In this study, we demonstrated a one-pot synthesis of Cu nanoplates (Cu NPs) with high purity and ‘‘cleansurface”. Different from surfactant-assisted method, we tailored the architecture by controlling the ripeningtime of Cu seeds. Citrate is introduced as a structure-directing agent. Since the ‘‘clean surface” providemore available active sites, the as-prepared Cu NPs show a superior performance when utilized in thenon-enzymatic biosensors. In the detection of nitrite, the sensor constructed by Cu NPs exhibited a superiorsensitivity (303.1 ± 10.18 mAmM1cm2), and the detection range is 5.0 mM. On the contrary, sensitivityfor the Cu NPs by surfactant-assisted method is negligible. As applied in the glucose sensors,detection range by Cu NPs could reach 36.0 mM. And the sensitivity under ambient condition (298 K)is 561.93 ± 3.72 mAmM1cm2, which is ca. 2.11-folds higher than that for Cu NPs by surfactantassistedmethod. In the detection of H2O2, detection range is 10.0 mM, and the enhancement factor fordetection range is up to 2.0-folds. The superior performance can also be extended to the catalysis forthe reduction of 4-nitro-phenol. Since no additional noble metals was applied, this strategy would openup a new avenue for utilizing non-noble metals.