ISO 17375:2006

Overview - ISO 17375:2006 and aflatoxin B1 testing in feed

ISO 17375:2006 specifies a validated laboratory method for the determination of aflatoxin B1 in animal feeding stuffs. The method uses reverse-phase high-performance liquid chromatography (RP‑HPLC) with post‑column derivatization (PCD) and fluorescence detection. It is applicable to feed matrices with a fat content up to 50%, and the demonstrated limit of quantification (LOQ) is better than 0.5 µg/kg for a signal‑to‑noise ratio of 6.

Key topics and technical requirements

• Analytical principle: extraction with acetone/water, immunoaffinity column (IAC) cleanup, RP‑HPLC separation, PCD via bromination, fluorescence detection.
• Immunoaffinity columns (IACs): antibodies specific to aflatoxin B1; minimum capacity ≥ 40 ng; recovery ≥ 80% for 0.25 ng spike (in acetone/water).
• Post‑column derivatization options:
  • Electrochemically generated bromine, or
  • Chemical reagent pyridinium hydrobromide perbromide (PBPB).
• Mobile phases: defined water/acetonitrile/methanol mixtures; one option (for electrochemical PCD) contains potassium bromide and nitric acid.
• Detection: fluorescence detector (recommended Ex ≈ 365 nm, Em ≈ 435 nm).
• Calibration and quantification: prepared aflatoxin B1 stock and working solutions, procedures for calibration curves and two calibration options (A and B).
• Performance and quality: method precision established by interlaboratory testing; repeatability and reproducibility parameters defined in the standard.
• Safety and good laboratory practice: use of toxic, flammable solvents (acetone, methanol, acetonitrile); aflatoxins are carcinogenic and light‑sensitive; acid‑washed glassware recommended to prevent analyte loss.

Practical applications and who uses ISO 17375

• Feed testing laboratories performing routine surveillance and compliance testing for aflatoxin B1 in animal feed.
• Regulatory authorities and food safety agencies that require standardized, validated methods for enforcement and monitoring.
• Feed manufacturers and quality control teams ensuring raw materials and finished feeds meet safety specifications.
• Contract analytical labs and research groups studying mycotoxin contamination, mitigation, and risk assessment. Benefits include a reproducible HPLC‑based protocol, clear cleanup (IAC) and derivatization options, and an LOQ suitable for low‑level monitoring of aflatoxin B1.

Related standards and references

• ISO 3696:1987 (laboratory water quality) is referenced for water used in reagents and mobile phases.
• Prepared by ISO/TC 34 (Food products), Subcommittee SC 10 (Animal feeding stuffs).

Keywords: ISO 17375, aflatoxin B1, animal feeding stuffs, HPLC, post‑column derivatization, immunoaffinity column, PBPB, electrochemical bromination, fluorescence detection, feed testing.