In this work, a novel glucose quantification strategy is presented in a self-powered biosensing device. The analyte in the sample is oxidized in an enzymatic fuel cell and the generated charge is transferred to a capacitor. The built-up capacitor voltage at a specific time can be directly correlated with the concentration of analyte. An electro-fluidic switch placed on a paper-based microfluidic channel connects a minimalistic electronic circuit to the fuel cell. The circuit modules discriminate the built-up capacitor voltage, which corresponds to a particular glucose range. The digital semi-quantitative result is visualized vía electrochromic displays. As a practical application of this working principle, the self-powered single-use device has been designed to perform screening of gestational diabetes mellitus. The device discriminates between healthy (7.8 mm), and diabetes (>11.1 mm) condition providing a reliable result. This single use, printable, and disposable self-powered glucose biosensing device is autonomous and fully powered by the glucose contained in a 3.5 µL sample. It offers an energy-saving, environmentally friendly, and low-cost solution for Point-of-Care testing. By replacing the selective enzyme in the fuel cell, this strategy can be used for measuring other health parameters such as creatinine, cholesterol, or uric acid, among others.