Determination of aflatoxins in animal feeds by HPLC with multifunctional column clean-up

A reversed-phase HPLC method with fluorescence detection for the determination of the aflatoxins B1, B2, G1 and G2 in 42 animal feeds, comprising corn (16), soya bean meal (8), mixed meal (13), sunflower, wheat, canola, palm kernel, copra meals (1 each) was carried out. The samples were first extracted using acetonitrile:water (9:1), and was further cleaned-up using a multifunctional column. Optimum conditions for the extraction and chromatographic separation were investigated. By adopting an isocratic chromatographic system using a mobile phase comprising acetonitrile:methanol:water (8:27:65, v/v/v), the separation of the four aflatoxins was possible within 30 min. Recoveries for aflatoxins B1, B2, G1 and G2 were 98 ± 0.7%, 95 ± 1.0%, 94 ± 3.6% and 97 ± 4.3%, respectively. The results show that eight samples (19%) were contaminated with aflatoxins, ranging from 6.5 to 101.9 ng g−1. Total aflatoxin levels in three samples exceed the legal limits of many countries of 20 ng g−1.