Determination of ergovaline in endophyted seeds by high performance thin layer chromatography (HPTLC)

High performance thin layer chromatography (HPTLC) is used to rapidly determine the presence and amount of ergovalin in seeds of Graminaceae infested with Neotyphodium Coenophialum. Quinine is used as an internal standard. The mycotoxin is solubilized under acid conditions and extracted with chloroform under alkaline conditions. Samples are sprayed on silica gel plates and detected by fluorescence with lambda (1) (excitation) set at 318 nm and lambda (2) (emission) > 400 nm. The method developed is specific for endophyte-infested seeds, linear from 3 to 12 mug/g and is both repeatable and reproducible. The limit of detection is 0.5 mug/g and the limit of determination (quantification) is 3.0 mug/g. This technique was applied to several batches of seeds obtained from pastures that may or may not be infested with the fungus. Results were compared with results obtained using a standard High Performance Liquid Chromatography (HPLC) method.