Comparative Study of 52734-U-SupelMIP™ Solid-Phase and QuEChERS Extraction Methods for Monitoring 5-Nitroimidazoles in Albanian Honey

The use of 5-nitroimidazoles such as metronidazole, dimetridazole, ronidazole, tinidazole and ipronidazole against Nosema infections raises significant health concerns due to their mutagenic and carcinogenic effects. This study optimized two extraction methods for 5-nitroimidazoles in honey: 52734-U-SupelMIP™ Solid-Phase Extraction and QuEChERS, analyzed by LC-MS/MS technique. The optimized method achieved detection limits of (0.05–0.19 µg/kg) and quantification limits of (0.33–0.46 µg/kg). Linearity was demonstrated (R ²  > 0.99) over (0.3-3.0 µg/kg), with repeatability < 17.24% RSD and recovery rates of (69.98–102.70)%. 52734-U-SupelMIP™ Solid-Phase Extraction showed 35.35% higher extraction efficiency than QuEChERS extraction method. Analysis of 24 Albanian honey samples revealed residues in 12.5% of samples ([0.59-53.00 µg/kg] range). Three samples exceeded the Minimum Required Performance Limit (MRPL) set by Commission Implementing Regulation (EU) 2021/808 of 1 µg/kg, with maximum levels of (53.00 µg/kg) for metronidazole. These findings highlight the urgent need for rigorous nitroimidazole monitoring in honey to protect bee and human health.