Ultramarine blue nanoparticles as a label for immunochromatographic on-site determination of ractopamine.

A competitive immunochromatographic assay (ICA) is presented and used for on-site determination of ractopamine (RAC). Ultramarine blue nanoparticles were directly separated from ultramarine blue industrial products by centrifugation (< 10,000 rpm and > 4000 rpm) and used as visible labels in ICAs. The ultramarine blue nanoparticles were coated by polyacrylic acid (PAA), which provides carboxyl groups on the surface of ultramarine blue nanoparticles. An anti-RAC monoclonal antibody (mAb) was covalently immobilized on the carboxyl-modified ultramarine blue nanoparticle surface via diimide-activated conjugation between the carboxyl groups on the ultramarine blue nanoparticle surface and the amino groups of the antibodies. RAC and BSA-modified RAC competitively bind to the anti-RAC mAb on the ultramarine blue nanoparticles. The blue band in the test line is generated by the accumulation of ultramarine blue nanoparticles and is negatively associated with the RAC content. Under optimal conditions, the visual limit of detection (vLOD) of this ICA for RAC is 2.0 ng mL, 2.0 ng mL, and 1.0 ng mL in phosphate-buffered saline (PBS), feed samples, and pork samples, respectively. The ultramarine blue nanoparticle-based ICA also shows no cross activity with salbutamol, clorprenaline, clenbuterol, or terbutaline. Graphical abstract Schematic representation of the ultramarine blue nanoparticles immunochromatographic assay for detection of ractopamine (RAC) based on competitive method. The ultramarine blue nanoparticles were screened from commercial ultramarine pigments for the first time and used to detect ractopamine.