Overview
ISO 17372:2008 specifies a validated laboratory method for the determination of zearalenone in animal feeding stuffs using immunoaffinity column (IAC) chromatography followed by high performance liquid chromatography (HPLC) with fluorescence detection. The standard applies to a wide range of feed matrices (barley, corn, oats, rye, wheat, soybean and canola meals, corn gluten, dried distillers’ grains, lentils, sugar beet pulp, etc.) and defines a practical limit of quantification (LOQ) of 0.05 mg/kg (50 µg/kg); lower LOQs are possible with appropriate in‑house validation.
Key topics and technical requirements
- Scope and matrices: Designed for common feed ingredients and complete animal feeding stuffs.
- Sample preparation: Grinding to ≤1 mm (recommendation for finer particle size where practical) and representative sampling per ISO 6498 guidance.
- Extraction: Organic extraction using acetonitrile‑based solvent (extraction solvent φ(CH3CN)=90 %) with salt (NaCl) followed by filtration.
- IAC cleanup: Purification on zearalenone‑specific immunoaffinity columns to concentrate and remove interferences; instructions for handling pigmented or problematic matrices (use of methanol washes, filter/frit arrangements).
- HPLC analysis: Reverse‑phase HPLC with fluorescence detection (excitation wavelengths including 236 nm and 274 nm, emission at ~418–440 nm). Mobile phase and standard preparation procedures are specified to generate calibration curves.
- Quality control: Use of spiked control samples (e.g., 0.10 mg/kg) with recommended recovery ≥85 % and preparation of stock and working standards; evaporative concentration of IAC eluates to improve sensitivity.
- Confirmation options: Wavelength ratioing, normal‑phase HPLC, or diode‑array detection for suspect positives.
- Documentation: Calculation of results, precision statements, and test reporting requirements are included.
Applications and practical value
- Routine mycotoxin monitoring in feed manufacturing, grain handling, and feed ingredient supply chains.
- Regulatory compliance testing and surveillance by laboratories supporting veterinary public health and animal nutrition.
- Quality control in feed mills and grain testing laboratories to protect animal health and prevent carry‑over of mycotoxins into animal products.
- Research and method validation efforts aiming to push LOQ lower with additional laboratory validation.
Who should use this standard
- Accredited analytical laboratories performing feed mycotoxin testing.
- Feed manufacturers, grain exporters, and quality assurance teams.
- Regulatory agencies and research institutions engaged in feed safety and risk assessment.
Related standards
- ISO 6498 (preparation of test samples), ISO 3696 (water for analytical use), ISO 565 (sieves) - all referenced for sampling, glassware and sample preparation practices.
Keywords: ISO 17372:2008, zearalenone, immunoaffinity column, HPLC, animal feeding stuffs, mycotoxin analysis, limit of quantification, feed testing.
Open preview (PDF)Animal feeding stuffs — Determination of zearalenone by immunoaffinity column chromatography and high performance liquid chromatography