Comprehensive pollutant screening in honey using integrated targeted and non-targeted LC-Q-Orbitrap HRMS

This study presents a comprehensive analytical framework combining targeted and non-targeted screening for pollutant residues in honey using liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS). A validated method targeting 30 veterinary drugs was extended to screen 300 honey samples. The analytical workflow employed Compound Discoverer software for data processing, integrating multiple database searches with stringent identification criteria including accurate mass (< 5 ppm), retention time (± 0.1 min), and MS2 fragmentation patterns. The screening identified 58 distinct compounds across diverse chemical classes. Pollutant residues included caffeine (69.7%), dulcin (61.7%), phenazone (41.0%), and multiple antibiotics, with sulfamethoxazole present in 20.0% of samples. Pesticide contamination was moderate, including acetamiprid (9.0%) and the neonicotinoid metabolite desnitro-imidacloprid (2.7%). Additionally, the non-targeted workflow captured 26 endogenous authenticity markers spanning flavonoids, phenolic acids, amino acids, organic acids, and sugar markers. Notably, aflatoxin B1 was detected in 2.7% of samples, alongside various mycotoxins and natural products. The wide concentration ranges observed, spanning 3-4 orders of magnitude, suggest both point-source and background environmental contamination. This integrated targeted/non-targeted approach demonstrates honey's utility as a biomonitoring matrix while revealing extensive contamination that warrants enhanced regulatory oversight.