High performance liquid chromatographic analysis of human hemoglobins and their polypeptide chains: its use in the identification of variants

A review is presented of two high performance liquid chromatography procedures useful in the study of human hemoglobin variants. One is a cation exchange procedure which allows the separation of normal and numerous variant hemoglobins as dimers, and the second a reversed phase method making it possible to separate polypeptide chains. The behavior of different abnormalities both as dimers with heme attached and as single polypeptides without heme at a low Ph value is reviewed. Numerous examples are presented to demonstrate the usefulness of both methods in studying hemoglobin abnormalities in a clinical setting; reviewed are subjects with α-thalassemia, β-thalassemia, with a combination of an α-chain variant and a β-chain variant, and with two β-chain variants. The two techniques have limitations, but can often provide useful information, particularly in conjunction with other methods.