High Performance Liquid Chromatographic Separation of the Globin Chains of Non-Human Hemoglobins

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Abstract
High performance liquid chromatography has been applied to the separation of the globin chains of 16 non-human species which include common mammalian and avian species (Felis catus, Didelphis virginiana, Cavia porcellus, Elaphe obsoleta spiloides, Rana catesbeiana, Torpedo californica, Mus musculus, Rattus norvegicus, Mesocricetus auratus, Oryctolagus cuniculus, Capra hircus, Anas platyrhynchos, Columba livia, Melopsittacus undulatus, Canis familaris). The procedure uses a large-pore C4 column which has been effectively employed for the separation of human globin chains. In many cases, the gradient for human chains with trifluoroacetic acid-water-acetonitrile was satisfactory or required only moderate modification. The separations were excellent for all except dog hemoglobin. Many results substantiate prior published information about heterogeneity, amino acid composition, etc. of the individual hemoglobins. Additional data on some of these hemoglobins have also been obtained, and some previously unstudied hemoglobins have been examined.