Supramolecular solvent-based extraction of urinary biomarkers: a sustainable alternative for target analysis, suspect and non-target screening

Human exposure to hazardous chemical mixtures remains a critical public health challenge, but traditional biomonitoring approaches fail to capture this complexity. Suspect and non-target screening (SNTS) via high-resolution mass spectrometry (HRMS) can address this gap, but sample preparation remains a bottleneck. This study introduces supramolecular solvents (SUPRAS) as a sustainable, alternative extraction method for exposome research. A SUPRAS-based protocol (which employed 400 μL of tetrahydrofuran as dispersive solvent and 100 μL of 1-hexanol as amphiphile agent) was validated for the analysis of 11 representative compounds belonging to three different classes–bisphenols, phthalates, and per- and polyfluoroalkyl substances (PFAS)–in human urine. Compared to a conventional solid-phase extraction (SPE) clean-up method, the SUPRAS approach achieved higher average extraction recoveries for all target analytes (75 % for SUPRAS vs. 65 % for SPE), great precision (RSD < 25 %) and sensitivity (LOQs 0.04–1 ng mL−1). SUPRAS protocol offered significant advantages, including the reduction of organic solvent consumption and broad-spectrum analyte coverage in a single step, making it particularly suitable for SNTS applications. The non-selective nature of SUPRAS extraction method enabled the identification of 27 additional compounds through HRMS, compared to SPE's 11 unique detections. Chemical space evaluation demonstrated that SUPRAS extraction covered a wider LogP range (1.6–9.6) than SPE (1.6–7.4), highlighting its efficiency in detecting more hydrophobic substances than conventional SPE clean-up approaches. This work establishes SUPRAS as a transformative tool for exposome research, combining sustainability with high efficiency. By improving detection of complex mixtures and emerging chemical risks, the method advances public health biomonitoring. Its adaptability to high-throughput workflows positions SUPRAS as a critical innovation for large-scale exposure assessment.