HPLC Separation of Peptides of Human Blood

초록

Separation of biological-active substances is very important task in modern chemistry, biology, and medicine. Now there is information about the peptides, which has wide spectrum of biological activity. In the given work, we studied the chromatographic behavior of some peptides. These peptides were in the degradation process of human hemoglobin. Waters HPLC (U.S.A.) with UV-detector (l = 254 nm) was used. Two steel columns were Prosper 300 A / 5 mm C18 150x4.6 mm and Platinum EPS 100 A / 5 mm C18 150x4.6 mm. The extract peptides in trichloroacetic acid were used for the HPLC analysis. The separation of peptides was carried out in isocratic and linear-gradient mode of a mobile phase. The binary mobile phase was composed of water and acetonitrile. The mechanisms of chromatographic retention with various eluent were investigated. Influence of organic modifier on the retention mechanism, selectivity of chromatographic separation, pore-size effect in a sorbent were also carried out. The contributions of various variants of intermolecular interaction in chromatographic retention were calculated. We have chosen the optimal conditions for separation of peptides with the two chromatographic columns. The best gradients modes of mobile phase and types of eluent were chosen. The technique of chromatographic separation of peptides, which of human hemoglobin are generated, was offered. The identification of some biologically active peptides was made. Application of the HPLC for study of degradation process of human hemoglobin was shown.