Pyrrolizidine alkaloids in natural and experimental grass silages and implications for feed safety

Hepatotoxic 1,2-dehydro-pyrrolizidine alkaloids (PA) and their N-oxides (PANO) in feed are a potential threat for animal and human health. However, their risk assessment in preserved animal forage is difficult due to data gaps regarding their occurrence in field samples and contradictory results regarding their behavior during the ensilage process. In this study, 115 samples of grass silage originating from different districts in Bavaria (Germany) were analyzed for their PA and PANO contents. A sensitive LC-ESI-MS/MS method for the detection of 10 PA and 4 PANO was developed including a clean-up of the aqueous acidic extract by cation-exchange cartridges. The mean recoveries were between 70% for seneciphylline-. N-oxide and 104% for senecionine-. N-oxide. The limits of detection ranged between 0.14. μg senkirkine/kg dry matter (DM) and 1.3. μg retrorsine-. N-oxide or monocrotaline-. N-oxide/kg DM. Eighteen percent of the samples contained one or more of the analyzed compounds. The highest observed value, calculated as the sum of seneciphylline and senecionine, was 30. μg/kg DM. Other samples contained senecionine-. N-oxide, seneciphylline-. N-oxide, lycopsamine, intermedine, echimidine, and heliotrine in lower amounts. An accompanying laboratory scale ensilage trial showed a compound-dependent transformation of PANO during the fermentation process while PA-amounts remained stable or even increased. Impacts on safety of ensiled animal feed are discussed with regard to animal health.