High Throughput Quantitative Analysis of Multi-mycotoxin in Beer-based Drinks using UHPLC-MS/MS

Importance of the Topic

Mycotoxins are toxic secondary metabolites produced by fungi and can contaminate cereal grains used in beer production. Even at low levels, these compounds pose health risks, making their rapid and reliable quantification essential for ensuring consumer safety and regulatory compliance in the beverage industry.

Objectives and Study Overview

This study aimed to develop a high-throughput UHPLC-MS/MS method to simultaneously quantify 14 major mycotoxins in beer‐based beverages. Specific goals included achieving high sensitivity, minimizing autosampler carryover, and reducing total analysis time to support routine quality control.

Methodology and Instrumentation

Sample Preparation:

• Solid-phase extraction (SPE) cleanup of beer samples spiked with mycotoxin standards.

Chromatography:

• UFLC system with YMC-Triart C18 column (100 × 2.0 mm, 1.9 µm).
• Mobile phase A: 10 mM ammonium acetate in water; B: 2% acetic acid in methanol.
• Gradient from 2% to 85% B over 8 min, total run time 11 min; flow rate 0.4 mL/min; column at 40 °C.

Mass Spectrometry:

• Shimadzu LCMS-8030 triple quadrupole with ESI source.
• Positive/negative fast polarity switching (15 ms) in MRM mode.
• Individual MRM transitions optimized for 14 mycotoxins (e.g., AFB1 313.00>241.05, DON 355.10>295.15).

Carryover Mitigation:

• Shimadzu SIL-30AC autosampler with inner and outer needle rinse.
• Optimized rinse solvent: 1% formic acid aqueous/methanol/acetonitrile/isopropanol (1:1:1:1).

Main Results and Discussion

Separation and Sensitivity:

• All 14 mycotoxins separated within 11 minutes with sharp peaks for fumonisins and trichothecenes.
• Calibration linearity r2 = 0.994–0.999 over relevant concentration ranges (0.4–100 ppb).

Carryover Performance:

• Blank injections after 100 ppb standards showed negligible fumonisin carryover, confirming the effectiveness of the dual-rinse protocol.

Application to Commercial Samples:

• Analysis of 14 beer‐based drinks from 11 countries found trace levels (<5 ppb) of DON, HT-2, OTA, FMB1 and FMB2; other toxins were below detection.
• Overall contamination levels indicate a low health risk to consumers.

Benefits and Practical Applications

This rapid UHPLC-MS/MS workflow enables beverage manufacturers and testing laboratories to:

• Perform high‐throughput screening for multiple mycotoxins in a single run.
• Maintain stringent sensitivity and reproducibility for QA/QC.
• Reduce downtime and minimize sample carryover risks.

Future Trends and Potential Applications

Anticipated developments include:

• Expansion to other food and beverage matrices (e.g., juices, cereals).
• Integration with automated sample preparation and data processing using AI algorithms.
• Enhanced sensitivity via high-resolution mass analyzers or novel ionization techniques.
• Development of multiplexed methods for even broader contaminant panels.

Conclusion

A robust UHPLC-MS/MS method for simultaneous quantification of 14 mycotoxins in beer‐based drinks was established. The protocol delivers fast analysis, excellent linearity, and negligible carryover, supporting routine monitoring and ensuring consumer safety.

Reference

Masayoshi Tamura, Keiko Matsumoto, Jun Watanabe, Junko Iida, Yasushi Nagatomi, Naoki Mochizuki. High Throughput Quantitative Analysis of Multi‐mycotoxin in Beer‐based Drinks using UHPLC‐MS/MS. IMSC 2012 PTh-177, Shimadzu Corporation.