Detection of haemoglobin variants for the diagnosis of beta thalassaemia & other haemoglobinopathies using anion exchange high performance liquid chromatography.

We describe the precise identification, separation and quantitation of several haemoglobin (Hb) variants such as Hbs A2, A, F, S, E, J, H, D-Punjab and Q-India in untransfused patients with beta thalassaemia and other haemoglobinopathies prevalent in the Indian population using whole blood (5 microliters), red cell lysates (2 microliters) or dried blood specimens on filter paper, using a conventional anion exchange high performance liquid chromatography (AX-HPLC) system. Samples were simultaneously studied for the presence of haemoglobin variants by alkaline electrophoresis. The problems in analysing rare variants and separating the closely migrating variants, encountered in the conventional methods could be resolved using the HPLC technique.