This research describes a novel electrochemical sensor based on a polyimide-silver nanowire (PI-AgNWs) composite that is used to modify a glassy carbon electrode (GCE) for the simultaneous detection of multiple nitrophenol isomers (NPs) with high performance. The synergistic effect from polyimide (PI) doped silver nanowires (AgNWs) improved the selectivity and sensitivity of the fabricated sensor. First, the AgNWs was prepared by polyol process, and then composited with PI to obtain the target composite. Subsequently, the composite was characterized in detail and was used to modify GCE for the NPs detection. Afterwards, electrochemical measurements were performed to simultaneous detection for three NPs. Under the optimal condition, the results exhibited that the linear detection range of 1-400 μM for 4-nitrophenol (4-NP), 1-350 μM for both 2-nitrophenol (2-NP) and 3-nitrophenol (3-NP) and the limit of detection (LOD) were 0.081 μM for 2-NP, 0.107 μM for 3-NP, and 0.063 μM for 4-NP, respectively. In addition, the fabricated electrode exhibited excellent reproducibility (RSDs of 2.85% for 2-NP, 2.71% for 3-NP, and 1.80% for 4-NP, respectively) and stability (RSDs of 6.17% for 2-NP, 2.36% for 3-NP, and 3.89% for 4-NP, respectively) over multiple measurements. Moreover, the sensor showed excellent anti-interference capability and was successfully applied to detect NPs analytes in real water samples with recoveries ranging from 91.1% to 109.0%. This study emphasizes the potential application of PI-AgNWs composites for the rapid detection of NPs in the environment.