In this work, a real-time label-free microwave sensing mechanism for glucose concentration monitoring using a planar biosensor configured with an inset fed microstrip patch antenna has been demonstrated. A microstrip patch antenna with the resonating frequency of 1.45 GHz has been designed and is fabricated on the Flame Retardant (FR-4) substrate. Due to the intense electromagnetic field at the edges of the patch antenna, edge length has been used as the detecting area where the sample under test (SUT) interacts with the electromagnetic field. The Poly-Dimethyl-Siloxane (PDMS) with the trench in the centre has been employed as the sample holder. Here, the SUT is the glucose dissolved in DI (de-ionized) water with the concentration range of 0.2 to 0.6 g/mL. The dielectric constant dependency on the glucose concentration has been used as the distinguishing factor which results in a shift in the S-parameter. The experimentally measured RF parameters were observed closely which showed the shift in S11 magnitude from –40 to –15 dB and resonant frequency from 1.27 to 1.3 GHz w.r.t the SUT solution of 0.2 to 0.6 g/mL with linear regression coefficient of 0.881, and 0.983 respectively.