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

Significance of the Topic

Mycotoxins are toxic secondary metabolites frequently contaminating grains and grain-derived beverages. Rapid and sensitive quantification in complex matrices like beer is essential to ensure consumer safety and comply with quality standards.

Study Objectives and Overview

This study aimed to develop a high throughput UHPLC-MS/MS method for simultaneous quantification of fourteen mycotoxins in beer-based drinks. The objectives included optimizing chromatographic separation, maximizing sensitivity, and suppressing system carryover during automated analysis.

Methodology and Instrumentation

Sample Preparation involved solid phase extraction of beer samples to isolate and concentrate mycotoxins. Chromatographic separation employed a Triart C18 column (100 mm×2.0 mm, 1.9 µm) at 40°C with a binary gradient of 10 mM ammonium acetate in water (A) and 2% acetic acid in methanol (B) at 0.4 mL/min, achieving full separation in 11 minutes. Mass spectrometric detection used ESI in positive/negative MRM mode with 15 ms ultra fast polarity switching. Rinse protocols for the UHPLC autosampler were optimized using mixed solvent containing 1% formic acid, effectively eliminating fumonisin carryover.

Used Instrumentation

• Shimadzu UFLC HPLC system
• Triart C18 column (100×2.0 mm, 1.9 µm)
• Shimadzu LCMS-8030 triple quadrupole mass spectrometer with ESI source
• Nexera SIL-30AC autosampler with inner and outer needle rinse capability

Main Results and Discussion

The optimized method resolved all mycotoxins within 11 minutes with excellent peak shape after pH adjustment of mobile phase to mitigate fumonisin tailing. Calibration curves across six concentration levels showed linearity (r2 ≥0.994) over typical regulatory ranges. The novel rinse protocol reduced fumonisin carryover to below detection limits in blank injections. Analysis of 20 commercial beer samples revealed sporadic low-level contamination by deoxynivalenol, HT-2 toxin, ochratoxin A, and fumonisins, all below 5 ppb.

Benefits and Practical Applications of the Method

• High throughput and short analysis time facilitate routine monitoring.
• Sub-ppb sensitivity supports compliance with strict food safety regulations.
• Robust autosampler cleaning minimizes false positives from carryover.
• Applicable to quality control in breweries and food safety laboratories.

Future Trends and Possibilities

Integration with high-resolution mass spectrometry could expand analyte scope and structural elucidation. Automation of sample prep and data processing will further increase throughput. Method adaptation to other beverage and food matrices can support comprehensive mycotoxin surveillance.

Conclusion

A rapid, sensitive UHPLC-MS/MS method was established for quantifying fourteen mycotoxins in beer-based drinks. Optimized chromatographic conditions and autosampler rinse strategies achieved reliable separation, low carryover, and excellent linearity, demonstrating suitability for routine food safety analysis.

References

• Tamura M. et al. High Throughput Quantitative Analysis of Multi-mycotoxin in Beer-based Drinks using UHPLC-MS/MS. ASMS 2012 WP27-580.