Detection of hemoglobinopathies by high-performance liquid chromatography in patients with microcytic hypochromic anemia in Western Nepal

Objectives: Hemoglobinopathies represent a significant but underdiagnosed cause of microcytic hypochromic anemia. High-performance liquid chromatography (HPLC) is the gold standard for detecting hemoglobin variants, yet routine screening remains limited in Nepal’s Terai region. This study aimed to detect hemoglobinopathies using HPLC and determine their prevalence at Nepalgunj Medical College. Material and Methods: A hospital-based cross-sectional study was conducted among 228 patients with microcytic hypochromic anemia (mean corpuscular volume MCV <80 fL) at a tertiary care hospital in Western Nepal from October 2023 to September 2024. Complete blood count was performed using and hemoglobin variant analysis was performed using Sysmex XN-1000 and Bio-Rad D-10 HPLC system. Internal and external quality assurance was followed according to manufacturer protocols. Hemoglobin fractions were quantified according to standard diagnostic criteria. Statistical analysis was performed using the Statistical Package for the Social Sciences version 26. Results: The mean age was 30.96 ± 17.28years with female predominance (75.9%). Mean hemoglobin was 8.21 ± 2.28 g/dL and MCV was 70.54 ± 7.36 fL. Among this selected hospital population with microcystic anemia, hemoglobinopathies were detected in 45 patients (19.7%). β-thalassemia trait was most common (23 cases, 10.1%), followed by sickle cell trait (17 cases, 7.5%), sickle cell disease (3 cases, 1.3%), and high fetal hemoglobin variants (2 cases, 0.9%). Mean hemoglobin A2 in β-thalassemia trait cases was significantly elevated (4.95 ± 0.66%) compared to non-thalassemia cases (2.38 ± 0.44%, P < 0.001). Conclusion: Hemoglobinopathies were detected in 19.7% of patients with microcytic hypochromic anemia. HPLC screening enables accurate diagnosis and facilitates genetic counseling, supporting its integration into routine clinical practice in high-prevalence regions.