Quantitation of hemoglobin components by high‐performance cation‐exchange liquid chromatography: Its use in diagnosis and in the assessment of cellular distribution of hemoglobin variants

Abstract

Additional data are presented that were obtained with a newly developed cation-exchange high-performance liquid chromatography (HPLC) method allowing the separation of numerous normal and abnormal human hemoglobin types. The method was found to be of considerable value for the diagnosis of certain hemoglobinopathies in the adult as well as in the newborn. Definitive differentiation between, AS, SS, S-beta+-thal, SD, SC, AC, C-beta+-thal, etc in cord bloods was readily accomplished even when the samples were collected on filter paper. Analyses of the hemoglobin in cells from Hb S and Hb C heterozygotes with different numbers of alpha chain genes (alpha alpha/alpha alpha; -alpha/alpha alpha; -alpha/-alpha) after separation by a Dextran density centrifugation technique failed to show significant changes in the relative quantitation of the variant hemoglobin types. Glycosylation of hemoglobin increased greatly with an increase in red cell age. Similar analyses of red cells from a young girl with Hb S trait and Hb H disease (the --/-alpha genic arrangement) gave comparable data. Some indication was obtained that her youngest cells contained some of the beta A4 and beta S4 tetrameric hemoglobins.