Broadband differences interferometeric analysis of a three-layer planar polymer optical waveguide is proposed and optimized to detect the concentration of hemoglobin in blood. The dispersion characteristic and cutoff film thickness of proposed waveguide are obtained by matching the field at various boundaries. The obtained cutoff film thickness for TE 0 and TM 0 modes is 0.09 µm, 0.1 µm at operating wavelength 400 nm, and 0.19 µm and 0.23 µm at operating wavelength 800 nm, respectively. The effective refractive indices of TE 0 and TM 0 modes are obtained at two considered wavelengths, i.e., 400 nm and 800 nm, and hence the difference of their propagation constant is calculated. It is observed that the propagation constant of these modes decreases with the increase of wavelength. Also, the difference of propagation constant attains its maximum value at certain wavelength and decreases either side of this wavelength. The interference maxima signals at output are considered as sensing signal. The maxima of interference signals, close to the maximum value of propagation constant, are shifted sufficiently with the change in cover refractive index. The maximum sensitivity 3.8 nm/RIU is obtained in the proposed broadband differences interferometeric analysis of waveguide at film thickness 300 nm. Hence, at this film thickness the sensing signal changes by 0.68 nm/g/L of hemoglobin concentration in blood.