A microstrip highly sensitive differential sensor for complex permittivity characterization of urine samples was designed, fabricated and tested. The sensing area contains two pairs of open-stub resonators, and the working frequency of the unloaded sensor is 1.25 GHz. The sensor is easily implemented on an affordable substrate FR-4 Epoxy with a thickness of 1.6 mm. A Teflon beaker is mounted on the sensor without affecting the measurements. Numerically, liquid mixtures of water and urine at different percentages were introduced to the proposed sensor to evaluate the frequency variation. The percentage of water content in the mixture varied from 0% (100% urine) to 100% (0% urine) with a step of 3.226%, thus giving 32 data groups of the simulated results. Experimentally, the mixtures of: 0% urine (100% water), 20% urine (80% water), 33% urine (66% water), 50% urine (50% water), 66% urine (33% water), and 100% urine (0% water) were considered for validation. The complex permittivity of the considered samples was evaluated using a nonlinear least square curve fitting in MATLAB in order to realize a sensing sensitivity of about 3%.