Optimization and Validation of ELISA for Aflatoxin B1 Detection in Fermented Forages and Feeds

Abstract

Enzyme-coupled immunosorbent assays (ELISA) methods are usually validated only for homogenous matrixes like corn and wheat. More complex materials like fermented forages and mixed feed are not targeted for mycotoxin measurement. The low number of ELISA methods found in the literature neither contained the pH set for fermented forages nor dealt with the setting of the matrix:solvent ratio. The sample preparation of these matrixes needs to be optimized and validated for aflatoxin B1 analysis from fermented forages (corn silage and rye haylage) and mixed feed for Romer AgraQuant® Aflatoxin B1 ELISA (Romer Labs, Austria). Drying and pH adjustment of fermented forages had high importance before mycotoxin extraction. Because of the matrix swelling, the 1: 5 ratio of the sample/extraction solute should have been increased to 1: 8 to gain the highest aflatoxin B1 recovery. The accuracy and repeatability of the analysis were tested and found to be suitable for further application.