Demands for an evidence-based healthcare increase and today all medical decisions are to be based on scientific results. The evidence-based healthcare means that hospital librarians have a stronger role as mediators of scientific information. The evidence-based movement implies a positivistic epistemological view that influences the information literacy practices. This study focuses how the information literacy practices of hospital librarians in Sweden are constructed and enacted in relation to different epistemological perspectives in healthcare. The analysis is structured around three identified practices of hospital librarians where information work is performed: the clinical practices, the information seeking practices and the health technology assessment (HTA)-practice. In these practices, different epistemological perspectives are present, which affects the information literacy practices of hospital librarians. There is a movement from the holistic knowledge connected to the clinical practices, via specialized knowledge and generic instructions in the information seeking practices, to the most specialized knowledge and positivistic perspective in the HTA-practice.