Complete compilation of applications for food analysis

Assay of Aflatoxin M1 in Milk Based on Notification Test Methodology, Using Prominence-i and the RF-20AXS Fluorescence Detector

Provide a rapid, sensitive method for quantifying aflatoxin M1 (AFM1) in milk by HPLC–fluorescence, aligned with Japanese Food Safety notices. AFM1, a carcinogenic metabolite of aflatoxin B1, is strictly regulated in milk and dairy. The method achieves an LOQ of 0.1 µg/L, 1/100th of Japan’s regulatory limit, using minimal sample prep and standard fluorescence detection.

• Notification background: AFM1 in milk limited to 0.5 µg/kg. Official HPLC–fluorescence method (confirmation by LC–MS/MS) mandated Jan 2016.
• Sample prep: Milk (homogenized) or raw milk (centrifuged) spiked with AFM1 standard, warmed, filtered, cleaned via AflaStar™ R immunoaffinity column, eluted, dried under N2, reconstituted in 80/20 water/acetonitrile.
• HPLC–FLD conditions: Shim-pack VP-ODS column (150×4.6 mm, 5 µm), 40 °C. Mobile phase H2O/ACN 3/1, 1.0 mL/min. Injection 100 µL. RF-20AXS ex365/em435 nm, cell 25 °C.
• Performance: 0.1–20 µg/L linear range (R2 ≥ 0.9999). Six-replicate LOQ (0.1 µg/L) RSD 3.4 %. Recovery in spiked milk (0.05 µg/kg AFM1; 1/10th regulatory) was 98 %. Trace AFM1 detected in unspiked samples at <1/100th regulatory level by LC–MS/MS.
• Conclusions: Shimadzu Prominence-i and RF-20AXS FLD meet regulatory HPLC–fluorescence requirements for AFM1 screening in milk, providing high sensitivity, precision, and ease of operation.

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