Diagnostic Utility of High-Performance Liquid Chromatography and Its Correlation With Hematological Indices in the Evaluation of Hemoglobinopathies

Introduction: Hemoglobinopathies are common genetic disorders in India and frequently present as microcytic hypochromic anemia, often mimicking iron deficiency anemia. High-performance liquid chromatography (HPLC) enables accurate quantification of hemoglobin fractions and facilitates reliable diagnosis. Methodology: This cross-sectional study included 150 patients with microcytic hypochromic anemia (mean corpuscular volume (MCV) < 80 fL, mean corpuscular hemoglobin (MCH) < 27 pg) and clinical suspicion of hemoglobinopathy. Complete blood count and HPLC (Bio-Rad D10, Bio-Rad Laboratories, Hercules, CA) were performed. Hemoglobin fractions were quantified, and hematological indices were correlated with HPLC findings. Statistical analysis was performed using independent t-tests and chi-square tests. Results: Abnormal hemoglobin patterns were identified in 100 (66.7%) patients. Beta thalassemia trait was the most common finding, seen in 63 patients (42.0%), and was characterized by elevated HbA2 levels (mean: 5.2% ± 0.8%). These patients showed significantly lower MCV and MCH values and higher red blood cell (RBC) counts compared with those with normal HPLC patterns (P < 0.001). Beta-thalassemia major was identified in 12 (8.0%) with markedly elevated fetal hemoglobin (HbF) (mean: 85.6% ± 8.2%). Structural variants, including HbS trait (7, 4.7%) and homozygous HbE (6, 4.0%), were also detected based on characteristic HPLC patterns. Conclusions: HPLC is a reliable and precise diagnostic tool for hemoglobinopathies, with a strong correlation to hematological indices. Its use enables accurate differentiation of thalassemia carriers from other causes of microcytic anemia, supporting its role in routine diagnostic evaluation.