High-pressure liquid chromatographic separation of glycosylated and acetylated minor hemoglobins in newborn infants and in patients with sickle cell disease.

High-pressure liquid chromatography on a synchropak CM-300 column was used for separation and quantitation of glycosylated and acetylated minor hemoglobins and major hemoglobins in 15 newborn infants, 17 patients with sickle cell anemia and one each with SC condition, CC disease, Hb S heterozygosity, and Hb C heterozygosity. Hemoglobin Bart's, Fla1, Fla2, Flb, Flc, FO, Ald, AO, and A2 have been separated from newborn infants. Similarly, hemoglobins Fla, Flb, Flc, FO, Sla+b, Slc, A2, Sld, and SO were separated from patients with SS condition. Minor hemoglobins in all the samples were also separated by conventional Bio-Rex 70 chromatographic methods. The percentages of the various hemoglobin components determined by high-pressure liquid chromatography agreed rather well with those obtained by Bio-Rex 70 chromatography. However, the quality of the resolutions and the sharpness of the peaks were, in general, superior with the high-pressure liquid chromatography approach, which is much faster, requires significantly less sample than Bio-Rex 70 chromatography, and is particularly suitable for fast analytical chromatography of minor and major hemoglobins.