EN ISO 14501:2021 specifies a standardized analytical method for the determination of aflatoxin M1 in milk and milk powder. The method combines sample clean‑up by immunoaffinity chromatography (IAC) with quantitative determination by high‑performance liquid chromatography (HPLC). It is applicable to whole, low‑fat and skimmed liquid milks and to whole, low‑fat and skimmed milk powders. Safety notes in the standard warn that aflatoxins are carcinogenic and that standard solutions must be protected from light.
Key performance characteristics declared in the scope:
- Validation level: 0.08 µg/kg for whole milk powder (equivalent to 0.008 µg/L for reconstituted liquid milk)
- Limit of detection (LOD): 0.05 µg/kg (milk powder) and 0.005 µg/kg (liquid milk)
- Limit of quantification (LOQ): 0.1 µg/kg (milk powder) and 0.01 µg/kg (liquid milk)
Key topics and technical requirements
- Principle: Extraction/preparation of milk samples, IAC clean‑up to selectively bind aflatoxin M1, elution and concentration of the analyte, and HPLC analysis with appropriate detection.
- Sample preparation: Detailed procedures for milk and milk powder reconstitution and extraction to ensure representative testing.
- Immunoaffinity column preparation: Instructions for conditioning and use of IAC for specific aflatoxin M1 capture.
- HPLC conditions: Pump settings, chromatographic performance criteria, calibration curve construction and injection schemes required for reliable quantification.
- Calculations and expression of results: How to report aflatoxin M1 concentrations for different matrices (milk and powder).
- Precision and performance: Interlaboratory trial results, repeatability and reproducibility information to support method reliability.
- Safety: Explicit cautions about carcinogenicity of aflatoxins and protection of standards and samples from light.
Applications
- Routine dairy product testing for food safety monitoring and regulatory compliance.
- Quality control in dairy processing and powdered milk production.
- Surveillance testing for aflatoxin M1 in supply chains, imports/exports and raw milk screening.
- Reference method for laboratories seeking validated HPLC + IAC protocols for aflatoxin M1 analysis.
Who should use this standard
- Commercial and public food testing laboratories
- Dairy industry quality assurance teams
- National and regional food safety authorities and regulators
- Laboratories performing method validation, proficiency testing and conformity assessment for dairy products
Related standards and notes
- This document is the third edition, published jointly by ISO and IDF and adopted by CEN as EN ISO 14501:2021, superseding EN ISO 14501:2007.
- Users should consult national/regulatory limits for aflatoxin M1 and adopt laboratory safety protocols when handling aflatoxins.
Keywords: EN ISO 14501:2021, aflatoxin M1, milk testing, milk powder, immunoaffinity chromatography, HPLC, limit of detection, limit of quantification, dairy laboratory, food safety.