Multi-mycotoxin determination in human urine by UHPLC-MS/MS: An environmentally friendly and high-throughput approach

Mycotoxins, naturally occurring secondary metabolites produced by fungi, are globally prevalent in food and pose serious health risks to humans due to their toxicity, which include carcinogenic, mutagenic, and neurotoxic effects. Due to their diverse biochemical nature, these compounds follow distinct metabolic pathways in the human organism, with urinary excretion being their principal elimination route. Harmonization in human biomonitoring (HBM) is essential for comparable data across studies and regions, while trustworthy and validated analytical methodologies are vital for meaningful health insights. There's also a growing need for innovative, eco-friendly analytical techniques. In this context, we have developed and validated an eco-friendly analytical procedure for the rapid and simultaneous detection of multiple mycotoxins in urine. This approach utilizes a straightforward "dilute-and-shoot" sample preparation, combined with ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The method targets twelve mycotoxins, achieving relative recoveries between 79.2 % and 113.5 %, with precision values consistently below 20 % for all analytes. The quantification range includes concentrations typically found in urine, with limits of quantification (LoQs) ranging from 0.005 to 0.5 ng/mL. Our evaluation demonstrates that this methodology offers a more sustainable and practical alternative to mycotoxin quantification in urine, with AGREEprep and BAGI scores of 0.73 and 85.0, respectively, and can be widely implemented in HBM programs. Furthermore, its application to 25 urine samples from adults residing in the Valencian Community (Spain) confirmed its effectiveness for HBM studies. Finally, a non-negligible level of exposure to these compounds within the population studied was observed.