Overview
EN ISO 16050:2011 (identical to ISO 16050:2003) specifies a validated analytical procedure for the determination of aflatoxin B1 and the total content of aflatoxins B1, B2, G1 and G2 in cereals, nuts and derived products. The method uses reverse‑phase high‑performance liquid chromatography (HPLC) with immunoaffinity column (IA) clean‑up and post‑column derivatization plus fluorescence detection. The standard sets a limit of quantification (LOQ) of 8 µg/kg for aflatoxin B1 and for the sum of B1+B2+G1+G2 and includes validated performance data for specific matrices.
Key topics and technical requirements
- Analytical principle: extraction with methanol/water, IA column affinity purification, elution and quantification by reverse‑phase HPLC with fluorescence detection after post‑column derivatization.
- Clean‑up: immunoaffinity columns specific to aflatoxins B1, B2, G1, G2.
- Derivatization reagents: iodine solution or pyridinium hydrobromide perbromide (PBPB) as post‑column oxidizing reagents (document lists preparation details).
- Typical reagents and solvents: methanol/water extraction (7:3 v/v), mobile phase (water:acetonitrile:methanol 3:1:1), toluene/acetonitrile mixture for stock standards (as described in the standard).
- Quantification and calibration: use of aflatoxin standard solutions, UV spectrometric determination of stock concentrations, and calibration graphs for HPLC response.
- Performance and validation: LOQ = 8 µg/kg; method validated for maize (total aflatoxins ~24.5 µg/kg), peanut butter (~8.4 µg/kg), raw peanuts (~16 µg/kg). Method applicability extended to oilseed products, dried fruits and derived products.
- Safety and handling: aflatoxins are carcinogenic - the standard highlights light protection, fume hood use for solvents (toluene), and safe laboratory practice.
Applications
- Regulatory compliance testing for aflatoxin limits in cereals, nuts and related food products.
- Food industry quality control: incoming raw material screening, product release testing and supplier verification.
- Import/export surveillance: testing consignments for aflatoxin contamination to meet trade and sanitary requirements.
- Analytical laboratories and research performing quantitative aflatoxin analysis or method validation.
Who should use this standard
- Accredited food testing laboratories and quality assurance teams in cereal, nut and oilseed processing.
- Public health and food safety authorities, customs laboratories, and contract testing providers.
- Method developers and researchers working on mycotoxin analysis.
Related standards
- Supersedes EN 12955:1999 (withdrawn).
- References ISO 3696 (water for analytical use) and other ISO/CEN food analysis guidance.
Keywords: EN ISO 16050:2011, ISO 16050, aflatoxin B1, total aflatoxins, HPLC method, immunoaffinity column, post‑column derivatization, cereals, nuts, food safety testing.