Determination of trace aflatoxin M1 (AFM1) residue in milk by an immunochromatographic assay based on (PEI/PSS)4 red silica nanoparticles

Aflatoxin M1 (AFM1) residues in milk pose a major threat to human health, so there is an urgent need for a simple, rapid, and sensitive method for the determination of trace AFM1 in milk. In this study, a competitive immunochromatographic assay (ICA), using visual (PEI/PSS)(4) red silica nanoparticles (SiNPs) as signal amplification probes, was used for the highly sensitive detection of AFM1. The (PEI/PSS)(4) red SiNPs were used to label AFM1 monoclonal antibody (mAb) to prepare ICA for the detection of AFM1. After exploring the optimal conditions of mAb and immunoprobe dosage conditions, the lowest visual detection limit (VDL) of AFM1 in phosphate-buffered saline with Tween 20 (PBST, 10 mM, pH 7.4, containing 1% BSA, 3% sucrose, 1% trehalose, and 0.5% Tween 20) can reach 0.1 pg/mL. The intuitive visually visible value of AFM1 in both PBST and milk was 10 pg/mL. The results showed that the immunochromatographic system based on high chroma color (PEI/PSS)(4) red SiNPs has high sensitivity and broad application prospects for the detection of trace AFM1 residues in milk. The high chroma (PEI/PSS)(4) red SiNPs are expected to be a convenient biomarker for improving the sensitivity of immune chromatography bands. The schematic diagram shows the detection principle. In this work, in the competitive experiment, (PEI/PSS)(4) red SiNPs were selected as visual labeling materials, and the specific antibody combined with the labeled material was selected as an immune probe. The AFM1-BSA antigen coupled with the macromolecular BSA was fixed on the T line of the nitrocellulose (NC) membrane. The AFM1 in sample solution competes with AFM1-BSA for the specific binding site of immune probe. The detection sensitivity of this method for AFM1 is obtained by judging the change of the red signal intensity produced by the positive sample, compared with the color at the T line of the negative sample.