Identification of hazardous compounds in food matrices through integrated biological and chemical approaches focused on High-Performance Thin-Layer Chromatography

Food and food-related products may contain hazardous compounds from raw materials, processing, or packaging, introducing safety uncertainties in the food chain. Identifying toxicologically relevant chemicals (e.g., genotoxic/endocrine disruptors) requires efficient screening using integrated approaches [ 1 ]. The identification and characterization of potential hazards in extracts using traditional bioassays exhibits some limitations [ 2 ]. Therefore, approaches including fit-for-purpose bioassays for an efficient screening have been promoted. For this, a procedure combining high-performance thinlayer chromatography (HPTLC) bioautography (p-YES/Umu-C assay) was developed [ 3 ] [ 4 ]. Thus, a method combining high-performance thin-layer chromatography (HPTLC) bioautography (p-YES/Umu-C assay) with high-resolution mass spectrometry (HRMS) and microfractionation enables targeted isolation of bioactive compounds. Traditional TLC-MS desorption is a valuable tool however, is limited by resolution and metabolite yield, whereas HRMS-based metabolite profiling of crude extracts—paired with HPTLC bioactivity-guided fractionation—preserves sensitive bioactivity while improving chromatographic resolution. This workflow allows MS dereplication of known compounds and upscaling to HPLC semi-prep with dry load injection for targeted isolation in a single step [ 5 ]. Building on the success of the conclusive application to soy-based ingredients, the methodology was further extended to encompass other food-related complex matrices. Finally, the effectiveness of integrated approaches in contributing to the identification of potential hazardous compounds is promising. Publication History Article published online: 05 March 2026 © 2026. Thieme. All rights reserved. Georg Thieme Verlag KG Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany