. Mycotoxins. Clinical Microbiology Reviews 2003, 16(3), 497–516.
2. Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. Official Journal of the European Union L 364/5-24.
3. Commission Regulation (EU) No 105/2010 of 5 February 2010 amending Regulation (EC) No 1881/2006 as regards ochratoxin A. Official Journal of the European Union L 35/7-8.