Environmental pollution has become a major human concern with the extensive exploitation of pesticides. Pentachlorophenol (PCP) is the most hazardous of all chlorophenols which are being used as pesticide, fungicide, and wood preservative. Thus, the fabrication of ultrasensitive electrochemical methods for the determination of pesticides is of great significance. In the present experiment, a simple, green, and sensitive electrochemical sensor was constructed for the determination of PCP by using a chemically modified nickel ferrite glassy carbon electrode (NiFe2O4/GCE). The fabricated nanoparticles were primarily characterized by several analytical tools to confirm the functionalities, surface texture, crystallinity, and elemental composition. For the investigation of conductive nature, the proposed NiFe2O4/GCE was exploited to the primary electrochemical characterization tools e.g. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The ultra-sensitive determination of PCP was carried out under the linear dynamic range from 0.01 to 90 μM at the pulse amplitude of 80 mV/s in BRB buffer pH of 4. The limit of detection of the developed methods for PCP was calculated to be 0.0016 μM. The analytical applicability of the fabricated sensor was tested in different water samples depicting the acceptable recovery values.