Background: As the life expectancy in thalassemia is improving, pain is being recognized as an emerging problem. There is a need for a prospective observational study of pain in these high-risk patients. Objectives: Assess the pain prevalence, severity, and impact of co-morbidities in thalassemia. Methods: All patients >10 years of age (n=165) attending Thalassemia Day Care Center of a tertiary care hospital were assessed for pain prevalence, severity, and its impact on various life activities using Brief Pain Inventory. Their medical records were reviewed for the presence of various comorbidities. All statistical calculations used SPSS 21 version statistical program for Microsoft windows. Results: The pain was reported by 62.4% of participants with 35.2% and 59.4% participants, reporting pain in the past 1 week and 4 weeks respectively. A significantly higher pain prevalence was reported in females (p=0.03), patients residing in urban areas and employed participants (p=0.03). The commonest sites of pain were the lower back and calves. General activity (p=0.02) and enjoyment of life (p= 0.02) were significantly affected due to pain in patients between 21 to 30 years of age. Female participants reported interference of pain with mood (p=0.03). A significant relation of pain prevalence was found with higher average serum ferritin (p = 0.015), moderate to severe liver iron concentration (p= 0.04), and lower levels of 25 hydroxyvitamin D levels (p=0.03). Conclusion: Pain is an emerging cause of morbidity in thalassemia. The study found a significant association of pain with modifiable factors such as serum ferritin, LIC, 25 hydroxyvitamin D levels.