Non-Targeted LC-Q-Orbitrap HRMS Screening of Veterinary Drugs, Pesticides, and Emerging Contaminants in Animal-Derived Foods and Honey

Background Veterinary drug residues, pesticides, and chemical contaminants in animal-derived foods pose public health risks, including antimicrobial resistance development and consumer exposure to banned or withdrawn substances. Conventional targeted methods are inherently limited to predefined compounds and may overlook emerging or unexpected contaminants. Many developing countries still lack comprehensive multi-residue surveillance across diverse food matrices, highlighting the need for analytical frameworks adaptable to regulatory laboratories. Objective To evaluate an LC-Q-Orbitrap HRMS framework combining reference-standard-assisted confirmation with non-targeted screening for the detection and identification of veterinary drug residues, pesticides, and chemical contaminants across multiple animal-derived food matrices. Methods A streamlined extraction protocol coupled with liquid chromatography–quadrupole-Orbitrap high-resolution mass spectrometry was applied to 450 retail samples collected in Egypt, covering nine matrices (bovine liver, kidney, meat, heart, raw bovine milk, chicken breast, chicken eggs, honey, and Nile tilapia (Oreochromis niloticus)). Confirmation used 36 certified reference standards; non-targeted screening used Compound Discoverer™ with spectral database matching and in silico fragmentation prediction. Results Nine compounds were confirmed at Level 1 (confirmed structure via reference standard match) according to the Schymanski identification confidence framework, accounting for 74 detection events, most frequently oxytetracycline (18 detections) and trimethoprim (17 detections). Non-targeted screening revealed 45 additional compounds identified at Level 2a (probable structure based on library spectrum match), including antimicrobials, β-agonists, pesticides, and pharmaceuticals. Phenacetin and tapentadol were each detected in 48 samples, and fenobucarb in 30% of milk samples. Liver and honey showed the greatest diversity of detected compounds. Conclusion HRMS-based non-targeted screening extends analytical coverage beyond predefined reference-standard panels in multi-matrix food surveillance. Highlights LC-Q-Orbitrap HRMS detected 54 compounds across nine animal-derived food matrices—nine Level 1 confirmations and 45 Level 2a probable structures—extending chemical surveillance beyond reference-standard panels.