Abstract Background Thalassemia and haemoglobinopathies pose a huge burden on Indian population, especially in Eastern India. Several studies indicated that abnormal hemoglobin variants influence Hb A1c estimation by High Performance Liquid Chromatography (HPLC) in patients with haemoglobinopathies. We opted capillary electrophoresis (CE) as an alternative method for Hb A1c measurement and aimed to evaluate Hb A1c test results done by (HPLC) with capillary electrophoresis (CE) to rule out the interference caused by abnormal hemoglobin variants. Methods A total of 3000 patients were tested for haemoglobinopathy in our laboratory from (1st -31th) January, 2025. Haemoglobinopathy were screened by Bio-Rad Variant II hemoglobin testing system. Both HPLC and CE methods were employed for Hb A1C testing in the group of patients (137/3000) with haemoglobinopathy using Bio-Rad Variant II Turbo and Sebia Minicap Flex Piercing respectively. Reference range defined for Hb A1c by HPLC: Non-diabetic: 4-5.7, Pre-Diabetic:5.7-6.4, Diabetic =6.5 and reference range for HbA1C by CE: 6 %. Statistical interpretation by was done by SPSS version 26.0. P value less than 0.05 was considered as significant. Statistical correlation was obtained by using Linear regression study. Results : 4.5% (137/3000) of total patients were reported to have abnormal hemoglobin variants in their HPLC chromatogram. 52% (72/137) Hb E carrier, 7.2% (10/137) homozygous E, 23.3% (32/137) beta thalassemia carriers, 1.4% (2/137) beta thalassemia major, 7.2% (10/137) Hb S carrier, 1.4% (2/137) homozygous S ,5.1% (7/137) Hb D carrier and 1.4% (2/137) HPFH (hereditary persistence of fetal hemoglobin) carrier comprised the group of haemoglobinopathies. We compared Hb A1c test results measured by HPLC and CE techniques. Hb A1c test values measured by using both the techniques were observed to be within reference range (Mean±SD: 5.24±1.05 by HPLC, Mean±SD: 5.19±1.08 by CE) in all patients with haemoglobinopathy. Hb A1c tests conducted in patients with haemoglobinopathies by HPLC and CE methods showed strong correlation in test results (Hb E trait: r=0.75 at p =0.01; beta thalassaemia traits: r=0.89 at p=0.04; Hb D: r=0.94 at p=0.2 Hb S: r=0.78 at p=0.21). In contrast to many reported studies, no observable difference was found in Hb A1c values in persons with haemoglobinopathies. HPLC and CE, both the methodologies failed to detect Hb A1c values in homozygous E, homozygous S and beta thalassaemia major patients. The study is limited by less number (n) of participants in each category which might attribute to high p value for determining level of significance. Conclusion Hb A1c test results done by HPLC and CE were in perfect concordance with each other. No such interference by abnormal haemoglobin variants on Hb A1c test results was observed in our study using HPLC method. Replacement of HPLC by CE as an alternative methodology is not necessitated rather both the methods can interchangeably be used for Hb A1c testing in patients with haemoglobinopathies. However, the present study is limited with small number of patients. An extensive study with larger participants is planned to validate the results claimed by our pilot study.
Rinini Dastidar (Wed,) studied this question.
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