DIN EN 15851:2010-07

This European Standard specifies a method for the determination of aflatoxin B1 in baby food by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection. This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 0,07 µg/kg to 0,18 µg/kg. A test portion is extracted with a mixture of methanol and water. The extract is filtered, diluted with phosphate buffered saline (PBS) to a specified solvent concentration, and applied to an immunoaffinity column containing antibodies specific to aflatoxin B1. Aflatoxin B1 is purified and concentrated on the column and removed from the antibodies using methanol as eluent. Aflatoxin B1 is quantified by reverse-phase high performance liquid chromatography (RP-HPLC) with post column derivatization (PCD) involving bromination followed by fluorescence detection. The post column derivatization is achieved with either electrochemically generated bromine or with pyridinium hydrobromide perbromide (PBPB).