In this study, a nanostructured CuSn(OH) 6 microsphere decorated on reduced graphene oxide (CSOH/rGO) was prepared by a co-precipitation method followed by ultrasonication, which was used for the sensitive and selective determination of L-Tryptophan (L-Trp) in the presence of uric acid. Its structure and morphology were characterized by X-ray diffraction (XRD), field emission scanning/transmission electron microscopy (FE-SEM/ TEM), Fourier transform infrared (FT-IR) spectroscopy, Brunauer−Emmett−Teller (BET) analysis, and X-ray photoelectron spectroscopy (XPS). Galvanostatic charge/discharge tests, cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV) methods have also been employed to investigate the electrochemical performance of the CSOH/rGO composite. Analytical parameters such as loading amount of catalyst, scan rate, pH value, cyclic stability, repeatability, and reproducibility were optimized. The CSOH/rGO composite shows excellent detection of L-Trp in a wide dynamic linear range (WLR) of 0.05−175.8 μM, a low limit of detection (LOD) of 2.0 nM, a limit of quantification (LOQ) of 68.9 nM, and a sensitivity of 0.21 μA μM −1 cm −2 . The fabricated electrode was further used to monitor the selectivity of L-Trp in the presence of uric acid and applied to the detection of L-Trp in spiked real samples with satisfactory results.