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Herein, we fabricated a feasible and accurate sensing platform for the quantification of toxic organic pollutant 2-nitroaniline (2-NA) in water samples through electrocatalyst made up of bismuth molybdate (Bi2MoO6, BMO) functionalized carbon nanofiber (f-CNF) modified electrode. The preparation of BMO/f-CNF composite is of two methods, such as co-precipitation (C-BMO/f-CNF) and ultrasonication method (U-BMO/f-CNF). The physicochemical properties of the composites were characterized by XRD, FTIR, Raman, BET, FE-SEM, and HR-TEM techniques. At U-BMO/f-CNF, the charge transfer resistance was low (Rct = 12.47 Ω) compared to C-BMO/f-CNF because nanosized U-BMO particles correctly aim at the defective sites of the f-CNF surface wall. Further, the electrocatalytic activity of C&U-BMO/f-CNF composites was examined by cyclic voltammetry (CV) and differential pulse voltammetry techniques (DPV) for the electrochemical detection of 2-nitroaniline (2-NA). The U-BMO/f-CNF/GCE shows a higher cathodic current, wide dynamic linear range of 0.01–168.01 µM, and superior electrocatalytic activity with a low detection limit (0.0437 µM) and good sensitivity (0.6857 μA μM−1 cm−2). The excellent selectivity nature of U-BMO/f-CNF/GCE was observed in the presence of various organic pollutants and a few toxic metal cations. The practical applicability such as stability, repeatability towards 2-NA outcomes with accepted results. Besides, the practical viability of as proposed U-BMO/f-CNF sensor was investigated in soil and lake water samples delivers good recovery results. Hence from these analyses, we conclude that U-BMO/f-CNF/GCE potential for the determination of hazardous environmental pollutant 2-NA.