Validation of a Low-Cost and Highly-Sensitive Method for Determination of Eighteen Mycotoxins in Food Matrices Using SPE and LC/MS/MS

Significance of the Topic

This work addresses the critical need to monitor mycotoxins in food products, as these fungal metabolites pose mutagenic and carcinogenic risks to human health. Regulatory bodies such as the European Union enforce strict maximum levels for multiple mycotoxins, creating demand for cost-effective, sensitive and multi-analyte analytical methods.

Objectives and Study Overview

The study aimed to develop and validate a solid-phase extraction (SPE) approach coupled with triple-quadrupole LC-MS/MS for simultaneous determination of 18 EU-regulated mycotoxins in diverse food matrices. Low limits of quantitation (LOQs), high recoveries and robust repeatability were targeted across five representative agricultural products.

Instrumentation Used

• Shimadzu LCMS-8060 triple-quadrupole LC-MS/MS system
• Shimadzu GLC MastroTM PFP column (100 mm × 2.1 mm, 3 µm)
• ISOLUTE® Myco SPE cartridges (Biotage)

Methodology and Sample Preparation

Samples (1 g) of rice, barley, wheat flour, cashew and corn were extracted by acidified water or water/ACN, centrifuged, diluted with water and loaded onto SPE cartridges. Two SPE protocols were optimized to recover polar and less-polar mycotoxins separately. Eluates were dried under nitrogen, reconstituted in 10% ACN and injected (10 µL) onto the PFP column under gradient elution with ammonium fluoride buffers. Electrospray ionization in positive/negative mode with MRM transitions was automated via LabSolutions software.

Main Results and Discussion

Calibration curves (0.01–500 ng/mL) exhibited r2 > 0.998 for all compounds. LOQs ranged from 0.01 to 1.0 ng/mL, and LODs were typically one-third of LOQs. Repeatability (%RSD, n=6) was below 12.3%. Recovery in spiked matrices ranged 65.9–143.0% except for lower recoveries of NIV in wheat and inflated OA recoveries. Analysis of ten commercial samples detected fumonisins and aflatoxins in two samples with concentrations up to 178 ng/g.

Advantages and Practical Applications

• Cost-effective SPE cartridges replace expensive immunoaffinity columns
• Simultaneous analysis of 18 mycotoxins saves time and solvent consumption
• High sensitivity meets EU regulatory limits across diverse food matrices

Future Trends and Applications

Further enhancements may include automation of SPE workflows, expansion of analyte panels to emerging toxins, integration with high-resolution MS for non-target screening, and miniature “lab-on-a-chip” devices for on-site testing in supply chains.

Conclusion

The validated SPE–LC-MS/MS method demonstrates excellent linearity, sensitivity and accuracy for 18 regulated mycotoxins in multiple food matrices. Its low cost and robust performance make it suitable for routine quality control and compliance monitoring.

References

1. Commission Regulation (EC) No 1881/2006 on setting maximum levels for certain contaminants in foodstuffs. Official Journal of the European Union, L 364/5.
2. Baker D., Titman C., Loftus N., Horner J. Multi-residue analysis of 18 regulated mycotoxins by LC-MS/MS. ASMS 2017, Poster Session TP 185.