Uncovering Silent Hemoglobinopathies: Utility of HPLC in Outpatient Screening

Hemoglobinopathies are among the most common inherited red cell disorders globally and pose significant publichealth burden in India. Differentiating hemoglobinopathies from nutritional anemia, particularly in antenatal women, remains adiagnostic challenge. The aim was to determine prevalence and spectrum of undiagnosed hemoglobinopathies among patientsattending outpatient clinics using high-performance liquid chromatography (HPLC). Methods: A cross-sectional observationalstudy was conducted from January 2022 to December 2024. A total of 7,440 blood samples from patients attending generalmedicine, pediatrics, and antenatal OPDs were analyzed. Patients with a known diagnosis of hemoglobinopathy, recent bloodtransfusion (within 1 year) and patients with uncontrolled diabetes were excluded. Complete blood count (CBC), peripheralsmear, sickling tests, and HPLC chromatograms were analyzed. Results: A total of 7,440 outpatient samples were analyzed anda female preponderance (81.2%) was seen in the study population. Among these, 162 cases (2.17%) were found to have abnormalhemoglobin (Hb) fractions suggestive of hemoglobinopathy on HPLC. The most common hemoglobinopathy identified was theheterozygous beta-thalassemia trait (1.37%), followed by heterozygous HbAE (0.32%), heterozygous HbAS or sickle cell trait(0.18%), and heterozygous HbAD Punjab (0.14%). Rare Hb variants included homozygous sickle cell disease (HbSS) (0.04%),hereditary persistence of fetal hemoglobin (HPFH) (0.05%), and 1 case each (0.01%) of homozygous HbEE, heterozygous HbAC,compound HbE/beta-thalassemia, and delta–beta thalassemia. Additionally, 5 cases (0.06%) showed unidentified abnormalpeaks on HPLC that required further molecular testing. Peaks in the P03 and P04 retention windows were correlated withelevated HbA1c in individuals with diabetes and excluded from hemoglobinopathy classification. Conclusion: HPLC is areliable, rapid, and sensitive tool for detecting both common and rare hemoglobinopathies in outpatient settings. Its integrationinto routine antenatal and general screening can facilitate early diagnosis, genetic counseling, and preventive interventions.Expanding access to HPLC testing can reduce the burden of transfusion-dependent hemoglobinopathies and improve long-term patient outcomes.