Development of a Multi-Toxin UHPLC-MS/MS Method for 50 Mycotoxins and Tropane Alkaloids in Cereal Commodities

Significance of the Topic

Mycotoxin contamination of cereal commodities poses serious risks to food safety and public health. Analytical methods capable of detecting multiple toxins simultaneously support regulatory compliance, quality assurance, and risk assessment. High-throughput, sensitive assays are essential for monitoring both regulated and emerging mycotoxins as well as tropane alkaloids in complex food matrices.

Objectives and Study Overview

This study aimed to extend an existing application note by developing a single-run UHPLC-MS/MS protocol for quantitation of over fifty mycotoxins and plant toxins in cereal-based foods. The method leverages the Xevo TQ-XS Mass Spectrometer’s sensitivity and introduces waters_connect quantitation software to streamline data processing and result review.

Methodology and Instrumentation

Sample Preparation

• Mixed flour matrix (wheat, barley, rice, maize) homogenized and extracted (5 g sample with 20 mL MeCN:H₂O:acetic acid, 79:20:1).
• Vortex mixing for 10 minutes, centrifugation at >5000 g for 6 minutes.
• Dilution of 150 µL supernatant into 1350 µL water (1:10 dilution, overall dilution factor 40), followed by glass fiber filtration.

Chromatography and Detection

• UHPLC separation using established reversed-phase method; Moniliformin noted as an exception due to high polarity.
• MRM acquisition with two transitions per analyte, retention time stability within ±0.03 minutes.
• Xevo TQ-XS Mass Spectrometer coupled to waters_connect software for quantitation workflow.

Calibration

• Solvent-based and matrix-matched standards prepared by serial dilution of stock mix.
• Calibration range spanned three orders of magnitude, R² values >0.99, residuals <20%.

Main Results and Discussion

The instrument limits of detection (i-LOD) reached as low as 0.0003 ng/mL for key aflatoxins. Method limits of quantification (m-LOQ) were 0.1 µg/kg for aflatoxins and ≤1 µg/kg for several other toxins including ochratoxins and tropane alkaloids. Most remaining analytes exhibited m-LOQs ≤10 µg/kg. Chromatograms demonstrated clear separation and consistent peak shapes. Matrix-matched calibration curves for aflatoxin B1 and ochratoxin A showed linear response with residuals within acceptance criteria.

Benefits and Practical Applications

This multi-toxin method offers a rapid dilute-and-shoot workflow reducing sample preparation complexity. High sensitivity enables extensive dilution, minimizing matrix effects and instrument contamination. The simultaneous analysis of diverse regulated and emerging toxins supports comprehensive screening in food and feed laboratories.

Future Trends and Potential Applications

Advancements may include integration of hydrophilic interaction chromatography for highly polar toxins, expanded coverage of masked mycotoxins, and incorporation of 13C-labelled internal standards for improved quantitation accuracy. Automation of data review and reporting within cloud-enabled platforms will further enhance throughput and traceability.

Conclusion

The developed UHPLC-MS/MS method on the Xevo TQ-XS platform delivers robust, sensitive, and high-throughput quantitation of over fifty mycotoxins and tropane alkaloids in cereals. Its simplified sample preparation, excellent detection limits, and streamlined data processing make it well suited for routine food safety testing and regulatory compliance.

Reference

Nicola Dreolin, Henry Foddy, Simon Hird, Peter Hancock, Timothy Jenkins. Development of a Multi-Toxin UHPLC-MS/MS Method for 50 Mycotoxins and Tropane Alkaloids in Cereal Commodities. Waters Corporation, 2022.