Polystyrene microsphere-mediated optical sensing strategy for ultrasensitive determination of aflatoxin M₁ in milk

Aflatoxin M₁ (AFM₁) contamination poses a serious threat to human health globally. Hence, it is necessary to develop reliable and ultrasensitive methods for the determination of AFM₁ residue in food products at low levels. In this study, a novel polystyrene microsphere-mediated optical sensing (PSM-OS) strategy was constructed to solve the problems of low sensitivity and susceptibility to interference from the matrix in AFM₁ determination. Polystyrene (PS) microspheres have the advantages of low cost, high stability, and controllable particle size. They can be useful optical signal probes for qualitative and quantitative analyses attributed to the fact that they have strong ultraviolet-visible (UV-vis) characteristic absorption peaks. Briefly, magnetic nanoparticles were modified with the complex of bovine serum protein and AFM₁ (MNP₁₅₀-BSA-AFM₁), and biotinylated antibodies of AFM₁ (AFM₁-Ab-Bio). Meanwhile, PS microspheres were also functionalized with streptavidin (SA-PS₉₅₀). In the presence of AFM₁, a competitive immune reaction was triggered leading to the changes in AFM₁-Ab-Bio concentrations on the surface of MNP₁₅₀-BSA-AFM₁. The complex of MNP₁₅₀-BSA-AFM₁-Ab-Bio binds with SA-PS₉₅₀ to form the immune complexes due to the special binding of biotin and streptavidin. The remaining SA-PS₉₅₀ in the supernatant was determined by UV-Vis spectrophotometer after magnetic separation, which positively correlated with the concentration of AFM₁. This strategy allows for ultrasensitive determination of AFM₁ with limits of detection as low as 3.2 pg/mL. It was also successfully validated for AFM₁ determination in milk samples, and a high consistency was found with the chemiluminescence immunoassay. Overall, the proposed PSM-OS strategy can be used for the rapid, ultrasensitive, and convenient determination of AFM₁, as well as other biochemical analytes.