An HPLC method for the detection of ergot in ground and pelleted feeds.

A high performance liquid chromatography (HPLC) method is described for detecting ergot in ground or pelleted forages and grains. Samples were extracted with alkaline chloroform, filtered, and applied to silica gel/organic binder cleanup columns. Following elution of pigments with acetone: chloroform, ergopeptine alkaloids were eluted with methanol and analyzed by HPLC with fluorescence detection. Average recovery of ergotamine, the major ergopeptine alkaloid produced by Claviceps, was 93%, with a relative standard deviation of 4.9%. The detection limit of ergotamine was approximately 50 ppb in all feedstuffs. Confirmation of ergopeptine alkaloids was accomplished by treating the parent ergopeptine alkaloids with 0.2% acetic acid to produce their -inine isomers and reexamining by HPLC with fluorescence detection or silica gel/organic binder column cleanup in combination with tandem mass spectroscopy. The method described is a valid alternative to microscopic inspection for detecting ergot contamination in ground or pelleted feedstuffs.