Mycotoxin Workshop 2018: Fully Automated Mycotoxin Analysis From Extract to Chromatogram

Importance of the topic

The accurate detection of mycotoxins in food and feed is critical for consumer safety and regulatory compliance. Manual sample cleanup methods are time-consuming and prone to variability, especially when ultra-trace sensitivity is required for products like baby food.

Study objectives and overview

This work demonstrates a fully automated workflow that integrates sample extraction, immunoaffinity cleanup, and chromatographic analysis to achieve ppt and ppq-level sensitivity. The methodology employs the FREESTYLE ThermELUTE™ robotic system in combination with SMART columns and HPLC-FLD detection.

Methodology and instrumentation

The automated process consists of:

• Extraction of samples with methanol/water (80/20, v/v) and optional n-hexane defatting for fatty matrices;
• Automated loading of 0.08–10 g matrix equivalents onto SMART immunoaffinity columns via the FREESTYLE ThermELUTE™ platform at up to 3 mL/min;
• Washing to remove matrix interferents followed by thermal denaturation elution of bound mycotoxins;
• Direct quantitative injection into an HPLC system equipped with fluorescence detection HPLC-FLD;
• Parallel processing capability, where the next sample is prepared while the current one is being analyzed.

Main results and discussion

Key performance metrics include:

• Limits of detection ranging from 0.7 ppt for Aflatoxin M1 to 10 ppt for Ochratoxin A, with LOQs up to 31 ppt;
• Highly linear calibration curves across a broad concentration range and various matrices;
• Recovery rates exceeding 90% for multiple toxins in matrices such as pistachio, curcuma, corn, black tea, raisins, milk products, coffee, and chili;
• Reproducible chromatographic profiles for low-level spiked samples, demonstrating robustness and high throughput.

Benefits and practical applications

This automated approach offers:

• Significant reduction of manual preparation steps, lowering labor and potential for error;
• Increased sample throughput and lower per-sample costs through parallel processing;
• Applicability to diverse food and feed matrices from baby food to animal feed, ensuring compliance with stringent safety standards;
• Trace-level sensitivity that meets or exceeds regulatory limits.

Future trends and potential applications

Emerging directions include:

• Integration with LC-MS/MS for expanded analyte coverage and confirmatory testing;
• Development of multiplexed immunoaffinity formats for simultaneous detection of multiple mycotoxins;
• Implementation of real-time data analytics and machine learning to optimize workflows and predict quality outcomes;
• Wider adoption in high-throughput laboratory environments and regulatory monitoring programs.

Conclusion

The FREESTYLE ThermELUTE™ platform, combined with SMART immunoaffinity columns and HPLC-FLD, delivers a fully automated, sensitive, and robust solution for mycotoxin analysis, reducing labor and cost while ensuring compliance with stringent safety requirements.

Reference

Wuppermann FN et al., Fully Automated Mycotoxin Analysis from Extract to Chromatogram, LCTech GmbH.