Targeted LC–MS/MS Analysis of 11 Ranunculaceae Phytotoxins in Honey: Method Development, Validation, and Application to Real Samples

The consumption of honey contaminated with phytotoxins, particularly from Ranunculaceae plants, is a global public health concern due to their widespread distribution and diverse toxic compounds. This study developed a UPLC-QQQ-MS method to simultaneously quantify 11 phytotoxins of Ranunculaceae family in honey. Analytical (mobile phase) and pre-treatment (extraction solvent and cleanup column) procedures were optimized, identifying Methanol–Water (added 0.1% formic acid and 5 mM ammonium formate), acetonitrile, and SCX column as optimal. Method validation demonstrated negligible matrix effects, good linearity (R 2  > 0.995), good recoveries (81.38–116.85%), and high precision (RSD ≤ 8.63%) for 25–500 ng/mL ranges. LODs and LOQs were 0.02–0.15 µg/kg and 0.07–0.50 µg/kg, respectively (excluding anemonin). Retrospective analysis of 50 honey samples detected magnoflorine, talatisamine, hypaconitine, senecionine, and protopine at positive rates of 2%, 2%, 2%, 8%, and 10%, respectively. Thus, this work provides important technical supports for the risk assessment of phytotoxins in honey.