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.