Research on Proteolytic Degradation of Hemoglobin of Human Blood by RP-HPLC

초록

The information of hemoglobin by a source of the biological peptides has appeared recently. Researches on this peptides become interesting and urgent. Now, in the literature there are some data about the degradation process of hemoglobin, which can form biological active peptides called enzyme. However, the formation of such peptides in human blood has been investigated very insufficiently. In the given work, we studied the formation of some peptides, which are the fragments of human hemoglobin with RP-HPLC. Blood of the healthy man was used as a sample material. Biological material was made with application of a technique, which was described in work [1]. The extracts of peptides were included in trichloroacetic acid. Chromatographic experiments on Waters HPLC (U.S.A.) with UV-detector were performed. The variant of reversed-phase liquid chromatography was utilized. With use of two chromatographic columns of Prosper 300A/5 mm C18 150x4.6 mm and Platinum EPS 100A/5 mm C18 150x4.6 mm, the analysis was made. Separation of peptides was carried out in linear-gradient mode of acetonitrile. The flow rate of mobile phase was 1 ml/min. UV detection of peptides was made at 254 nm. It was found that the cell of human erythrocyte can secrete short peptides. These peptides were formed because the degradation process of hemoglobin was carried out. This process is dynamic and of energy-dependence. It demonstrated that in the preserving process of human blood, the quantity of the peptides kept changing. The identification of some biologically active peptides was made.