Enhancing pyrrolizidine alkaloid separation and detection: LC-MS/MS method development and integration of ion mobility spectrometry into the LC-HRMS workflow

Pyrrolizidine alkaloids (PAs) are plant toxins occurring in different foodstuffs, including teas, herbal infusions and species. Additionally, PAs may be transferred to honey and pollen when honeybees come into contact with contaminated plants. Due to their adverse effect, PAs occurrence in food must be controlled to ensure public health. Nevertheless, the presence of numeours PA epimers complicates their chromatographic separation and detection. In this regard, a method using liquid chromatography coupled with tandem mass spectrosmetry (LC-MS/MS) has been developed allowing the separation of 31 out of the 35 regulated PAs, which was sucessfully validated in different food matrices such as herbal infusions, spices and honey. Afterwards, travelling wave ion mobility spectrometry hyphenated with quadrupole time-of-flight mass spectrometry (TWIMS-QTOF) was evaluated to improve the analytical performance of PAs determination. Thus, collision cross section (CCS) values of PAs have been therefore obtained for the first time. The CCS library for PAs was also compared with predicted values by machine learning and with those meassured in real food matrices (bias <2 %). In addition, an in-house library was used in the suspect screening of PAs to complement the targeted analysis of the studied samples, all of which tested positive for several PAs.