Straightforward Determination of Pyrrolizidine Alkaloids in Honey through Simplified Methanol Extraction (QuPPE) and LC-MS/MS Modes

[Image: see text] Contamination of honey with toxic pyrrolizidine alkaloids (PAs) and their N-oxides (PANOs) was assessed by dilution with acidic methanol and analysis through liquid chromatography (LC) coupled with tandem mass spectrometry (MS/MS) in three different modes. The hybrid linear trap/triple quadrupole (LC-QTRAP) instrument was used in precursor ion scan (PIS), enhanced product ion scan (EPI), and multiple reaction monitoring (MRM) mode. The parent ions from ions at m/z 120 or 138 amus, characteristic of all the toxic PAs and PANOs in the sample were first scanned by PIS. Then, the presence of each contaminant at specific retention times through its MS(2) spectrum was confirmed by EPI. Finally, they were quantified in the MRM mode. The method was validated: recoveries 86–111%, relative standard deviation (RSD) <20%, at 20 and 40 μg/kg, except retrorsine, which showed a RSD of 30% at 20 μg/kg. Honey samples were analyzed and five of them showed levels of 40 μg/kg for the sum (PAs + PANOs). This approach allows the simultaneous determination of PAs and PANOs in honey, even if their chemical standards are not available.