Background: Nearly one million deaths reported globally are due to HBV (hepatitis B infection). Approximately more than 257 people have chronic hepatitis B infection across the globe which ultimately leads to hepatocellular carcinoma. Also, HBV infection is a major healthcare concern in developing nations such as India. Aim: The present study aimed to evaluate the prevalence of hepatitis B in voluntary blood donors at Indian healthcare centers. Methods: The present study assessed 10,000 blood donors who fit the inclusion criteria for the study and were accepted for blood donation within the defined study period. The blood samples collected from all the participants were evaluated for Hepatitis B surface antigen and other TTIs (transfusion-transmitted infections). Data gathered were analyzed statistically. Results: The results of the present study showed that among 10,000 donors assessed in the study, there was a prevalence of 0.62% for HBsAg-positive donors in all the subjects. HBsAg positivity rate was significantly higher in males with 0.7% compared to females and in the age range of 56-60 years with 1.5%. Conclusions: The present study concludes that the prevalence of hepatitis B is decreasing in voluntary blood donors in comparison to the previously reported data. It is vital to generate awareness and education concerning vigorous donor screening, voluntary blood donation encouragement, mass immunization, and blood-borne infections which might help in decreasing the load of hepatitis B and prevent further transfusion-transmitted infections.