EN 15792:2009 – Animal feeding stuffs — Determination of zearalenone in animal feed — High performance liquid chromatographic method with fluorescence detection and immunoaffinity column clean-up

1 Scope
This Standard is applicable to the determination of zearalenone in animal feed at concentrations from 30 µg/kg to 3 000 µg/kg.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies: EN ISO 3696, Water for analytical laboratory use – Specification and test methods (ISO 3696:1987)
3 Principle
Zearalenone is extracted from the commodity using organic solvent. The solvent extract is then diluted with phosphate buffered saline to give an aqueous extract which is applied to an immunoaffinity column containing antibodies specific for zearalenone. The analyte is isolated, purified and concentrated on the column and removed from the antibodies with elution solvent. Zearalenone is quantitatively determined by high performance liquid chromatography (HPLC) with fluorescence detection.
4 Reagents
During the analysis, unless otherwise stated, use only reagents of recognised analytical grade and only distilled water or water of grade 1 as defined in EN ISO 3696. Solvents shall be of quality for HPLC analysis.
4.1 Acetonitrile WARNING — Acetonitrile is hazardous and handling shall be carried out inside a fume cupboard. Appropriate safety equipment (lab coat, goggles, gloves) shall be worn.
4.2 Methanol, technical grade WARNING — Methanol is hazardous and handling shall be carried out inside a fume cupboard. Appropriate safety equipment (lab coat, goggles, gloves) shall be worn. Samples shall be blended using an explosion proof blender.
4.3 Methanol, HPLC grade WARNING — Methanol is hazardous and handling shall be carried out inside a fume cupboard. Appropriate safety equipment (lab coat, goggles, gloves) shall be worn. Samples shall be blended using an explosion proof blender.
4.4 Sodium chloride
4.5 Disodium hydrogen orthophosphate
4.6 Potassium dihydrogen phosphate
EN 15792:2009 – Animal feeding stuffs — Determination of zearalenone in animal feed — High performance liquid chromatographic method with fluorescence detection and immunoaffinity column clean-up