Abstract This study presents a new method for detecting the organophosphorus pesticide glyphosate using advanced scanning electrodes (SPE) and enhanced fluorescence. Metal-organic frameworks from cobalt ions were synthesized using a solvothermal method. It is characterized using Raman spectroscopy, FT-IR, and X-ray diffraction techniques. The electrocatalytic behavior of the materials was studied using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Differential pulse voltammetry (DPV) examined the positive response of plants to glyphosate over a concentration range of 0.55–5.95 mM with a detection limit of 0.334 mM. The fluorescence enhancement ranges from 0.07 mM to 0.67 mM, and the detection limit is 0.0998 mM. Additionally, the selectivity of the proposed opto-electrochemical sensor was evaluated. This selection demonstrates the sensor's ability to detect glyphosate in complex wastewater matrices. This has important implications for environmental monitoring. By addressing glyphosate contamination, the sensor could significantly advance ecological remediation and monitoring strategies. The selectivity, sensitivity, and ability to operate under harsh conditions represent a significant advance in the development of efficient and reliable glyphosate technology for wastewater treatment and environmental protection. In real-sample matrices, the suggested sensor showed a good recovery of the pesticide that had been spiked.