Comparative Study between Hplc and Hb Electrophoresis to Characterize Hemoglobin Profile in Hemoglobinopathies.

Abstract

INTRODUCTION: Mutation or deletion of one of the globin genes of hemoglobin results in thalassemias and hemoglobinopathies. Two diagnostic modalities which can be used to study hemoglobin abnormalities is Hemoglobin electrophoresis and capillary zone electrophoresis. METHODS: A total of 41 adults and children blood samples were examined for routine thalassemia screening using both the modalities, excluding those patients who had undergone blood transfusion within a span of 12weeks. All samples were analysed using automated cell counter (Sysmex XN-1000), HPLC (Biorad D-10) and capillary zone electrophoresis. RESULTS: Out of 41 cases, 24 cases were of thalassemia trait, 9 cases were normal, 3 cases of β-thalassemia major, 2 cases of homozygous/ double heterozygous for β- thalassemia and δβ-thalassemia, 1 case of sickle cell anemia, 1 case of Compound heterozygous sickle cell anemia and β-thalassemia and 1 case of compound heterozygous for HbE/ β-thalassemia. When HbF value is >16.5% it merges with HbA1c values in HPLC(Bio-Rad D-10) because of which it is very difficult to comment about exact values of HbF with the help of HPLC. So for such cases CZE helped us to evaluate exact value of HbF. CONCLUSIONS: Forty one cases were collected in which both HPLC and Hb electrophoresis gave same results for 34 cases and remaining 7 cases we got different impressions from both the modalities. Advantage of Hb electrophoresis over HPLC is that it gives exact values of HbF which is very essential to differentiate thalassemia intermedia from thalassemia major and also it exactly evaluates percentage of HbE which elutes with HbA in case of HPLC. CZE is complimentary to HPLC, which is very helpful for evaluating exact values of HbF,HbS and HbE. DocuSign Envelope ID: FE404E2D4110C8D-4-C86749-94-344D78-6A-09A58E-FF-554091EE60CCFA5725A37CB