Analytical Determination of Hemoglobin by Spectrophotometric Method

Devolvement of rapid, simple, Sensitive, and stable spectrophotometric method for the assay of Hemoglobin using 2,4-dimethylaniline (DMA) and Hydrogen peroxide (H2O2) is presented. This study is based on the intermolecular coupling of oxidized DMA in the presence of H2O2 and Hemoglobin yields intense pink chromogenic coupled product having absorption maximum at 540 nm. The linearity of Hemoglobin by both rate and fixed time methods was found in the rage of 15.3 - 138 mM and 1.53 – 61.2 nM indicating the sensitivity of this method. H2O2 sensitivity range is found to be 20.2 - 331 μM, Michaelis-Menten constant for DMA and H2O2 were found to be 0.75 mM and 0.45 μM respectively. Catalytic efficiency and catalytic power were found to be .75 × 104 and 0.33 min-1 respectively. The reaction was applied for the assay of Hemoglobin and the results of proposed method correlated with Standard reference method.