The Benefits of ACQUITY Premier UPLC for Multi-Mycotoxin Methods

Significance of the Topic

Multi-mycotoxin analysis is essential for ensuring food and feed safety, given the diversity and toxicity of fungal metabolites that can co-occur in agricultural products. Robust, high-throughput methods are needed to address challenges such as uneven detector response, chromatographic carryover, and matrix-induced ion suppression, which can compromise accuracy and reliability in routine testing.

Objectives and Study Overview

This study aimed to evaluate the performance of Waters’ new ACQUITY Premier UPLC System—featuring MaxPeak High Performance Surfaces (HPS) technology—when transferring an established dilute-and-shoot LC-MS/MS multi-mycotoxin method. Key goals included comparing carryover levels, sensitivity, linearity, precision, peak shape, and retention time stability against a conventional ACQUITY UPLC I-Class PLUS system under typical analytical sequences for cereal samples.

Methodology

Sample preparation followed a dilute-and-shoot protocol for wheat, oat, and wholemeal flour: 5 g of sample was extracted with acetonitrile/water/acetic acid (79:20:1 v/v/v), vortexed, centrifuged, filtered, and diluted to achieve a 1:20 overall dilution. An internal standard mixture of ^13C-labelled mycotoxins was added prior to analysis. Calibration curves were prepared in water/acetonitrile (95:5), spiked with internal standards.

Applied Instrumentation

• Liquid Chromatography: Waters ACQUITY Premier UPLC and ACQUITY UPLC I-Class PLUS, Flow Through Needle injector with 15 μL needle and 50 μL extension loop
• Columns: ACQUITY Premier BEH C18 1.7 μm and standard BEH C18 1.7 μm (40 °C)
• Mobile Phase: A – 1 mM ammonium acetate in water with 0.3% acetic acid and 0.1% formic acid; B – methanol with 0.3% acetic acid and 0.1% formic acid
• Mass Spectrometer: Xevo TQ-S micro (ESI±, MRM mode)
• Other Conditions: Flow rate 0.40 mL/min; injection volume 5 μL; sample temperature 15 °C; desolvation 600 °C, gas flows 50 L/hr cone and 1100 L/hr desolvation

Main Results and Discussion

Carryover experiments using fumonisin B2 (2000 µg/kg standard) showed that the Premier system reduced carryover by ~80% compared to the I-Class. On ACQUITY Premier, FB2 levels in the first blank were at or below the lower limit of quantification (LLOQ = 15 µg/kg), reaching non-detectable by the second blank. Performance metrics for 12 regulated mycotoxins met SANTE/12089/2016 guidelines:

• Linearity: R² > 0.99, residuals within ±20%
• Precision: RSD <10% for spiked and incurred samples
• Retention Time Reproducibility: deviations <0.006 min
• Peak Shape: symmetric Gaussian profiles without tailing

Minor improvements in signal-to-noise ratios were observed on the Premier system for certain analytes at their LLOQ.

Benefits and Practical Applications

• Significant reduction in analyte carryover, eliminating the need for metal chelators in mobile and wash solvents
• Improved analytical throughput by reducing the number and duration of wash cycles
• Maintained or enhanced method sensitivity, precision, and reproducibility
• Direct transfer of existing multi-mycotoxin workflows with minimal method adjustment

Future Trends and Possibilities

Advances in low-adsorption materials and high-performance surface chemistries will continue to mitigate interaction-driven carryover. Integration of UPLC systems with real-time QC monitoring and automated data correction could further streamline multi-residue analysis. Expanding high-throughput platforms to include emerging mycotoxin markers and adapting to alternative food matrices remain promising areas for development.

Conclusion

The ACQUITY Premier UPLC System, enabled by MaxPeak HPS technology, delivers equivalent or superior multi-mycotoxin method performance compared to conventional UPLC platforms, with a pronounced decrease in carryover, robust quantitation, and higher sample throughput, fulfilling stringent regulatory requirements for routine food safety testing.

References

1. Dreolin N., Stead S. LC-MS/MS Method Development and Validation for Regulated Mycotoxins in Cereal Flours Using Simplified Sample Preparation on Xevo TQ-XS. Waters Application Note, 2019, 720006685.
2. Dreolin N. et al. Development of a Multi-Toxin UPLC-MS/MS Method for 50 Mycotoxins and Tropane Alkaloids in Cereals. Waters Application Note, 2021, 720007476.
3. Dreolin N. et al. Improve the Robustness of an LC-MS/MS Method for Multiple Mycotoxins in Food Matrices. Waters White Paper, 2022, 720007521EN.
4. Dreolin N. Carryover in UPLC Methods: Case Study of Fumonisins B2. Waters White Paper, 2020, 720006826EN.
5. Walter T. H. et al. Low Adsorption UPLC System and Columns Based on MaxPeak HPS: ACQUITY Premier Solution. Waters White Paper, 2021, 720007128EN.
6. SANTE/12089/2016 Guidance Document on Identification of Mycotoxins in Food and Feed. Implemented 01/01/2017.
7. Tamura M. et al. Minimization of Carryover for High-Throughput LC-MS/MS of 14 Mycotoxins in Corn Grits. Journal of Separation Science, 2014, DOI:10.1002/jssc.201400099.