Introduction: Hepatitis B infection is the most transmissible blood borne infection and also a vaccine preventable disease. Health care worker (HCWs) are at increasing risk of acquiring Hepatitis B infection (10-30%). This study aims to estimate the awareness regarding various aspects of hepatitis B vaccination such as vaccination status, protective antibody titre and the prevalence of hepatitis B infection). Materials and method: Study included 400 HCWs (250 doctors, 100 nurses, and 50 technicians) of Lady Hardinge Medical College & Associated Hospitals, New Delhi. A self structured pretested questionnaire was used as a tool in the study. The level of awareness was graded (adequate, fairly adequate and inadequate) on the basis of their knowledge. Blood samples were collected from all the participants & subjected to ELISA test for detection of Hepatitis B surface antigen (HbsAg) and estimation of anti Hepatitis B antibody titer (anti HBs Ab) titre. Results: 34.4% doctors, 18% nurses and 4% technicians had overall adequate awareness level regarding various aspects of Hepatitis B vaccination. The difference between adequate awareness level among all the three groups of HCWs were significant (P