Rapid Screening of 25 Environmental Hormones in Aquatic Products Using QuEChERS-Combined With Dispersive Liquid–Liquid Microextraction Based on Solidification of Floating Organic Droplet Coupled With UHPLC-Q/Orbitrap HRMS

Abstract

An innovative analytical method integrating QuEChERS with dispersive liquid–liquid microextraction using solidified floating organic droplets (DLLME-SFO) and ultra-high-performance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry (UHPLC-Q/Orbitrap HRMS) was developed for the rapid screening of 25 environmental hormones (EHs) in aquatic products. The method systematically optimized key parameters, including extraction solvent selection (1-undecanol), water volume (5 mL), pH (native pH 6.5), adsorbent combination (C18/PSA, 20 mg each), and salt addition (0% NaCl), achieving high sensitivity with detection limits of 0.1–0.4 µg/kg and recoveries of 68.2%–112.8%. The method demonstrated excellent linearity (R² > 0.995) across a concentration range of 0.4–40 µg/kg, with precision (intra- and inter-day RSDs < 15%) and manageable matrix effects (−19.5% to 10.9%).  The environmental impact assessment, conducted using the Analytical GREEnness Metric for Sample Preparation (AGREEprep score: 0.59) and Blue Applicability Grade Index (BAGI score: 80), confirmed that the proposed method satisfies both sustainability and practicality criteria for EHs analysis. Enhanced matrix purification and dual-polarity HRMS detection enabled accurate identification and quantification of structurally diverse EHs, including pesticides and pharmaceuticals. Validated in 60 commercial samples, the method detected prometryn residues in 11.7% of cases, demonstrating its practicality for monitoring EH contamination in complex aquatic matrices. This approach represents a significant advancement in multi-residue analysis, combining efficiency, sensitivity, and environmental compatibility.