Chemically prepared PrAlO3 perovskite nanoparticles (NPs) were applied for the electrochemical detection of warfarin, which is commonly utilized for preventing blood clots, such as in deep vein thrombosis. PrAlO3 perovskite NPs were synthesized by the co-precipitation process at environmental conditions. Crystallographic structure, phase purity, morphological structure, thermal stability, optical properties, and electrochemical characteristics were investigated by X-ray diffraction (XRD), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), Fourier transform infrared (FTIR) spectroscopy, UV-visible analysis, and cyclic voltammetry techniques. TEM micrographs showed the highly crystalline structure, smooth surface, irregular shape, and size of nanocrystalline particles with an average size of 20–30 nm. Particularly crystalline perovskite NPs were pasted on glassy carbon electrodes (GCE) to electrochemically detect the warfarin contents in liquid samples. The fabricated electrode was electrochemically characterized by different parameters such as different potential, scan rates, same potential with seven consecutive cycles, time response, real-time sample analysis, and as a function of warfarin concentration in phosphate buffer solution (0.1 M PBS, pH 7.2). The electrochemical electrode was further verified with various potentials of 5, 10, 20, 50, 100, and 150 mV/s, which exhibited sequential enhancements in the potential range. For detecting warfarin over a wide concentration range (19.5 µM–5000 µM), the detection devices offered good sensitivity and a low limit of detection (19.5 µM). The time-dependent influence was examined using chronoamperometry (perovskite NPs/GCE) in the absence and presence of warfarin at four distinct voltages of +0.05 to +1.2 V from 0 to 1000 s. The repeatability and reliability of the constructed electrochemical sensing electrode were also evaluated in terms of cyclic response for 30 days, demonstrating that it is substantially more reliable for a longer period. The fabricated perovskite NPs/GCE electrodes could be employed for the rapid identification of other drugs.