A study of the minor peaks in high performance liquid chromatograms of globin chains on reversed phase columns

Abstract

The high performance liquid chromatograms of the hemoglobins in a hemolysate show minor peaks on reversed phase columns in addition to the expected major peaks of the α, β, and γ chains. One of these had previously been identified as the δ chain. The material in the most prominent minor peak which is termed “pre-β” is indistinguishable from the normal β chain in amino acid composition, but the difference has not been identified. An analogous pre-δ peak appears when chromatographically isolated Hb A₂ is examined, but pre-α and pre-γ peaks have not been observed. One minor peak contains a non-heme protein(s). Glycosylated hemoglobins do not separate from their non-glycosylated counterparts, but acetylated γ chains appear to do so.