Liquid Chromatographic Assay of Plasma Hemoglobin Based on Oxidative Quenching of Serotonin by Pseudoperoxidase Effect of Hemoglobin

Abstract

A high-performance liquid chromatographic method for the determination of hemoglobin is described. The method uses the pseudoperoxidase effect of hemoglobin in the presence of hydrogen peroxide and is based on the oxidative quenching of the native fluorescence of serotonin. The determination range for hemoglobin was between 3 and 100 mg/dl. The intra-assay coefficient of variation was 2.04% or less, with a mean recovery of 102.0% at hemoglobin concentrations of 17.28, 43.20, and 69.12 mg/dl. A close correlation was found between the results of this method and the leuco crystal violet method.