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

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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 .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.