Introduction: Thalassemia and other hemoglobinopathies are the most common routinely encountered congenital anemias in India. Sickle cell anemia is very common, so a study to rule out all the hemoglobinopathies is essential. Aims and Objectives: The present study was conducted to diagnose various hemoglobinopathies among clinically suspected cases., the present study was conducted to diagnose various hemoglobinopathies among clinically suspected cases. Materials and Methods: A total of 212 cases cases were undertaken in this study after they met the inclusion and exclusion criteria. HPLC was performed on the samples received in the Central Clinical Laboratory of Dr. Vasantrao Pawar Medical College, Hospital and Research Centre, Nashik, Maharashtra. Results: Out of 212 cases, 42 cases showed positivity for thalassemia or presence of abnormal hemoglobin, with 8% cases of Beta thalassemia trait, 1.88% cases of Beta thalassemia major, 4.7% cases of Sickle cell trait, 4.24% cases of Sickle cell disease and 0.94% cases of Sickle-thalassemia. Conclusions: HPLC is a fast, reliable, and cost-effective method to diagnose any suspected case of hemoglobinopathy.