Hydrophobic Interaction Chromatography in the Detection of Hemoglobinopathies

Abstract

We describe a rapid method of high-performance liquid chromatography that utilizes the principle of hydrophobic interaction to separate the mutant globin ß chains found in certain hemoglobinopathies. We used this method to analyse blood globin chains in patients with all the known diagnostized hemoglobinopathies found in Finland, that is Hb_Helsinki' Hb_Linköping' Hb_Vaasa' Hb_Hiiiyama and Hb_Syracuse. As reference we also analyzed normal adult and fetal blood hemoglobins. Our method allowed good separation of the mutant ß globin chains from intact chains in the heterozygous patients as well as from α globin chain and fetal γ globin chains. We used two different systems of liquid chromatographic columns for the analyses, each column giving a slightly different elution selectivity and complementing the other. The analyses we describe are rapid and automated and allow hemoglobinopathies to be diagnosed precisely and, owing to the different principles of separation, complement other methods in common use.