Simplified Sample Preparation for HPLC Analysis of Ergovaline in Tall Fescue

Abstract

Increased interest in the endophytic fungus, Acremonium coenophialum Morgan‐Jones & Gams, and its symbiotic inhabitance of tall fescue, Festuca arundinacea Schreb., has created the need for rapid and reliable ergovaline analysis in research and in extension service (for diagnosis). A simplified sample preparation method provides faster and more reproducible results for ergovaline quanitation in tall fescue. The method involves extraction with alkaline chloroform, purification with a silica gel solid‐phase chromatography column, and direct ergovaline analysis of methanol eluants by high performance liquid chromatography (HPLC) with fluorescence detection. When compared with a previous method specific for tall fescue vegetative tissue, the simplified method reduced variability and increased recoveries of internal standard from 83 to 94% and of ergovaline from 82 to 93%. This method when used with automated HPLC equipment, permits analysis of 60 to 80 samples per day. Twenty tall fescue samples were analyzed by two separate laboratories using the simplified method; ergovaline results from each were regressed against the other. Ninety‐seven percent of the variability from one laboratory could be accounted for by analysis of the samples by the other, suggesting that the simplified method is highly reproducible.