Development of an Analytical Method for Analyzing Pyrrolizidine Alkaloids in Different Groups of Food by UPLC-MS/MS

Suspected nontargeted pyrrolizidine alkaloids (PAs), without analytical reference standard, were observed and interfered with the determination of targeted PAs in complex food matrices, especially for spices samples. Selectivity and applicability of multiple reaction monitoring (MRM) transitions, multistage fragmentation (MS3), and MRM with differential ion mobility spectrometry (DMS) for eliminating false positive identifications were evaluated. Afterward, a selective and sensitive LC-MS/MS method for the determination of 15 PAs and 13 PA N-oxides in foodstuffs was developed. The sample preparation and cleanup are applicable to a wide range of foodstuffs, including cereal products, dairy products, meat, eggs, honey, tea infusion, and spices. Freezing-out of the raw extract and the water/acetonitrile washing steps in a solid phase extraction was found to efficiently remove complex matrices. The method was validated at 0.05 μg kg^–1 for general food and 0.5 μg kg^–1 for spices, with reference to the Eurachem Guide. The estimated limit of quantifications of different PAs was in the range of 0.010–0.087 μg kg^–1 for general food and 0.04–0.76 μg kg^–1 for spices. Isotopically labeled PAs were used as internal standards to correct the variation of PAs/PANs performance in different food commodities. Matrix effects observed in complex food matrices could be reduced by solvent dilution. Recoveries of PAs and PA N-oxides were all seen within 50–120%