Mycotoxins are among the most closely watched contaminants in the food and feed industry, and for good reason, since they are considered carcinogenic, hepatotoxic, and immunosuppressive. The Food and Agriculture Organization (FAO) estimates that more than a quarter of the world’s food crops are affected by these fungal metabolites, while industry surveys consistently find that over 90% of contaminated samples contain not one but several mycotoxins at once. In practice, roughly one in four raw materials entering the supply chain fails to meet regulatory requirements. For any laboratory responsible for food safety, this is not an abstract statistic, it is a daily challenge.

Meanwhile, the pressure is on. Warmer, wetter and more unpredictable weather is expanding the range and intensity of toxin-producing fungi such as Aspergillus and Penicillium. Recent European research points to aflatoxin contamination of crops (like maize) moving steadily northwards as temperatures climb, turning what was once a largely Southern-European concern into a continent-wide one. At the same time, regulators are tightening their grip: the maximum levels set out in Commission Regulation (EU) 2023/915 and its 2024/1756 amendment are demanding, especially for sensitive categories such as cereal-based baby food, where compliance is measured in fractions of a microgram per kilogram.

The real bottleneck: not detection, but sample preparation

Most laboratories already have capable LC and LC-MS instrumentation. The constraint is everything that happens before the sample reaches the column. Immuno-affinity chromatography (IAC) is the established, selective way to clean up and concentrate mycotoxins from complex extracts.

However, the classic workflow is slow and labor-intensive: large load volumes, methanol elution step, followed evaporation and reconstitution before a single injection can be made. Every manual step adds time, consumes solvent and introduces variability in flow rate and volume handling. All of which are significantly impacting recovery and repeatability.

Multiply that by the sheer number and type of samples a lab typically handles (across grains, nuts, coffee, cocoa, spices, beverages and infant cereals) and sample preparation becomes the rate-limiting step for the entire operation.

Two applications, one streamlined workflow

We have built and validated an automation-ready mycotoxin sample-preparation workflow around two of the most requested analyses in the field:

• Ochratoxin A (OTA): validated across red wine, lager, peanuts, Brazil nuts, coffee, cocoa powder and spices using OtaCLEAN SMART cartridges.
• Aflatoxins B1, B2, G1, G2 (AB1, AB2, AG1, AG2): validated in corn, nuts, chocolate and olive oil using AflaCLEAN SMART cartridges, with reagent-free UVE photochemical derivatization to boost the fluorescence of the most toxic congeners.

What actually makes our approach different

Selective immuno-affinity cartridges are not new, and on their own they are not unique. They rely on well-established monoclonal-antibody chemistry. The difference lies in how that chemistry is packaged and run:

• Miniaturized SMART cartridges cut the load volume dramatically (around 80% less than classic 3 mL formats), thereby reducing sample and solvent consumption and shortening the most time-consuming step of the analysis.
• Automation on the Gerstel MultiPurposeSampler (MPS) or the LCTech FREESTYLE turns conditioning, loading, washing, and elution into a hands-off, walkaway process that runs reproducibly around the clock.
• Thermal elution (ThermELUTE / ThermELUTE Light) releases the toxins into a water-based, LC-compatible solvent instead of methanol. Because there is no evaporation or reconstitution, large extract volumes can be injected directly onto the LC, preserving sensitivity.

The final result is a typical IAC clean-up time of well under 15 minutes per sample, with controlled flow and volume handling that only automation can guarantee.

Sensitivity where it matters most

Direct, high-volume injection is what makes the workflow capable at baby-food compliance levels. On LC-FLD, our setup comfortably meets the strictest thresholds in the regulation: ochratoxin A at 0.5 µg/kg and aflatoxins at 0.1 µg/kg in cereal-based matrices. The same LC-ready extracts can be taken straight to LC-MS/MS when the application calls for the highest selectivity and confirmation (i.e. the recognized gold standard for mycotoxin quantification).

Oatmeal is a classic cereal-based baby food and therefore falls under the strictest EU limit for ochratoxin A, just 0.5 µg/kg. To show how the workflow performs at this level, we spiked an oatmeal sample at exactly that threshold and ran it alongside an unspiked blank. The OTA peak is clean and fully resolved, with the toxin detected at 0.07 µg/kg and a recovery of 95%. The same holds for the aflatoxins: in oatmeal spiked at the EU baby-food limit of 0.1 µg/kg, aflatoxins B1, B2, G1, G2 (AB1, AB2, AG1, AG2) are cleanly resolved, with aflatoxin B1 (AB2) detected at 0.10 µg/kg and a recovery of 94%.

Crucially, our workflow holds up in the matrices most laboratories dread. Across the full validation set (baby-food cereals, beverages, nuts, coffee, cocoa and high-fat samples such as olive oil and chocolate) recoveries stayed within a dependable 69-104% for both ochratoxin A and the aflatoxins. Spices are the toughest test: our work on chili flakes (85% recovery) and black pepper (72%) picked up the matrix-driven false positive that catches labs out and showed how proper clean-up resolves it. Coffee, cocoa and fatty matrices are all handled by the same setup, so a single method covers the entire spread of food and feed samples.

Conclusion

Mycotoxin analysis no longer has to be slow to be reliable. By miniaturizing the cartridge, automating every handling step and replacing methanol elution with direct-injection thermal elution, you gain throughput, robustness and walkaway time without trading away the sensitivity that baby-food compliance demands.

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