Degradation of the Proteolytic Hemoglobin by High Performance Liquid Chromatography

초록

In this work, the hemoglobin degradation analyzed in non-cell model system was researched with RP-HPLC. The analytical HPLC system in these experiments was an Waters 600S Controller equipped with dual wavelength absorbance UV detector. The data acquisition system was Millenium32. The variant of reversed-phase liquid chromatography was utilized. The wavelength was fixed at 254 nm and the experiment was performed at room temperature. The size of the analytical column packed by C18 was 150×4.6mm (5㎛) (Althech Co., USA). The gradient mode was employed to isolate peptides. The flow rates of the mobile phase were fixed at 1 ml/min. The constant injection volume, 10 l, was used throughout the experiments. It was found that the cell of the human erythrocyte could secrete the short peptides. These peptides were formed because the degradation process of hemoglobin was carried out. The study of the low-molecular peptide excretion process was made by incubation of the erythrocyte cell in energy-dependence condition. This process is dynamic and of energy-dependence. It demonstrated that in the preserving process of the human blood, the quantity of the peptides kept changing. The opportunity and probability for the formation of the short peptides inside the human erythrocytes can be appreciated by the analysis of the peptides in non-cell model system (in vitro). The five peptides in the extracts were identified. The kinetics of the formation of the peptides was also investigated by RP-HPLC. These processes were described by the equation for first-order reaction. The obtained results lead that the formation and accumulation of the peptides in human blood occurs during the excretion processes of the peptides. We can chow, that the formation of the some peptides has the specific character.