Determination of Aflatoxin B1 in Grains by Aptamer Affinity Column Enrichment and Purification Coupled with High Performance Liquid Chromatography Detection

Abstract

Aflatoxin B1 (AFB1) is a highly teratogenic and carcinogenic secondary metabolite produced by Aspergillus. It is commonly detected in agricultural products such as cereals, peanuts, corn, and feed. Grains have a complex composition. These complex components severely interfere with the effective extraction and separation of AFB1, and also cause problems such as matrix interference and instrument damage, thus posing a great challenge in the accurate analysis of AFB1. In this study, an aptamer affinity column for AFB1 analysis (AFB1-AAC) was prepared for the enrichment and purification of AFB1 from grain samples. AFB1-AAC with an AFB1-specific aptamer as the recognition element exhibited high affinity and specificity for AFB1. Grain samples were enriched and purified by AFB1-AAC, and subsequently analyzed by high performance liquid chromatography with post-column photochemical derivatization-fluorescence detection (HPLC-PCD-FLD). The average recoveries of AFB1 ranged from 88.7% to 99.1%, with relative standard deviations (RSDs) of 1.4–5.6% (n = 3) at the spiked levels of 5.0–20.0 μg kg−1. The limit of detection (LOD) for AFB1 (0.02 μg kg−1) was much below the maximum residue limits (MRLs) for AFB1. This novel method can be applied to the determination of AFB1 residues in peanut, corn, and rice.