Introduction: Hepatitis B is the world’s prevalent blood-borne viral infection, specifically it is prevalent with its two flip sides of the coin such as in health care workers and pregnant women. Since health care workers have a vital role in providing health care, thus are more vulnerable to occupational risk. On the flip side, the most common method of transmission of HBV around the world is from mother to infant. Hence, both population knowledge and practice should be at an optimal level. The purpose of this study was to assess the influence of the system side knowledge and practice towards Hepatitis B be on the demand-side population in Sabarkantha, Gujarat. Materials and Methods: This cross-sectional study was carried out in the Sabarkantha district of Gujarat. Data were obtained by using a piloted structured questionnaire in vernacular language from the supply-side (health workers) and demand-side (antenatal/ postnatal mothers) to assess the Knowledge of Hepatitis B at a point in time. Results: Study found 6.14 (± 2.89) and 1.69 (± 1.7) mean knowledge score (total score=14) towards the Hepatitis B disease, in the supply-side population and in the demand-side population respectively. The difference was statistically significant. In both populations, the most usual knowledge for symptoms and transmission of hepatitis reported were for Jaundice and hepatitis vaccine. Though the 60.9 % ANC/PNC revealed ASHA, as the most common source of information, the influence of ASHA’s knowledge on ANC/PNC was not up to the mark. Furthermore, the study noticed ambiguity between Hepatitis A, E and Hepatitis B knowledge in both populations. Conclusion: There is need of frequent context tailored training of Hepatitis B amongst supply-side workers.