Quantitation of hemoglobins Bart's, H, Portland-I, Portland-II and constant spring by anion-exchange high-performance liquid chromatography.

We introduce a new high-performance liquid chromatographic procedure that uses a specific anion-exchange column for the separation of hemoglobin (Hb) Bart's (gamma 4), Hb H (beta 4), Hb Portland-I (zeta 2 gamma 2), Hb Portland-II (zeta 2 beta 2), and the abnormal Hb Constant Spring (alpha 2 extended beta 2) in cord blood and adult red cell lysates. The method provides quantitative data for Hb Bart's in cord blood that correlate well with the alpha-globin gene status of the babies and can be used for an initial identification of alpha-thalassemic conditions. Quantitation of Hb Bart's from cord blood samples that are collected on filter paper and submitted as dried blood spots is unreliable. The separation of Hb H and Hb Bart's allows an evaluation of the synthesis of these two hemoglobin components in patients with Hb H disease.