Colorimetric determination of tyrosinase based on in situ silver metallization catalyzed by gold nanoparticles

Gold nanoparticles (AuNPs) catalyze the mild reaction between the weak reducing agent kojic acid (KA) and silver ions (Ag+) to form Au@Ag bimetallic NPs by the combination of the intrinsic catalysis with plasmonic properties This is proposed as a novel optical assay to determine the tyrosinase (TYRase) concentration. The nanoparticles have been characterized by UV-vis spectroscopy, transmission electron microscope (TEM) images, and X-ray photoelectron spectroscopy (XPS). The sensing mechanism is based on the fact that KA binds to TYRase by chelating with dicopper active site of TYRase and the introduction of TYRase restrains the Au@Ag bimetallic NP formation by the precedent binding with KA. A clear color variation from yellow to pink and UV-vis spectral changes are observed at the optimal wavelength of 410 nm. The assay works in the range 0.13~0.73 U mL−1 with a detection limit (LOD) of 0.019 U mL−1. The impact from matrix interfering substances including glucose, uric acid, common oxidases, and amino acids is negligible. The applicability is demonstrated by quantitative determination of TYRase in human serum samples with 74 to 89% recovery and RSD less than 4.0%, which accords with the level for bio-sample analysis.