Preparation of an Immunoaffinity Column with Amino-Silica Gel Microparticles and Its Application in Sample Cleanup for Aflatoxin Detection in Agri-Products

Abstract

This study established an immunoaffinity column for selective extraction of aflatoxins in agri-products. Specifically, the immunoaffinity column was developed by covalently coupling monoclonal antibody 1C11 against aflatoxins to amino-silica gel microparticles and then packing these into a cartridge. The extraction conditions were thoroughly optimized in terms of loading, washing and eluting solutions. Under the optimal conditions, the immunoaffinity column had a capacity of 200 ng of aflatoxins. The detection limits (S/N = 3) for aflatoxin G1, B1, G2 and B2 were 0.03, 0.07, 0.05 and 0.09 μg·kg−1, and the corresponding quantification limits (S/N = 10) were 0.10, 0.25, 0.18 and 0.30 μg·kg−1, respectively. The recoveries of aflatoxins in samples were 90.1%–104.4% and RSDs were <4.4%. The developed method was further applied to the determination of aflatoxins in peanut, vegetable oil and tea samples, and the results indicated that peanut (26.9%), vegetable oils (28.0%) and tea (5.3%) samples were contaminated with aflatoxins, with levels ranging from 0.49 to 20.79 μg·kg−1.

Keywords

• aflatoxins
• immunoaffinity column
• amino-silica gel
• microparticle conjugate
• agri-products
• HPLC