Novel paper-based colorimetric immunoassay (PCI) for sensitive and specific detection of salbutamol residues in flesh of swine and urine using Ag3 PO4 /Ag nanocomposite as label

Abstract

Salbutamol (SAL) can cause potential hazards to human health and its use as a growth promoter in meat-producing animals is illegal. This work reports a novel approach for competitive paper-based colorimetric immunoassay (PCI) using the Ag₃ PO₄ /Ag nanocomposite as label for sensitive and specific determination of SAL in flesh of swine and urine. The Ag₃ PO₄ /Ag nanocomposite was synthesized by a one-step chemical bath method, which could instantly oxidize a chromogenic substrate for the color development under acidic conditions without the participation of H₂ O₂ . This approach provides high affinity between the Ag₃ PO₄ /Ag nanocomposite and the substrate (with the Michaelis-Menten constant of 0.44 mM). In addition, the fabrication process of the PCI was simple and cost-effective. Particularly, the novel PCI also exhibits simplicity and cost-effectiveness of the fabrication process through a simple wax screen-printing, which requires inexpensive equipment and material including a screen, wax, a squeegee, and a hair dryer. Under optimal conditions, the competitive PCI exhibited a linearity range of 0.025 to 1.00 µg/L. The developed approach offers advantages over the conventional ELISA for the purpose of routine use because it requires a shorter incubation time (<1 hr), significantly small volumes of reagents and samples (<100 µL each), and an inexpensive consumer-grade digital camera coupled with a simple gray-scale transformation of the RGB (Red Green Blue) color image for the purpose of quantification of the detection. PRACTICAL APPLICATION: Salbutamol (SAL) can cause potential hazards to human health and the use of which as growth promoter in meat-producing animals is illegal. This work introduces a novel approach for competitive immunoassay on paper-based colorimetric immunoassay using the Ag₃ PO₄ /Ag nanocomposite as the label (instead of using natural enzyme) for low-cost, sensitive, and specific determination of SAL residues at low level in flesh of swine and urine samples. The proposed approach offers advantages over the conventional ELISA for the purpose of routine use.

Keywords: Ag3PO4/Ag nanocomposite; Red Green Blue (RGB) system; paper-based colorimetric immunoassay (PCI); salbutamol; screen-printed technique; swine.