Development and application of solvent-free extraction for the detection of aflatoxin M1 in dairy products by enzyme immunoassay

The official methods for the quantification of aflatoxin M1 in dairy products (cheese and yogurt) include extraction into dichloromethane or chloroform, evaporation of the solvent, partitioning of the reconstituted residue with hexane, and subsequent analysis. To secure a rapid and inexpensive screen for aflatoxin M1 contamination, a sensitive competitive ELISA, using a rabbit polyclonal antibody, was developed for measuring aflatoxin M1 in milk and used in a comparative study for measuring the extraction efficiency of aflatoxin M1 in aqueous or organic solvent buffers using yogurt samples. An aqueous sodium citrate solution was found to be suitable for extracting aflatoxin M1, thus eliminating the need for organic solvents. The citrate extraction proved to be efficient (recovery ranged from 70 to 124%) in fortified samples of very different kinds of dairy products, including yogurt and six types of cheese. Fourteen yogurt and cheese samples were extracted with citrate solution and analyzed by ELISA. A good correlation was observed (y=0.95x-0.59, r2=0.98) when the data were compared with those obtained through the official method, across a wide range of aflatoxin M1 contaminations (10-200 ng/kg).     

Selective determination of melamine in aqueous medium by molecularly imprinted solid phase extraction

A molecularly imprinted polymer able to recognize melamine in partially aqueous medium was synthesized using methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linking agent. The bound specificity and selectivity of the obtained material were verified by performing binding experiments with melamine and its structural analogue, 2,4,6-trimethoxy-1,3,5-triazine, respectively, using different aqueous binding media. Finally, the ability of MIP to selectively extract melamine from two real samples, a food supplement and a freeze-dried meat sample, was demonstrated.