The present work reports a sensitive and simple sensor based on an electrochemically treated pencil graphite electrode (pre‐PGE) modified with multi‐walled carbon nanotubes (MWCNTs) for electrochemical determination of L‐tryptophan (L‐Try) detection in the presence of uric acid. Electrochemical and physical characterizations of the MWCNTs/pre‐PGE were investigated using cyclic voltammetry (CV), Electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM) techniques. Under the optimized conditions, the linear working range was 0.030–200.0 μM with a limit of detection of 9.0 nM using the differential pulse adsorptive stripping voltammetry (DPAdSV) method. The proposed sensor exhibited good sensitivity, repeatability, reproducibility, and stability. The MWCNTs/pre‐PGE were successfully applied to L‐Try determination in commercial poly‐amino acid supplements with good recovery and accuracy.