Comparison of Analytical Results of Reference / QC Material Cleaned-up by CrossTOX® Column

Importance of the Topic

Mycotoxins are toxic secondary metabolites produced by fungi that frequently contaminate cereals, nuts, fruits, spices and animal feed. Their co-occurrence in diverse matrices poses a significant threat to food safety, animal health and agricultural economics. Efficient removal of matrix interferences prior to analysis improves precision, reduces solvent consumption and prolongs instrument uptime in high-throughput laboratories.

Objectives and Study Overview

This study evaluates the performance of the CrossTOX® cleanup column for multi-toxin sample preparation in comparison to established immunoaffinity (IAC) and solid-phase extraction (SPE) methods. Certified reference materials and quality control samples covering aflatoxins, ochratoxin A, deoxynivalenol (DON), zearalenone (ZEA), fumonisins, T2/HT2 toxins and citrinin were processed across cereals, nuts, fruits, spices and feed matrices. Analytical outputs were assessed by HPLC-FLD, HPLC-UV and LC-MS/MS for recovery, accuracy and matrix interference.

Methodology

Samples were extracted with optimized solvent mixtures—acetonitrile/water/acetic acid (84/15/1, v/v/v) for CrossTOX® and matrix-specific solvents for immunoaffinity and SPE. Crude extracts were filtered and directly loaded onto the CrossTOX® column at 1 mL/min. Parallel cleanup with IAC (AflaCLEAN, OtaCLEAN, Afla-OtaCLEAN) and SPE (ZeaCLEAN SMART, DONeX) followed manufacturer protocols. Eluates were collected, evaporated when required and reconstituted for LC-MS/MS or HPLC-FLD/UV analysis.

Used Instrumentation

• UHPLC-MS/MS (Thermo Fisher Scientific)
• HPLC-FLD
• HPLC-UV

Key Results and Discussion

Across all matrices, CrossTOX® yielded recoveries within interlaboratory acceptance ranges. In pepper and spices, aflatoxins B1, B2, G1 and G2 recovered between 80–100% of assigned values. Ochratoxin A in cookie flour matched IAC recoveries at 90–95%. In cereal-based feed, DON, ZEA and AFB1 results aligned closely with DONeX and ZeaCLEAN SMART, with deviations under 15%. CrossTOX® consistently reduced co-eluting matrix peaks, enhancing signal-to-noise ratios in LC-MS/MS.

Benefits and Practical Applications of the Method

CrossTOX® offers a universal, non-dispersive cleanup suitable for a broad spectrum of mycotoxins and sample types. The single-step protocol reduces solvent usage, eliminates extensive defatting or dilution steps and decreases reliance on multiple specific immunoaffinity columns. Laboratories can minimize internal standard requirements and instrument maintenance downtime, supporting high-throughput screening and regulatory compliance.

Future Trends and Opportunities

Advances may include automated CrossTOX® formats compatible with robotic platforms, expansion of sorbent chemistries to emerging mycotoxins, integration with high-resolution mass spectrometry for untargeted screening and coupling with data-driven algorithms for real-time quality control. Miniaturized column formats could further reduce solvent consumption and enhance green analytical chemistry goals.

Conclusion

The CrossTOX® cleanup column delivers robust, reproducible and accurate multi-mycotoxin cleanup across diverse matrices. Its performance matches or exceeds conventional IAC and SPE methods, while simplifying workflow, reducing costs and boosting analytical throughput.