The main aim of our present study is to propose a surface plasmon resonance-based biosensor design for hemoglobin concentration monitoring in human blood samples. The proposed SPR sensor is based upon the Kretschmann configuration with silver metal, MXene, ZnO, and graphene nanolayers, with a BK7 coupling prism. A He-Ne laser optical input source with a 633 nm wavelength is used following the attenuated total reflection (ATR) principle. After employing the MXene, ZnO, and graphene layers over the conventional sensor setup, an investigation is done. With the optimized structure, our main aim is to detect the presence of hemoglobin concentration in the human blood with its refractive index variation. The calculated highest sensitivity of the proposed configuration is 154 \(Degree/RIU\) with the other parameters calculated as FWHM near \(3.69 Degree\), SNR of 0.5051 \({\text{D}\text{e}\text{g}\text{r}\text{e}\text{e}}^{-1}\) and quality factor of 41.58 \({RIU}^{-1}\).