In this approach, a reliable 2,6-dinitrophenol (2,6-DNP) sensor probe was developed by applying differential pulse voltammetry (DPV) using a glassy carbon electrode (GCE) decorated with a wet-chemically prepared PbO-doped ZnO microstructures’ (MSs) electro-catalyst. The nanomaterial characterizing tools such as FESEM, XPS, XRD, UV-vis., and FTIR were used for the synthesized PbO-doped ZnO MSs to evaluate in detail of their optical, structural, morphological, functional, and elemental properties. The peak currents obtained in DPV analysis of 2,6-DNP using PbO-doped ZnO MSs/GCE were plotted against the applied potential to result the calibration of 2,6-DNP sensor expressed by ip(µA) = 1.0171C(µM) + 22.312 (R2 = 0.9951; regression co-efficient). The sensitivity of the proposed 2,6-DNP sensor probe obtained from the slope of the calibration curve as well as dynamic range for 2,6-DNP detection were found as 32.1867 µAµM−1cm−2 and 3.23~16.67 µM, respectively. Besides this, the lower limit of 2,6-DNP detection was calculated by using signal/noise (S/N = 3) ratio and found as good lowest limit (2.95 ± 0.15 µM). As known from the perspective of environment and healthcare sectors, the existence of phenol and their derivatives are significantly carcinogenic and harmful which released from various industrial sources. Therefore, it is urgently required to detect by electrochemical method with doped nanostructure materials. The reproducibility as well as stability of the working electrode duration, response-time, and the analysis of real environmental-samples by applying the recovery method were measured, and found outstanding results in this investigation. A new electrochemical research approach is familiarized to the development of chemical sensor probe by using nanostructured materials as an electron sensing substrate for the environmental safety (ecological system).