An electrochemical sensor based on hierarchically Cu-BTC MOF material for non-electroactive glyphosate detection has been firstly constructed in this report. By using Cu-BTC frameworks as a detection matrix, the large specific surface area of the material can increase the electrode reaction site and further improve the detection performance. The electrochemical behavior of Cu-BTC frameworks for glyphosate detection was evaluated by cyclic voltammetry (CV). Under optimum conditions, differential pulse stripping voltammetry (DPV) is employed to detect glyphosate. The results show that the fabricated sensor displays ultralow detection limit (1.4 × 10−13 mol L-1) and wide detection range (1.0 × 10-12 to 1.0 × 10-9 mol L-1 and 1.0 × 10-9 to 1.0 × 10-5 mol L-1). Besides, this sensor possesses acceptable reproducibility and stability, as well as good selectivity against the major metabolite of glyphosate aminomethylphosphonic acid (AMPA) and other interference. Furthermore, this electrochemical sensor can also be applied to the detection of glyphosate in soybean. It is worth mentioning that this effectively Cu-BTC based sensor has great potential application in favorable and selective detection of organophosphorus pesticides in actual samples.