Honey residue and contaminant assessments are extremely important, as they deal with food and therapeutic safety issues of a widely consumed animal product. Pyrrolizidine alkaloids are secondary plant metabolites used by bees in honey production. The toxicity of these alkaloids has been discussed in the literature and health risks have been noted, due to their occurrence in both honey and in pyrrolizidine alkaloid-producing plants. Quinolones comprise a class of antimicrobials associated with hypersensitivity reactions and bacterial resistance, one of the world's most significant public health problems. In this context, the aim of this study was to evaluate the simultaneous presence of pyrrolizidine alkaloids and quinolones in different honey types. A simple and novel analytical method employing liquid chromatography coupled to high-resolution mass spectrometry using a quadrupole-time-of-flight hybrid analyzer (Q-TOF) was developed and validated. No quinolone residues were detected in 80 honey samples, while pyrrolizidine alkaloids were found in 49% of the investigated samples. © 2021, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.