Comparative studies of haemoglobin variants in normal, sickle cell trait, sickle cell disease and -thalassemia trait patients using cellulose acetate electrophoresis and cation exchange-high performance liquid chromatography

Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud University,

Riyadh 11451, Saudi Arabia

E-mail : manalkhalid2005@yahoo.com, sarjumand13@gmail.com, muskaanht@yahoo.com,

sfatima2013@yahoo.com, noura_jameil@yahoo.com

Manuscript received online 29 October 2013, revised 08 November 2013, accepted 09 January 2014

Approximately 5% of the world’s population carries trait genes for haemoglobin disorders, mainly, sickle cell disease and thalassemia. Haemoglobin disorders are genetic blood diseases due to inheritance of mutant haemoglobin genes from both, generally healthy parents. Sickle cell disease and β-thalassaemia are most common in eastern province of Saudi Arabia. The present study was undertaken to analyze the variation in levels of haemoglobin (Hb) types in different haemoglobin disorders in samples of patients from Riyadh, Saudi Arabia by CAE and HPLC method and to identify the best and effective method for diagnosis of different types of Hb disorders. In the present prospective study blood samples from four groups – normal healthy males and females, sickle cell trait, sickle cell disease and β-thalassemia patients were included. The levels of Hb variants such as HbA, HbA₂ , HbF and HbS were measured by two techniques – cellulose acetate electrophoresis (CAE) and cation exchange-high performance liquid chromatography (CE-HPLC). There was a significant increase (p < 0.001) in levels of HbA₂ in patients with β-thalassemia compared to normal and sickle cell trait/disease. The levels of HbF and HbS increased significantly (p < 0.001) in patients with sickle cell disease compared to other groups. The HPLC chromatogram of Hb variants acts as fingerprint for diagnosis of different haemoglobinopathies. The separation and analysis of different Hb fractions was achieved effectively using HPLC when compared to cellulose acetate electrophoresis. The present study facilitates clear understanding of changes in levels of Hb fractions in different types of haemoglobinopathies and also proves HPLC as the best technique that can be used for diagnosis of the disease. HPLC was found to be more efficient method due to high sensitivity and resolution compared to cellulose acetate electrophoresis.