ANALYTICAL BIOCHEMISTRY 123,402-407 (1982) Cord Blood Screening for Hemoglobin Disorders by High-Performance Liquid Chromatography*9’

Ion-exchange high-performance liquid chromatography was employed as a screening method for abnormal hemoglobins in the newborn period. Samples of cord blood collected in EDTA tubes were used for this analysis. Hemolysates were injected onto 4.1 X loo-mm Synchropak ion-exchange columns using an automatic injector. Hemoglobin separation was carried out by means of a sodium acetate gradient. A total of 415 samples was analyzed. Hemoglobins A, F, and Bart’s, as well as C or S when present, were separately eluted and quantitated using a 35min gradient program. Four individuals with sickle cell disease, 26 with S or C trait, one with SC disease, and two others with alpha-chain variants were diagnosed with this method. The proportion of Bart’s hemoglobin was greater than 1 % in 33 individuals. The elution pattern was highly reproducible. The potential for complete automation and the ease with which quality control can be assured make this technique well suited for the detection of abnormal hemoglobins in the newborn period. Neonatal screening for abnormal hemo-globins provides a practical approach for the detection of sickling syndromes and other hemoglobin disorders such as a-thalassemia. Most screening programs utilize various electrophoretic techniques for the detection of abnormal hemoglobins ( l-4). Electropho-resis on cellulose acetate at pH 8.6 identifies increased amounts of Bart’s hemoglobin (Hb) * associated with alpha thalassemia ( 1). However, hemoglobins F, A, and S are not always well resolved from each other. Elec-trophoresis on citrate agar at pH 6.2 pro-vides better separation of Hb F from Hb S; however, Bart’s Hb is not easily detectable. Some screening programs utilize both of these techniques because of the limitation