Nitrogen doped porous carbon nanopolyhedra (N-PCNPs) were prepared from direct carbonization of ZIF-8 nanopolyhedra. The N-PCNPs showed uniform morphology, narrow pore-size distribution centered at 3.7 nm, high surface area (2221 m² g⁻¹) and good electrochemical properties and were used to modify a glassy carbon electrode to electrochemically detect ascorbic acid (AA), dopamine (DA) and uric acid (UA). Compared with the bare GC electrode and reduced graphene oxide modified glassy carbon (rGO/GC) electrode, the N-PCNPs modified GC electrode (N-PCNPs/GC) was found to perform better toward electrocatalytic oxidation of AA, DA and UA. For simultaneous sensing of three analytes, three well-separated voltammetry peaks were obtained using the N-PCNPs/GC electrode in differential pulse voltammetry measurements, and the corresponding peak separations between AA and DA, DA and UA were 228 mV, 124 mV respectively. The linear response ranges for the determination of AA, DA and UA were 80–2000 μM, 0.5–30 μM and 4–50 μM, respectively, and the detection limits (S/N = 3) were 740 nM, 11 nM and 21 nM, respectively. Furthermore, the N-PCNPs/GC electrode showed good reproducibility and stability. The attractive features of N-PCNPs provided potential applications in the simultaneous determination of UA, AA and DA.