We propose pulse power generation (PPG) amperometry as an advanced readout realized for Prussian blue (PB)-based (bio)sensors. In contrast to the conventional power generation mode, when the current response is generated upon continuous short-circuiting, the suggested pulse regime is fulfilled by periodic opening and shorting of the circuit. Despite PB being electroactive, the pulse readout is advantageous over conventional steady-state power generation, providing up to a 15-fold increased signal-to-background ratio as well as dramatically improved sensitivity exceeding 10 A·M–1·cm–2 for H2O2 sensors and 3.9 A·M–1·cm–2 for glucose biosensors. Such analytical performance characteristics are, most probably, achieved due to the enrichment of the diffusion layer by analyte mass transfer from the bulk upon opening of the circuit. Due to an improved sensitivity-to-background ratio, reduced flow-rate dependence, and enhanced operational stability, the regime allows reliable monitoring of blood glucose variations through sweat analysis with the on-skin device.