Comparison of two extraction methods for ergosterol determination in vegetal feeds

Abstract

Ergosterol is the principal sterol of fungi in which it plays an essential role in cell membrane and other cellular constituents. This sterol is considered as a good marker of fungal contamination and of mycotoxin production. After validation of ergosterol quantification by HPLC-UV system (linearity range: 0.2 to 20.0 mg/ml, repeatability: 3.27%, between day precision: 4.75%), 2 extraction methods of ergosterol from 3 vegetal matrixes (maize, barley and wheat) were compared: the first one, normalized by the AFNOR, is based on solid phase extraction (SPE), while the other is based on liquid/liquid extraction (LLE). The LLE procedure allowed ergosterol extraction gains of around 20% for high initial sterol contents (3 to 5 mg/kg) in naturally contaminated matrixes or in spiked samples, and of 86% for low initial sterol contents (1-2 mg/kg) in maize. Moreover, the precision of ergosterol determination was comparable for the 2 methods even if it was slightly lower using LLE and was more affected by the initial ergosterol contents in vegetal matrix than by its nature. These results suggest that ergosterol contents in vegetal feeds would be underestimated with the official method (SPE) and emphasize the importance of the extraction step.