Health care workers (HCW) are a high-risk population for Hepatitis B infection. Hepatitis B vaccine which is 95% effective confers long-term protection and anti-HBs titre is a marker for protective immune response. Our objective was to assess the status of hepatitis B vaccination and to evaluate the anti-HBsAg titres among health care workers in a tertiary care hospital, Tamil Nadu. It was an observational study conducted among 610 Health care workers in a tertiary care teaching hospital from June to December 2018 after obtaining clearance from IEC. Workers were assessed for their HBV vaccination status and for their anti-HBsAg titre after getting informed consent. The antibody titres were measured using CLIA (chemiluminescent Immunoassay) supplied by Abbott diagnostics. The data was entered and analyzed using a Microsoft Excel sheet. In our study, 80.5% were fully vaccinated, 18.5% of them were defaulters which comprised the HCW with 2 doses and 1 dose of vaccine and 0.9% were not vaccinated. In the fully vaccinated group, 37% showed Anti HBs titres of 10-100 mIU/ml, 59.2% showed the titre of > 100 mIU/ml and 3.6% did not show the protective antibody titre (<10 mIU/ml ). The 3.6% who did not show protective antibody titre were given the booster dose of vaccine. Among those who received a booster dose, 61.1% responded with the titre of 10-100 mIU/ml and 22.2% responded with the titre of > 100 mIU/ ml and 16.6% did not respond even with the booster dose. In the defaulters 86.7% had titres < 10 mIU/ml , 9.7% had titres of 10 mIU/ml and 3.6% had titres > 10 mIU/ml. In the non vaccinated group all had titres < 10 mIU/ml. The present study emphasizes the importance of screening of anti-HBsAg titres to be made mandatory for all the health care workers along with HBV Vaccination.