Determination of ergot alkaloids from grains with UPLC-MS/MS

A simple and rapid method for determining six ergot alkaloids and four of their respective epimers was developed for rye and wheat. The analytes were extracted from the sample matrix with ACN/ammonium carbonate solution. The extract was purified with a commercial push-through SPE column (Mycosep 150 Ergot). After concentration and filtration steps, the final separation of the analytes was achieved with ultra-performance LC-MS/MS. The chromatographic separation of the ergot alkaloids was achieved in 4.5 min. The method performance proved satisfactory in the preliminary validation. The calculated LOQs were low ranging from 0.01 to 1.0 microg/kg for wheat and from 0.01 to 10.0 microg/kg for rye. At the concentration levels of 10, 50 and 200 microg/kg, the recoveries were between 80 and 120% in most cases and the within-day repeatability (expressed as RSD) ranged between 1.3 and 13.9%. Despite the cleanup of the samples, some matrix effect was observed in the MS, highlighting the necessity of using matrix-assisted standards. This is the first article to describe the application of the push-through columns and ultra-performance LC in the analysis of ergot alkaloids.