Introduction: Hemoglobinopathies are common genetic disorders of hemoglobin, which can be prevented by population screening and offering genetic counseling. The cumulative gene frequency of hemoglobinopathies in India is 4.2%. The carrier state for β-thalassemia in India varies from 1% to 17% with an average of 3.2%. The present study was undertaken to find out the burden of hemoglobinopathies and spectrum of this disorders among the blood donors. Materials and Methods: The study includes 975 students between 18 and 25 years who donated blood, were screened for β-thalassemia trait and related hemoglobinopathies by highperformance liquid chromatography (HPLC) using Bio-Rad variant. Samples were also run on a hemato analyzer for red cell indices and peripheral smear for red cell morphology. Results: A total of 41 donors showed abnormal hemoglobin fractions in HPLC. Out of these, 32 (3.3%) were diagnosed with β-thalassemia trait, 8 (0.8%) with Hb-D Punjab and 1 (0.1%) with Hb-S trait. The frequency of β-thalassemia trait in the student from different geographical regions varied from 0.8% to 4.44%, being the highest in those from Punjab and frequency of β-thalassemia trait in different caste groups varied from 0 to 4.74%, being the highest in the student from Jatt Sikh community. Conclusions: A universal approach of screening for β-thalassemia trait should be included as a part of standard blood testing among the college students, premarital and of the extended family of thalassemics. Population group with high gene frequencies requires screening programs as well as increased awareness and education program to control the birth of thalassemia major.