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

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 alpha, beta, and gamma chains. One of these had previously been identified as the delta chain. The material in the most prominent minor peak which is termed "pre-beta" is indistinguishable from the normal beta chain in amino acid composition, but the difference has not been identified. An analogous pre-delta peak appears when chromatographically isolated Hb A2 is examined, but pre-alpha and pre-gamma 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 gamma chains appear to do so.