Extraction of Multiple Mycotoxins From Cereal Based Infant Food Using ISOLUTE® Myco prior to UHPLC MS/MS Analysis

Importance of the Topic

Mycotoxin contamination in cereal‐based infant foods poses significant health risks due to the toxicity of compounds such as aflatoxins, ochratoxin A, fumonisins and trichothecenes. Regulatory bodies enforce strict maximum residue limits to protect vulnerable populations. A robust sample preparation and sensitive analytical protocol are essential for reliable monitoring of multiple mycotoxin classes in complex food matrices.

Objectives and Study Overview

This study aimed to develop and validate a streamlined solid phase extraction (SPE) workflow using ISOLUTE® Myco columns for simultaneous cleanup of diverse mycotoxins in infant cereal. The extracts were analyzed by UHPLC‐MS/MS, targeting nine major mycotoxins. Key performance metrics—recovery, repeatability, limits of quantitation (LOQs) and linearity—were assessed against European regulatory criteria.

Methodology and Instrumentation

Sample preparation comprised:

• Grinding 5 g of infant cereal with salt addition
• Extraction in 60 % aqueous methanol, shaking and centrifugation
• Dilution of supernatant with water, further centrifugation

SPE cleanup on ISOLUTE Myco 60 mg/3 mL columns:

• Conditioning with acetonitrile, equilibration with water
• Loading diluted extract (2 × 2.5 mL)
• Sequential washes: water, then 10 % acetonitrile
• Drying under vacuum, elution in acidified organic solvents
• Evaporation and reconstitution in acetic acid–methanol–acetonitrile–water mixture, final filtration

Used Instrumentation

• UHPLC: Shimadzu Nexera with Kinetex XB-C18 column (50 × 2.1 mm, 1.7 μm)
• MS/MS: AB Sciex Triple Quad 5500 with Turbo-V ESI source, operated in dual‐polarity MRM mode

Main Results and Discussion

The method achieved:

• LOQs of 0.2 µg kg⁻¹ for aflatoxins and ochratoxin A, 2 µg kg⁻¹ for T-2/HT-2, 2 µg kg⁻¹ for zearalenone and 2 µg kg⁻¹ for fumonisin B1
• Recoveries between 65 % and 98 % across all analytes, meeting EU MRL criteria
• Excellent repeatability (%RSD <11 %) and linearity (r² >0.989)

Extracted ion chromatograms demonstrated clear separation within an 8.2 min run time with retention times from 2.8 to 5.4 min.

Benefits and Practical Applications

The single‐product SPE approach simplifies multi‐mycotoxin analysis, enhances throughput and reduces solvent consumption. High sensitivity and reproducibility make it suitable for quality control labs, regulatory testing and research on infant food safety.

Future Trends and Opportunities

Advances may include expansion to other complex matrices, incorporation of high‐resolution mass spectrometry for untargeted screening, automation of SPE workflows and development of greener extraction solvents. Integration with digital data analysis platforms will further streamline compliance monitoring.

Conclusion

The ISOLUTE Myco SPE combined with UHPLC‐MS/MS provides a reliable, high‐throughput solution for simultaneous quantitation of multiple mycotoxins in cereal‐based infant food, fulfilling regulatory requirements and supporting food safety initiatives.