Quantitative microchromatographic determination of hemoglobin f in patients with hemoglobins s and/or c

Abstract

The improved microchromatographic procedure for the detection of Hb‐S and/or Hb‐C in cord blood at birth (Schroeder et al.: J. Lab Clin Med 86:528‐532, 1975) as well as a modification thereof may also be used for the quantitative determination of Hb‐F in the presence of Hb‐S and/or Hb‐C. However, Hb‐A interferes and must be absent. The methods use 0.5 × 6 cm columns of CM‐cellulose with Tris or Bis‐tris developers and require 2‐4 hr to complete. At low percentages of Hb‐F, the sharper zone of the Tris method is more easily visible than that of the Bis‐tris method, but the latter is a somewhat more rapid procedure. About 300 cases with Hb‐S and/or Hb‐C have been examined by the micro‐ chromatographic procedure. Most of these results (F_micro) have been compared with data from the determination of Hb‐F by one or more of the following methods: alkali denaturation (F_AD), conventional DEAE‐Sephadex chromatography (F_DES), or isoleucine analyses of zones from DEAE‐Sephadex chromatography (F_Ile). The accuracy and precision of the microchromatographic method is estimated to be 5‐10%. The microchromatographic methods require much less time than conventional chromatography but more time than alkali denaturation procedures. Compared to the latter, the new methods use whole blood and less blood and permit the physical separation of Hb‐F.