Rapid procedure for globin chain analysis in blood samples of normal and beta-thalassemic fetuses

Abstract

Abstract Reversed-phase high pressure liquid chromatography (HPLC) was used for the separation of the alpha, beta, (A)gamma and (G)gamma chains from human blood samples. The alpha and beta chains were normally eluted close together, but their separation was improved by coupling 2 or 3 columns in series, or by increasing the temperature of the columns. This method has been applied for the determination of beta/gamma ratios in blood samples obtained at fetoscopy from normal pregnancies and fetuses at risk for beta-thalassemia. The values obtained by high pressure chromatography were similar but slightly lower than those found by carboxymethyl cellulose (CMC) chromatography. The average (G)gamma/(A)gamma ratio of the chains labeled after a 2-hr pulse with [3H] leucine was almost identical to the actual (G)gamma/(A)gamma measured by absorbance at 280 nm, indicating a constant rate of synthesis and accumulation of both globin chains in the first trimester fetus.