Ergovaline is produced by the endophyte Neotyphodium coenophialum (Morgan-Jones and Gams) in tall fescue (Schedonorus arundinacea (Schreb.) Dumort. = Festuca arundinacea Schreb.) and is blamed for a multitude of costly livestock disorders. Testing of pastures is common in both research and on farm situations. Since ergovaline is known to be unstable and affected by many variables, the objective of this study was to determine the effect of sample handling and storage on the stability of this compound. Homogeneous milled tall fescue sub-samples were analyzed for ergovaline concentration using HPLC after a range of sample handling procedures or storage. Ergovaline was unstable in milled material after 24 hours in storage, regardless of temperature. The decrease in ergovaline after 24 hours ranged from 17 to 60%. These results show that tall fescue sample handling and storage have a significant effect on ergovaline concentrations. In conclusion, accurate laboratory analysis of ergovaline content may require that samples be transported immediately to the laboratory on ice for immediate analysis. Most laboratories are not equipped for same day analysis, therefore researchers and producers should acknowledge that laboratory ergovaline results may be lower than the actual content in the field.