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[Display omitted] • Highly effective hybrid Ag@Pani/O-CMC-nanocomposite-based electrochemical sensor. • Impregnation of Ag@Pani to O-CMC matrix promotes sensing performance. • Trace detection of 4-Nitrophenol with LOD of 0.58 nM. • Remarkably sensitivity of 2.48 µA nM−1 cm−2 with high stability and great selectivity. Present work aims at fabricating chemically active, reproducible, and reliable electrode material by Ag nanoparticles anchored oxidized-carboxymethyl cellulose (O-CMC) embedded Pani (Ag@Pani/O-CMC) based nanocomposite for the trace detection of 4-nitrophenol. The Ag@Pani/O-CMC was physiochemically characterized in detail using FTIR, SEM, TEM, UV–vis, XPS, and XRD to analyze functional groups, structural morphology, optical properties, and chemical compositions. Differential pulse voltammetry and cyclic voltammetry were adopted to explore the electrochemical performance of Ag@Pani/O-CMC modified glassy carbon electrode (Ag@Pani/O-CMC/GCE). The observed reduction peak current was proportional to the 4-NP concentrations with a high sensitivity (2.48 µA nM−1 cm−2) and displayed a detection limit of 0.58 nM and a good linear range of 10–100 nM. In addition, the sensor exhibited comparable sensing capability in real biological samples, including milk, mixed fruit juice, and tap water, with excellent recovery from 97.20 % to 101 %. It also depicted high repeatability as the current response remained at 84.52 % after 28 days of operation for 4-NP detection. This approach can be projected to result in the electrochemical sensor for safe industrial processing and environmental monitoring, which is expected to have good stability, excellent selectivity, and sensitivity for detecting traces of 4-NP in actual samples. [ABSTRACT FROM AUTHOR]