Determination of pyrrolizidine alkaloids in Hungarian honeys by a QuEChERS-based HPLC-MS/MS method and human health risk assessment

Pyrrolizidine alkaloids (PAs) are frequently occurring phytotoxins in honeys, however, representative data are limited to a few countries. In the current research, a citrate-buffered QuEChERS-based multicomponent HPLC-MS/MS method was validated for honey matrix, using a triple quadrupole mass analyzer. The greenness of the methodology was assessed using the Analytical GREEnness (AGREE) metric approach. Recovery and precision were evaluated at four concentration levels (1.5, 4.0, 8.0 and 20.0 µg/kg). Satisfactory results were obtained for LOD (0.13-2.50 µg/kg), LOQ (1.50-7.50 µg/kg), linearity (0.984<R 2 <0.999), and intra-day repeatability (<20%). Considerable matrix suppression was observed for certain analytes, which necessities matrix matched calibration. Recoveries exceeded 120% for three analytes, which were therefore excluded from validation. The method was applied to 50 Hungarian honeys from various botanical and geographical origins. Echimidine was detected in the majority of samples, mostly in trace amounts. Lycopsamine-type and senecionine-type PAs also occurred frequently at varying concentrations. Palynological evaluation revealed the presence of pollen from PA-producing plants in monofloral honeys, but pollen composition was not closely related to PA contamination. Results suggest that average honey consumption does not pose a significant food safety risk to adults. However, regular high intake of contaminated honeys may be a concern. • A HPLC-MS/MS method was validated for 18 of 21 PAs, including 4 co-eluting isomers • Hungarian honeys (n=50) were characterized based on their PA composition • Palynology poorly predicts PA in honey due to species over-/under-representation • Regular high honey consumption may result in concerning PA exposure