Hemoglobin D Punjab or HbD Los Angeles is one of the commoner hemoglobinopathies. In β-chain there is substitution at position 121 i.e.121 GluGln (Glutamic Acid is replaced by glutamine). The clinical spectrum of all the variants are usually ranges from asymptomatic state to mild hemolytic anemia and mild to moderate splenomegaly. Here, the retrospective study was carried out in general population of all the age groups in Dhiraj General Hospital, Vadodara, Gujarat. The duration of study was 8 months. Diagnosis of HbD Punjab was done by hemoglobin HPLC method with BIO-RAD D-10™ machine. Total 16 Hb D Punjab cases out of 1245 cases were found. Which were further categorized in Hemoglobin D trait (HbAD); Hemoglobin D disease (HbDD); Hemoglobin D-β–thalassemia (thalassemia trait/AD trait or DD homozygous) and Double heterozygous S and D (HbSD) entity. While HPLC reporting, it is very essential to keep these differentials of Hb D Punjab entity in suspected hemoglobinopathies because in today’s scenario there has been change in geographic distribution of population. A collective data from clinical history, complete blood count, HPLC finding & sickling solubility tests enable definitive identification of hemoglobin D Punjab variant.