Hydroquinone poses a major threat to human health and is refractory to degradation, so it is important to establish a convenient detection method. In this paper, we present a novel colorimetric method for the detection of hydroquinone based on a peroxidase-like Pd nanozyme. The vancomycin-stabilized palladium nanoparticles (Van-Pdn NPs, n = 0.5, 1, 2) were prepared using vancomycin as a biological template. The successful synthesis of Van-Pdn NPs (n = 0.5, 1, 2) was demonstrated by UV-vis spectrophotometry, transmission electron microscopy, and X-ray diffraction. The sizes of Pd nanoparticles inside Van-Pd0.5 NPs, Van-Pd1 NPs, and Van-Pd2 NPs were 2.6 ± 0.5 nm, 2.9 ± 0.6 nm, and 4.3 ± 0.5 nm, respectively. Furthermore, Van-Pd2 NPs exhibited excellent biocompatibility based on the MTT assay. More importantly, Van-Pd2 NPs had good peroxidase-like activity. A reliable hydroquinone detection method was established based on the peroxidase-like activity of Van-Pd2 NPs, and the detection limit was as low as 0.323 μM. Therefore, vancomycin improved the peroxidase-like activity and biocompatibility of Van-Pd2 NPs. Van-Pd2 NPs have good application prospects in the colorimetric detection of hydroquinone.