In the quest for the development of highly sensitive and accurate optical biosensor for the detection of hemoglobin (Hb) concentration, theoretically we have investigated the effect of iii-v nitride on performance of graphene based SPR biosensor. To achieve the performance, computation has been performed for single metal and bi-metal individually for graphene based SPR configuration. Due to the addition of iii-v nitride layer in between metal and graphene layer in the proposed configuration, the overall sensitivity is enhanced and also detects very small volume of Hb. Finally, a comparative analysis has been carried out to establish that nearly 3:1 ratio bi-metallic combination (i.e. Ag38Au12) based SPR sensor can demonstrate better response. Additionally, introduction of indium nitride (InN) is more beneficial in comparison to other semiconductors. We expect that this study will motivate the researchers in the fabrication industry and helps physician to analyze Hb in real time. (C) 2016 Elsevier B.V. All rights reserved.