β-Agonists fed to animals for human consumption pose a serious threat to human health. Fast, broad-spectrum detection methods are needed for on-site screening of various types of β-agonists from animal feeds, meats, and animal body fluids. We developed a colorimetric assay that uses gold nanoparticle (AuNP) plasmon absorption to realize quick detection of β-agonists from liquid samples. β-Agonists showed the capability of directly reducing HAuCl(4) into atomic gold, which involved oxidation of the amine or phenol group on the benzene ring of the β-agonists. The resulting atomic gold formed AuNPs spontaneously, which had strong plasmon absorption at 528 nm. The linear relationship between the concentrations of β-agonists and the AuNPs plasmon absorbance granted quantitative determination of β-agonists in solution. The AuNPs colorimetric assay showed different sensitivities toward β-agonists with different substituent groups on the aromatic ring. β-Agonists with phenol groups had a lower limit of quantitation (LOQ) than those with amine groups. High-resolution transmission electron microscopy (TEM) images revealed the sizes of the AuNPs were in the range 15-25 nm, while X-ray energy-dispersive spectroscopic data suggested the smaller particles observed in TEM with lower contrast may be salt particles from the buffer solution. The developed colorimetric assay can potentially be used for the detection of β-agonists and their analogues from serum, urine, and other liquid samples in the presence of interference from common antibiotics and glucose.