Separation of adult chains of abnormal haemoglobin: Identification by reversed-phase high-performance liquid chromatography.

Rare abnormal haemoglobin (Hb) often poses a problem in the diagnosis of relative electrophoresis mobility. The rare neutral Hbs interact with the Hb S and thus can increase the severity of sickle cell anaemia. In the present study, we investigated the use of reversed-phase high-performance liquid chromatography (RP-HPLC) as a method for the identification of neutral or unstable Hb, and compared it with the well-known techniques urea-triton electrophoresis (UT) and isoelectrofocusing (IEF). The results of our experiments showed that Bucuresti Hb was separated without ambiguity by RP-HPLC. To date, no other electrophoresis techniques have been able to isolate this Hb. We also examined the Hb Nouakchott, which was the most hydrophobic Hb. Moreover, the adult and foetal chains of globins (especially the Agamma(t) and Agamma(i) chains) were quantified on the same chromatogram. Copyright 2004 Wiley-Liss, Inc.