As one of the most important conjugated mycotoxins, zearalenone-14-glucoside (Z14G) has received widespread attention from researchers. Although the metabolism of Z14G in animals has been extensively studied, the intracellular toxicity and metabolic process of Z14G are not fully elucidated. In this study, the cytotoxicity of Z14G to human ovarian granulosa cells (KGN) and the metabolism of Z14G in KGN cells were determined. Furthermore, the experiments of co-administration of β-glucosidase and pre-administered β-glucosidase inhibitor (Conduritol B epoxide, CBE) were used to clarify the mechanism of Z14G toxicity release. Finally, the human colon adenocarcinoma cell (Caco-2) metabolism model was used to verify the toxicity release mechanism of Z14G. The results showed that the IC₅₀ of Z14G for KGN cells was 420 μM, and the relative hydrolysis rate of Z14G on ZEN was 35% (25% extracellular and 10% intracellular in KGN cells). The results indicated that Z14G cannot enter cells, and Z14G is only hydrolyzed extracellularly to its prototype zearalenone (ZEN) by β-glucosidase which can exert toxic effects in cells. In conclusion, this study demonstrated the cytotoxicity of Z14G and clarified the toxicity release mechanism of Z14G. Different from previous findings, our results showed that Z14G cannot enter cells but exerts cytotoxicity through deglycosylation. This study promotes the formulation of a risk assessment and legislation limit for ZEN and its metabolites.

Mycotoxins commonly contaminate grains and traditional Chinese medicinal materials, posing serious health risks to humans and animals. To address this issue, a magnetic microporous organic network (MMON) was synthesized via an in situ growth method and Sonogashira–Hagihara coupling for the simultaneous adsorption of seven mycotoxins, followed by UPLC-MS/MS detection. The optimized MMON featured a high surface area, uniform micropores, and rapid magnetic separation within 5 s. Structural and compositional analyses confirmed its tailored architecture, while DFT calculations revealed a pore confinement effect, π–π stacking, and hydrophobic interactions as the primary adsorption mechanisms. A magnetic solid-phase extraction (MSPE) method using 8 mg of MMON achieved adsorption equilibrium within 10 s in 5 mL of a 4 mg/L mycotoxin standard solution. The material maintained over 95% efficiency across ten reuse cycles at a low cost. Under optimal conditions, an MSPE-UPLC-MS/MS method with a low detection limit (0.002–0.15 μg/L), wide linear range (0.01–100.0 μg/L), large enrichment factor (20.1–21.9), low adsorbent dosage, and short extraction time was developed. The determination of mycotoxins in complex grain-based foods and herbal products was also realized with recoveries of 81.32% to 116.10%. This work offers a rapid, cost-effective, and high-throughput approach for mycotoxin detection, supporting quality control in food and herbal product safety.