Enhanced determination of 85 mycotoxins in challenging root and rhizome herbal medicines using online segmented multi-dimensional liquid chromatography-tandem mass spectrometry.

Sensitive and accurate detection of mycotoxin contaminations is crucial for ensuring safety of herbal medicines. However, their inherent chemical complexity poses significant challenges for reliable method development. Herein, an innovative segmented multi-dimensional liquid chromatography-tandem mass spectrometry (sMD-LC-MS/MS) method was developed for detection of 85 mycotoxins in complex root and rhizome herbs. By integrating multiple heart-cutting, segmented trapping, and dual-mode second-dimension separations, it resolved critical co-elution issues from conventional 1D analysis. Coupled with a simple "dilute-and-shoot" workflow, the system eliminated offline purification, enhancing throughput. Method validation across six representative herbal matrices demonstrated that this "panoramic" strategy significantly outperformed conventional LC-MS/MS method and achieved improvements in qualitative reliability, quantitative accuracy, and detection sensitivity. It provided a systematic solution for high-throughput, highly reliable detection of trace-level, multiclass mycotoxins in challenging herbal matrices.